CONSTRUCTS TRAGETING CD22 AND USES THEREOF

Abstract

Described herein are antibodies (e.g., single chain variable fragment (scFv) antibodies) and constructs comprising antibody moieties that bind to the extracellular domain of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof). Also provided herein are methods of using the same or compositions thereof for the therapeutic treatment of diseases characterized by CD22 expression, in particular, B-cell lymphomas and leukemias.

Claims

1. An anti-CD22 construct comprising an antibody moiety that specifically binds to CD22, wherein the antibody moiety comprises: (a) a light chain variable region (V.sub.L ) comprising a light chain complementarity determining region (LC-CDR) 1, an LC-CDR2, and an LC-CDR3 of the light chain variable region of SEQ ID NO: 218 or 212; and (b) a heavy chain variable region (V.sub.H ) comprising a heavy chain complementarity determining region (HC-CDR) 1, an HC-CDR2, and an HC-CDR3 of the heavy chain variable region of SEQ ID NO: 219 or 213.

2. The anti-CD22 construct of claim 1, wherein the antibody moiety comprises one or more of: the LC-CDR1 having a sequence of HDIRNY (SEQ ID NO: 214), the LC-CDR2 having a sequence of AAS (SEQ ID NO: 215), the LC-CDR3 having a sequence of QQYDGLPLT (SEQ ID NO: 216), the HC-CDR1 having a sequence of GFTFSNYA (SEQ ID NO: 209), the HC-CDR2 having a sequence of ISGSGGST (SEQ ID NO: 210), and the HC-CDR3 having a sequence of ARYGSAAWMDS (SEQ ID NO: 217).

3. The anti-CD22 construct of claim 1 or 2, wherein the light chain variable region has a sequence having at least 90% identity to the sequence of DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTGVPSR FSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKR (SEQ ID NO: 218).

4. The anti-CD22 construct of any one of claims 1 to 3, wherein the heavy chain variable region has a sequence having at least 90% identity to the sequence of QVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVS S (SEQ ID NO: 219).

5. The anti-CD22 construct of claim 1, wherein the antibody moiety comprises one or more of: the LC-CDR1 having a sequence of SSNIGNNY (SEQ ID NO: 206), the LC-CDR2 having a sequence of ENN (SEQ ID NO: 207), the LC-CDR3 having a sequence of GTWDSSLSAGAV (SEQ ID NO: 208), the HC-CDR1 having a sequence of GFTFSNYA (SEQ ID NO: 209), the HC-CDR2 having a sequence of ISGSGGST (SEQ ID NO: 210), and the HC-CDR3 having a sequence of ARPYYDD (SEQ ID NO: 211).

6. The anti-CD22 construct of claim 1 or 5, wherein the light chain variable region has a sequence having at least 90% identity to the sequence of QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLIYENNKRPSGIPD RFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLSAGAVFGGGTKLTVLG (SEQ ID NO: 212).

7. The anti-CD22 construct of any one of claim 1, 5, or 6, wherein the heavy chain variable region has a sequence having at least 90% identity to the sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSAISGSGGSTY YADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARPYYDDWGQGTLVTVSS (SEQ ID NO: 213).

8. The anti-CD22 construct of any one of claims 1 to 7, wherein the construct is a full-length antibody, a Fab, a Fab, a F(ab)2, an Fv, or a single chain Fv (scFv) antibody.

9. The anti-CD22 construct of any one of claims 1 to 8, wherein the construct is a tandem scFv, a diabody (Db), a single chain diabody (scDb), a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, a knob-into-hole (KiH) antibody, a dock and lock (DNL) antibody, a chemically cross-linked antibody, a heteromultimeric antibody, or a heteroconjugate antibody.

10. The anti-CD22 construct of any one of claims 1 to 9, wherein the construct further comprises a second antibody moiety that specifically binds to a second antigen, and wherein the second antigen is an antigen on the surface of a T cell, a natural killer cell, a neutrophil, a monocyte, a macrophage, or a dendritic cell.

11. The anti-CD22 construct of any one of claims 1 to 10, wherein the anti-CD22 construct is a chimeric antigen receptor (CAR).

12. The anti-CD22 construct of claim 11, wherein the CAR comprises an anti-CD22 antibody moiety, a transmembrane domain, and an immune cell signaling domain, wherein the anti-CD22 antibody moiety is a scFv comprising the LC-CDR1, the LC-CDR2, and the LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, the HC-CDR1, the HC-CDR2, and the HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively.

13. The anti-CD22 construct of claim 11, wherein the CAR comprises an anti-CD22 antibody moiety, a transmembrane domain, and an immune cell signaling domain, wherein the anti-CD22 antibody moiety is a scFv comprising the LC-CDR1, the LC-CDR2, and the LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, the HC-CDR1, the HC-CDR2, and the HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively.

14. The anti-CD22 construct of any one of claims 1 to 10, wherein the anti-CD22 construct is a chimeric antibody-T cell receptor (caTCR) comprising an extracellular domain that binds to CD22 and a T cell receptor (TCR) module (TCRM) comprising TCR transmembrane domains.

15. The anti-CD22 construct of claim 14, wherein the caTCR comprises LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, and HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively.

16. The anti-CD22 construct of claim 14, wherein the caTCR comprises LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, and HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively.

17. The anti-CD22 construct of any one of claims 14 to 16, wherein the extracellular domain comprises: (a) a first polypeptide comprising a first antigen-binding region comprising a heavy chain variable region (V.sub.H ) and a C.sub.H1 constant domain; and (b) a second polypeptide chain comprising a second antigen-binding region comprising a light chain variable region (V.sub.L ) and a C.sub.L constant domain, wherein the V.sub.H and the C.sub.H1 constant domain of the first antigen-binding region and the V.sub.L and the C.sub.L constant domain of the second antigen-binding region form a Fab-like antigen-binding module that specifically binds to CD22.

18. The anti-CD22 construct of any one of claims 14 to 17, wherein the extracellular domain further comprises at least one additional antibody moiety that specifically binds to at least one non-CD22 antigen.

19. The anti-CD22 construct of any one of claims 14 to 18, wherein the caTCR is expressed in combination with a chimeric signaling receptor (CSR).

20. The anti-CD22 construct of any one of claims 1 to 10, wherein the anti-CD22 construct is a chimeric signaling receptor (CSR) comprising: (a) an anti-CD22 antibody moiety; (b) a transmembrane module; and (c) a co-stimulatory immune cell signaling module that is capable of providing a co-stimulatory signal to the immune cell, wherein the CSR lacks a functional primary immune cell signaling domain.

21. The anti-CD22 construct of claim 20, wherein the anti-CD22 antibody moiety comprises the LC-CDR1, the LC-CDR2, and the LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, the HC-CDR1, the HC-CDR2, and the HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively.

22. The anti-CD22 construct of claim 20, wherein the anti-CD22 antibody moiety comprises the LC-CDR1, the LC-CDR2, and the LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, the HC-CDR1, the HC-CDR2, and the HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively.

23. The anti-CD22 construct of any one of claims 20 to 22, wherein the CSR is expressed in combination with a caTCR or CAR.

24. A nucleic acid molecule encoding one or more polypeptides contained in the anti-CD22 construct of any one of claims 1 to 23.

25. An expression cassette comprising the nucleic acid molecule of claim 24.

26. A host cell comprising the nucleic acid molecule of claim 24 or the expression cassette of claim 25.

27. A host cell expressing the anti-CD22 construct of any one of claims 1 to 23.

28. A method of preparing an anti-CD22 construct of any one of claims 1 to 23, wherein said method comprising: (a) providing a host cell comprising the nucleic acid molecule of claim 24 or the expression cassette of claim 25, and (b) expressing the nucleic acid molecule or expression cassette in the host cell under conditions that allow for the formation of the anti-CD22 construct.

29. A pharmaceutical composition comprising a therapeutically effective amount of the anti-CD22 construct of any one of claims 1 to 23, the nucleic acid molecule of claim 24, the expression cassette of claim 25, or the host cell of claim 26 or 27, and one or more pharmaceutically acceptable carriers or excipients.

30. A method of treating a B-cell malignancy in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the anti-CD22 construct of any one of claims 1 to 23, the nucleic acid molecule of claim 24, the expression cassette of claim 25, the host cell of claim 26 or 27, or the pharmaceutical composition of claim 29.

31. A method of treating a disease or disorder characterized by CD22 overexpression in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the anti-CD22 construct of any one of claims 1 to 23, the nucleic acid molecule of claim 24, the expression cassette of claim 25, the host cell of claim 26 or 27, or the pharmaceutical composition of claim 29.

32. A method of treatment comprising introducing the nucleic acid molecule of claim 24 or the expression cassette of claim 25 into one or more primary cells isolated from a subject and administering cells comprising the nucleic acid molecule or the expression cassette to the subject.

33. A method of detecting CD22 in a sample, comprising: (a) contacting the sample with the anti-CD22 construct of any one of claims 1 to 23; and (b) detecting the binding, directly or indirectly, between the anti-CD22 construct and any CD22 in the sample.

34. A method of diagnosing a subject suspected of having a CD22-associated disease or disorder, comprising: a) administering an effective amount of the anti-CD22 construct of any one of claims 1 to 23 to the subject; and b) determining the level of the binding, directly or indirectly, between the anti-CD22 construct and any CD22 in the subject, wherein a level of the binding above a threshold level indicates that the subject has the CD22-associated disease or disorder.

35. A method of diagnosing an subject having a B-cell malignancy, comprising: (a) contacting a sample derived from the subject with the anti-CD22 construct of any one of claims 1 to 23; and (b) determining the number of cells bound with the anti-CD22 construct in the sample, wherein a value for the number of cells bound with the anti-CD22 construct above a threshold level indicates that the subject has the B-cell malignancy.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0136] FIGS. 1A-1E show representative MFI of commercial mouse anti-human CD22 antibody (Biolegend, Cat. No. 363505) binding to full-length CD22 only. Full-length CD22 is expressed in Jurkat cells (FIG. 1B) and Raji cells (FIG. 1D).

[0137] FIGS. 2A and 2B show representative MFI of phage clones 1 and 2 binding of human CD22+Raji cells (Raji) and Jurkat cells expressing full-length CD22 (Jurkat-CD22-full) or domains 5-7 of CD22 (Jurkat-CD22 (D5-D7)-GFP) in a flow cytometry assay. CD22- Jurkat cells were used as a negative control.

[0138] FIGS. 3A and 3B show representative MFI of phage clones 1 and 2 binding of NALM-6 cells, which naturally express CD22, in a flow cytometry assay.

[0139] FIGS. 4A-4C illustrate that anti-CD22 CAR (clone 2) displayed specific and potent killing activity against CD22.sup.+ target cells as measured by IFN- release assays.

[0140] FIGS. 5A and 5B illustrate that anti-CD22 CAR (clone 2) demonstrated specific killing against K562+CD22 target cells and no killing against K562 cells.

[0141] FIG. 6 shows that m971 does not compete with anti-CD22 CAR (clone 2) as measured by flow cytometry.

[0142] FIG. 7 shows that caTCR T cells expressing any one of the constructs 2 and 4 and construct combinations 1, 3, 5, and 6 had high killing efficacy in both NALM6 cells and Raji cells.

[0143] FIG. 8 shows that caTCR T cells expressing any one of the constructs 2 and 4 and construct combinations 1, 3, 5, and 6 had high IFN- release levels from NALM6 and Raji cells.

[0144] FIGS. 9A and 9B show that anti-CD22-caTCR T cells expressing any one of the constructs 2 and 4 and construct combinations 3, 5, and 6 maintained high killing efficacy in NALM6 cells up to at least three weeks and after three or more rounds of target engagments. FIG. 9A shows target cell numbers and FIG. 9B shows total cell numbers.

[0145] FIGS. 10A and 10B show that anti-CD22-caTCR T cells expressing any one of the constructs 2 and 4 and construct combinations 1, 3, 5, and 6 had sigfinicant killing efficacy in Raji cells up to about five weeks and after a total of five rounds of target engagements.

[0146] FIG. 11 shows that caTCR T cells expressing any one of the constructs 2, 15-21, and 22 had high killing efficacy in NALM6 cells, Raji cells, and K562 cells.

[0147] FIGS. 12A-12F show that caTCR T cells expressing any one of the constructs 2, 22, 24-27, and 28 had high killing efficacy in NALM6 cells, Raji cells, and K562 cells.

[0148] FIGS. 13A and 13B show that anti-CD22-caTCR T cells expressing any one of the construct combinations 1 and 23 maintained high killing efficacy in NALM6 cells.

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0149] The present invention relates to the discovery of anti-CD22 constructs that have the ability to specifically bind to an extracellular region of CD22. The invention also provides the use of such constructs to treat B-cell malignancies (e.g., CD22.sup.+ B-cell malignancies).

I. CD22

[0150] Cluster of Differentiation 22, or CD22, is a 135-kDa sialic acid-binding cell-surface receptor expressed exclusively during the mature stages of differentiation of the B-cell (Dorken., J. Immunol. 136: 4470, 1986). The predominant form of CD22 in humans is CD22 beta ,which contains seven domains (Wilson et al., J. Exp. Med. 173: 137, 1991). A variant form, CD22 alpha, lacks domains 3 and 4 (Stamenkovic and Seed, Nature 345: 74, 1990). Ligand-binding to human CD22 has been shown to be associated with domains 1 and 2 (also referred to as epitopes 1 and 2) (Engel et al., J. Exp. Med. 181: 1581, 1995). The amino acid sequence of full-length human CD22 is shown in SEQ ID NO: 204 below. SEQ ID NO: 205 further shows the extracellular region containing domains 5-7 of CD22, which anti-CD22 clone 1 and clone 2 bind (see Example 1). Anti-CD22 clone 1 comprises LC-CDR1-3 having the sequences of SEQ ID NOS: 206-208, respectively, and HC-CDR1-3 having the sequences of SEQ ID NOS: 209-211, respectively. Anti-CD22 clone 2 comprises LC-CDR1-3 having the sequences of SEQ ID NOS: 214-216, respectively, and HC-CDR1-3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively.

[0151] SEQ ID NO: 204 (full-length human CD22; underlined: portion of CD22 used in phage display as described in Example 1; bold and underlined: transmembrane domain of CD22):

TABLE-US-00004 MHLLGPWLLLLVLEYLAFSDSSKWVFEHPETLYAWEGACVWIPCTYRALD GDLESFILFHNPEYNKNTSKFDGTRLYESTKDGKVPSEQKRVQFLGDKNK NCTLSIHPV.sub.HLNDSGQLGLRMESKTEKWMERIHLNVSERPFPPHIQLPPE IQESQEVTLTCLLNFSCYGYPIQLQWLLEGVPMRQAAVTSTSLTIKSVFT RSELKFSPQWSHEIGKIVTCQLQDADGKFLSNDTVQLNVKHTPKLEIKVT PSDAIVREGDSVTMTCEVSSSNPEYTTVSWLKDGTSLKKQNTFTLNLREV TKDQSGKYCCQVSNDVGPGRSEEVFLQVQYAPEPSTVQILHSPAVEGSQV EFLCMSLANPLPTNYTWYHNGKEMQGRTEEKV.sub.HIPKILPWHAGTYSCVAE NILGTGQRGPGAELDVQYPPKKVTTVIQNPMPIREGDTVTLSCNYNSSNP SVTRYEWKPHGAWEEPSLGVLKIQNVGWDNTTIACAACNSWCSWASPVAL NVQYAPRDVRVRKIKPLSEIHSGNSVSLQCDFSSSHPKEVQFFWEKNGRL LGKESQLNFDSISPEDAGSYSCWVNNSIGQTASKAWTLEVLYAPRRLRVS MSPGDQVNIEGKSATLTCESDANPPVSHYTWFDWNNQSLPYHSQKLRLEP VKVQHSGAYWCQGTNSVGKGRSPLSTLTVYYSPETIGRRVAVGLGSCLAI LILAICGLKLQRRWKRTQSQQGLQENSSGQSFFVRNKKVRRAPLSEGPHS LGCYNPMMEDGISYTTLRFPEMNIPRTGDAESSEMQRPPPDCDDTVTYSA LHKRQVGDYENVIPDFPEDEGIHYSELIQFGVGERPQAQENVDYVILKH
SEQ ID NO: 205 (extracellular region containing domains 5-7 of CD22; corresponding to amino acids 414-687 of SEQ ID NO: 204):

TABLE-US-00005 DVQYPPKKVTTVIQNPMPIREGDTVTLSCNYNSSNPSVTRYEWKPHGAWE EPSLGVLKIQNVGWDNTTIACAACNSWCSWASPVALNVQYAPRDVRVRKI KPLSEIHSGNSVSLQCDFSSSHPKEVQFFWEKNGRLLGKESQLNFDSISP EDAGSYSCWVNNSIGQTASKAWTLEVLYAPRRLRVSMSPGDQVMEGKSAT LTCESDANPPVSHYTWFDWNNQSLPYHSQKLRLEPVKVQHSGAYWCQGTN SVGKGRSPLSTLTVYYSPETIGRR

[0152] CD22 has been implicated in various cancers, e.g., B-cell related cancers. B-cell related cancers include, but are not limited to, malignant lymphoma (non-Hodgkin's lymphoma), multiple myeloma, and chronic lymphocytic leukemia (CLL, B-cell leukemia (CDS+B lymphocytes)). CD22 expression was recapitulated in human acute lymphoblastic leukemia (ALL)-murine xenografts established with limiting dilution studies, which raises the possibility that CD22 may be expressed on leukemia-initiating cells (Morisot et al., Leukemia 24: 1859, 2010). Furthermore, CD22 expression was found to be maintained in serial studies of 45 patients, including 39 treated with anti-CD22 directed therapy, suggesting that the development of CD22.sup. B-cell precursor acute lymphoblastic leukemia (BCP-ALL) is uncommon (Morisot et al., Leukemia 24: 1859, 2010). The suitability of targeting CD22 in BCP-ALL was tested (Haso et al., Blood 121: 1165, 2013). Lymphoblasts from 111 patients with BCP-ALL were assayed for CD22 expression and all were found to be CD22-positive, with median CD22 expression levels of 3500 sites/cell.

[0153] Moreover, CD22 is highly expressed in over 90% of non-Hodgkin's lymphoma populations (Cesano et al., Blood 100: 350a, 2002). NHLs, a heterogeneous group of cancers principally arising from B lymphocytes, represent approximately 4% of all newly diagnosed cancers (Jemal et al., CA Cancer J Clin, 52: 23, 2002). Aggressive NHL comprises approximately 30-40% of adult NHL (Harris et al., Hematol J. 1: 53, 2001) and includes diffuse large B-cell lymphoma (DLBCL), mantle cell lymphoma (MCL), peripheral T-cell lymphoma, and anaplastic large cell lymphoma. Frontline combination chemotherapy may only treat less than half of the patients with aggressive NHL, and most patients eventually succumb to their disease (Fisher, Oncol. 27 (suppl 12): 2, 2000). There is also evidence that CD22 expression may allow B-cells to become tumorigenic by preventing apoptosis (Otipoby et al., Nature (Loud) 384: 634, 1996). CD22 may function both as a component of the B-cell activation complex (Sato et al., Semin. Immunol. 10: 287, 1998) and as an adhesion molecule (Engel et al., J. Immunol. 150: 4719, 1993). After binding with its natural ligands, CD22 is rapidly internalized, providing a co-stimulatory signal in primary B-cells and proapoptotic signals in neoplastic B-cells (Sato et al., Immunity 5: 551, 1996).

II. Anti-CD22 Constructs and Construct Combinations

[0154] In one aspect, the present invention provides anti-cluster of differentiation-22 (CD22) constructs that comprise an antibody moiety that specifically binds to CD22 (i.e., binds to the sequence of SEQ ID NO: 205 or a portion thereof). In some embodiments, the anti-CD22 construct is an isolated anti-CD22 construct. The specificity of the anti-CD22 construct derives from an anti-CD22 antibody moiety, such as a full-length antibody or antigen-binding fragment thereof, that specifically binds to the CD22 (i.e., binds to the sequence of SEQ ID NO: 205 or a portion thereof). Contemplated anti-CD22 constructs include, for example, full-length anti-CD22 antibodies, multispecific (such as bispecific) anti-CD22 antibodies, anti-CD22 chimeric antigen receptors (CARs), anti-CD22 chimeric antibody-T cell receptors (caTCRs), anti-CD22 chimeric co-stimulatory receptors (CSRs), and anti-CD22 immunoconjugates. An anti-CD22 construct described herein may comprise an antibody moiety that is a full-length anti-CD22 antibody or multispecific (such as bispecific) anti-CD22 antibody. In some embodiemtns, an anti-CD22 construct described herein may be an antibody moiety that is a full-length anti-CD22 antibody or multispecific (such as bispecific) anti-CD22 antibody. In some embodiments, an anti-CD22 construct described herein may be an anti-CD22 chimeric antigen receptors (CAR) or anti-CD22 immunoconjugate.

[0155] In some embodiments, the anti-CD22 constructs can be multispecific (e.g., bispecific). Multispecific anti-CD22 constructs have antibody moieties against more than one target. In some embodiments, a multispecific anti-CD22 constructs has one antibodity moiety against CD22 and one or more additional antibody moieties against one or more non-CD22 antigens (e.g., CD19, CD20, a non-CD22 antigen expressed in B-cell malignancy). In some embodiments, a multispecific anti-CD22 construct can be bispecific, trispecific, quad-specific, or quint-specific. In some embodiments, a multispecific anti-CD22 construct can be a caTCR, a CSR, a CAR, a full-length antibody, a Fab, a Fab, a F(ab)2, an Fv, a single chain Fv (scFv) antibody, a tandem scFv, a diabody (Db), a single chain diabody (scDb), a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, a knob-into-hole (KiH) antibody, a dock and lock (DNL) antibody, a chemically cross-linked antibody, a heteromultimeric antibody, or a heteroconjugate antibody.

[0156] The present invention provides anti-CD22 constructs and methods for using such antibodies for treating CD22-associated diseases. In some embodiments, an anti-CD22 construct described herein recognizes and bind to an epitope within the sequence of SEQ ID NO: 205. An anti-CD22 construct may bind to an epitope comprising at least 7 amino acids (e.g., at least 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, or 40 amino acids) within the sequence of SEQ ID NO: 205. In some embodiments, an anti-CD22 construct may bind to an epitope comprising at least 7 contiguous amino acids (e.g., at least 8, 10, 12, 14, 16, 18, 20, 22, 24, 26, 28, 30, 32, 34, 36, 38, or 40 amino acids) within the sequence of SEQ ID NO: 2. In some embodiments, an anti-CD22 construct described herein binds to the sequence of SEQ ID NO: 205. An anti-CD22 construct described herein may comprise an antibody moiety that is a full-length antibody, a Fab, a Fab, a F(ab)2, an Fv, a single chain Fv (scFv) antibody. In some embodiments, an anti-CD22 construct described herein may be a full-length antibody, a Fab, a Fab, a F(ab)2, an Fv, a single chain Fv (scFv) antibody. In some embodiments, an anti-CD22 construct described herein may be an scFv antibody. In some embodiments, an anti-CD22 construct described herein may comprise an antibody moiety that is a member of an antibody class (i.e., isotype) selected from the group consisting of IgG, IgM, IgA, IgD, IgE, or a fragment thereof

[0157] An anti-CD22 construct may comprise an antibody moiety that specifically binds to CD22, wherein the antibody moiety comprises one or more CDRs, heavy chain variable region, and/or light chain variable region as described herein. In one embodiment, an anti-CD22 construct that specifically binds to CD22 (i.e., binds to the sequence of SEQ ID NO: 205 or a portion thereof) comprises an antibody moiety, wherein the antibody moiety comprises: (1) a light chain variable region comprising a light chain complementarity determining region (LC-CDR) 1, a LC-CDR2, and a LC-CDR3, and (2) a heavy chain variable region comprising a heavy chain complementarity determining region (HC-CDR) 1, an HC-CDR2, and an HC-CDR3, and wherein the antibody moiety comprises one or more of: the LC-CDR1 having the sequence of SSNIGNNY (SEQ ID NO: 206), the LC-CDR2 having the sequence of ENN (SEQ ID NO: 207), the LC-CDR3 having the sequence of GTWDSSLSAGAV (SEQ ID NO: 208), the HC-CDR1 having the sequence of GFTFSNYA (SEQ ID NO: 209), the HC-CDR2 having the sequence of ISGSGGST (SEQ ID NO: 210), and the HC-CDR3 having the sequence of ARPYYDD (SEQ ID NO: 211). In some embodiments, the antibody moiety comprises the sequences of SEQ ID NOS: 206-211.

[0158] In some embodiments, an antibody moiety in an anti-CD22 construct described herein having LC-CDR1 of the sequence of SEQ ID NO: 206, LC-CDR2 of the sequence of SEQ ID NO: 207, and LC-CDR3 of the sequence of SEQ ID NO: 208 may comprise a light chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLIYENNKRPSG IPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDS SLSAGAVFGGGTKLTKLTVLG (SEQ ID NO: 212). In some embodiments, the light chain variable region (e.g., SEQ ID NO: 212) may further be humanized. For example, the light chain variable region may comprise non-human CDRs and framework regions and/or constant regions that are humanized or derived from human antibody sequences. In particular embodiments, the light chain variable region comprises the sequence of SEQ ID NO: 212. In some embodiments, the light chain of the anti-CD22 construct having the LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 206-208, respectively, is a light chain of the lambda isotype.

[0159] In some embodiments, an antibody moiety in an anti-CD22 construct described herein having HC-CDR1 of the sequence of SEQ ID NO: 209, HC-CDR2 of the sequence of SEQ ID NO: 210, and HC-CDR3 of the sequence of SEQ ID NO: 211 may comprise a heavy chain variable region having a sequence that has at least 90% identity (e.g., at least 92%,94%, 96%, 98%, or 99% identity) to the sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSAISGSGG STYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARPYYDDWGQGTLVT VSS (SEQ ID NO: 213). In some embodiments, the heavy chain variable region (e.g., SEQ ID NO: 213) may further be humanized. For example, the heavy chain variable region may comprise non-human CDRs and framework regions and/or constant regions that are humanized or derived from human antibody sequences. In particular embodiments, the heavy chain variable region comprises the sequence of SEQ ID NO: 213.

[0160] In another embodiment, an anti-CD22 construct that specifically binds to CD22 (i.e., binds to the sequence of SEQ ID NO: 205 or a portion thereof) comprises an antibody moiety, wherein the antibody moiety comprises: (1) a light chain variable region comprising a light chain complementarity determining region (LC-CDR) 1, a LC-CDR2, and a LC-CDR3, and (2) a heavy chain variable region comprising a heavy chain complementarity determining region (HC-CDR) 1, an HC-CDR2, and an HC-CDR3, and wherein the antibody moiety comprises one or more of: the LC-CDR1 having the sequence of HDIRNY (SEQ ID NO: 214), the LC-CDR2 having the sequence of AAS (SEQ ID NO: 215), the LC-CDR3 having the sequence of QQYDGLPLT (SEQ ID NO: 216), the HC-CDR1 having the sequence of GFTFSNYA (SEQ ID NO: 209), the HC-CDR2 having the sequence of ISGSGGST (SEQ ID NO: 210), and the HC-CDR3 having the sequence of ARYGSAAWMDS (SEQ ID NO: 217). In some embodiments, the antibody moiety comprises the sequences of SEQ ID NOS: 209, 210, 214-217.

[0161] In some embodiments, an antibody moiety in an anti-CD22 construct described herein having LC-CDR1 of the sequence of SEQ ID NO: 214, LC-CDR2 of the sequence of SEQ ID NO: 215, and LC-CDR3 of the sequence of SEQ ID NO: 216 may comprise a light chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTGV PSRF SGRGSGTDFTLTIS SLQPEDIATYYCQQYDGLPLTFGQGTRLEIKR (SEQ ID NO: 218). In some embodiments, the light chain variable region (e.g., SEQ ID NO: 218) may further be humanized. For example, the light chain variable region may comprise non-human CDRs and framework regions and/or constant regions that are humanized or derived from human antibody sequences. In particular embodiments, the light chain variable region comprises the sequence of SEQ ID NO: 218. In some embodiments, the light chain of the anti-CD22 construct having the LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 214-216, respectively, is a light chain of the lambda isotype.

[0162] In some embodiments, an antibody moiety in an anti-CD22 construct described herein having HC-CDR1 of the sequence of SEQ ID NO: 209, HC-CDR2 of the sequence of SEQ ID NO: 210, and HC-CDR3 of the sequence of SEQ ID NO: 217 may comprise a heavy chain variable region having a sequence that has at least 90% identity (e.g., at least 92%,94%, 96%, 98%, or 99% identity) to the sequence of QVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGG STYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQG TLVTVSS (SEQ ID NO: 219). In some embodiments, the heavy chain variable region (e.g., SEQ ID NO: 219) may further be humanized. For example, the heavy chain variable region may comprise non-human CDRs and framework regions and/or constant regions that are humanized or derived from human antibody sequences. In particular embodiments, the heavy chain variable region comprises the sequence of SEQ ID NO: 219.

[0163] The invention also includes an anti-CD22 construct that comprises an antibody moiety that competes for binding with an antibody moiety having LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, and HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively. The invention also includes an anti-CD22 construct that comprises an antibody moiety that competes for binding with an antibody moiety having a light chain variable region having a sequence of SEQ ID NO: 212 and a heavy chain variable region having a sequence of SEQ ID NO: 213. A skilled artisan in the field has the general knowledge, skills, and methods to identify such competing antibody moieites.

[0164] The invention also includes an anti-CD22 construct that comprises an antibody moiety that competes for binding with an antibody moiety having LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, and HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively. The invention also includes an anti-CD22 construct that comprises an antibody moiety that competes for binding with an antibody moiety having a light chain variable region having a sequence of SEQ ID NO: 218 and a heavy chain variable region having a sequence of SEQ ID NO: 219. A skilled artisan in the field has the general knowledge, skills, and methods to identify such competing antibody moieites.

[0165] Single Chain Variable Fragment (scFv) Antibodies

[0166] An anti-CD22 construct described herein may comprise an antibody moiety that is a single chain Fv (scFv) antibody. In some embodiments, an anti-CD22 construct described herein may be an scFv antibody. An scFv antibody may comprise a light chain variable region and a heavy chain variable region, in which the light chain variable region and the heavy chain variable region may be joined using recombinant methods by a synthetic linker to make a single polypeptide chain. In some embodiments, the scFv may have the structure (N-terminus) light chain variable region-linker-heavy chain variable region (C-terminus), in which the heavy chain variable region is joined to the C-terminus of the light chain variable region by way of a linker. In other embodiments, the scFv may have the structure (N-terminus) heavy chain variable region-linker-light chain variable region (C-terminus), in which the light chain variable region is joined to the C-terminus of the heavy chain variable region by way of a linker. A linker may be a polypeptide including 2 to 200 (e.g., 2, 3, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, or 200) amino acids. Suitable linkers may contain flexible amino acid residues such as glycine and serine. In certain embodiments, a linker may contain motifs, e.g., multiple or repeating motifs, of GS, GGS, GGGGS (SEQ ID NO: 220), GGSG (SEQ ID NO: 221), or SGGG (SEQ ID NO: 222). In some embodiments, a linker may have the sequence GSGS (SEQ ID NO: 223), GSGSGS (SEQ ID NO: 224), GSGSGSGS (SEQ ID NO: 225), GSGSGSGSGS (SEQ ID NO: 226), GGSGGS (SEQ ID NO: 227), GGSGGSGGS (SEQ ID NO: 228), GGSGGSGGSGGS (SEQ ID NO: 229). GGSG (SEQ ID NO: 230), GGSGGGSG (SEQ ID NO: 231), or GGSGGGSGGGSG (SEQ ID NO: 232). In other embodiments, a linker may also contain amino acids other than glycine and serine, e.g., SRGGGGSGGGGSGGGGSLEMA (SEQ ID NO: 233).

[0167] Purification Tag

[0168] In some embodiments, a purification tag may be joined to an anti-CD22 construct described herein (e.g., an anti-CD22 scFv). A purification tag refers to a peptide of any length that can be used for purification, isolation, or identification of a polypeptide. A purification tag may be joined to a polypeptide (e.g., joined to the N- or C-terminus of the polypeptide) to aid in purifying the polypeptide and/or isolating the polypeptide from, e.g., a cell lysate mixture. In some embodiments, the purification tag binds to another moiety that has a specific affinity for the purification tag. In some embodiments, such moieties which specifically bind to the purification tag are attached to a solid support, such as a matrix, a resin, or agarose beads. Examples of a purification tag that may be joined to an anti-CD22 construct (e.g., an anti-CD22 scFv) include, but are not limited to, a hexa-histidine peptide, a hemagglutinin (HA) peptide, a FLAG peptide, and a myc peptide. In some embodiments, two or more purification tag may be joined to a construct, e.g., a hexa-histidine peptide and a HA peptide. A hexa-histidine peptide (HHHHHH (SEQ ID NO: 234)) binds to nickel-functionalized agarose affinity column with micromolar affinity. In some embodiments, an HA peptide includes the sequence YPYDVPDYA (SEQ ID NO: 235) or YPYDVPDYAS (SEQ ID NO: 236). In some embodiments, an HA peptide includes integer multiples of the sequence YPYDVPDYA (SEQ ID NO: 235) or YPYDVPDYAS (SEQ ID NO: 236) in tandem series, e.g., 3YPYDVPDYA or 3YPYDVPDYAS. In some embodiments, a FLAG peptide includes the sequence DYKDDDDK (SEQ ID NO: 237). In some embodiments, a FLAG peptide includes integer multiples of the sequence DYKDDDDK (SEQ ID NO: 237) in tandem series, e.g., 3DYKDDDDK. In some embodiments, a myc peptide includes the sequence EQKLISEEDL (SEQ ID NO: 238). In some embodiments, a myc peptide includes integer multiples of the sequence EQKLISEEDL in tandem series, e.g., 3EQKLISEEDL.

[0169] Anti-CD22 scFv Antibodies

[0170] In some embodiments, an anti-CD22 construct having the LC-CDR1, LC-CDR2, LC-CDR3, HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 206-211, respectively, may be an scFv antibody. In some embodiments, the heavy chain variable region having at least 90% identity (e.g., at least 92%,94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 213 may be joined to the N- or C-terminus of the light chain variable region having at least 90% identity (e.g., at least 92%,94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 212 by way of a linker. In particular embodiments, the anti-CD22 scFv antibody has the sequence of SEQ ID NO: 239 shown below. SEQ ID NO: 239 (anti-CD22 scFv antibody; plain text (no bold or underline): light chain variable region and heavy chain variable region; bold: linker; and underlined: purification tags (hexa-histidine tag and HA tag)

TABLE-US-00006 QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLIY ENNKRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLSAGA VFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVESGGGLVQPGGS LRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGR FTISRDNSKNTLYLQMNSLRAEDTAVYYCARPYYDDWGQGTLVTVSSTSG QAGQHHHHHHGAYPYDVPDYAS

[0171] In some embodiments, an anti-CD22 construct having the LC-CDR1, LC-CDR2, LC-CDR3, HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 214-216, 209, 210, and 217, respectively, may be an scFv antibody. In some embodiments, the heavy chain variable region having at least 90% identity (e.g., at least 92%,94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 219 may be joined to the N- or C-terminus of the light chain variable region having at least 90% identity (e.g., at least 92%,94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 218 by way of a linker. In particular embodiments, the anti-CD22 scFv antibody has the sequence of SEQ ID NO: 240 shown below. SEQ ID NO: 240 (anti-CD22 scFv antibody; plain text (no bold or underline): light chain variable region and heavy chain variable region; bold: linker; and underlined: purification tags (hexa-histidine tag and HA tag)

TABLE-US-00007 DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYA ASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQ GTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLS CAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTIS RDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSTSG QAGQHHHHHHGAYPYDVPDYAS

[0172] Multispecific Antibodies

[0173] Anti-CD22 constructs described herein may comprise an antibody moiety that is a multispecific antibody. In some embodiments, an anti-CD22 construct described herein may be a multispecific antibody. A multispecific antibody may comprise an anti-CD22 binding moiety and a second binding moiety (such as a second antigen-binding moiety). Multispecific antibodies are antibodies that have binding specificities for at least two different antigens or epitopes (e.g., bispecific antibodies have binding specificities for two antigens or epitopes). Multispecific antibodies with more than two specificities are also contemplated. For example, trispecific antibodies can be prepared (see, e.g., Tutt et al. J. Immunol. 147: 60 (1991)). It is to be appreciated that one of skill in the art could select appropriate features of individual multispecific antibodies described herein to combine with one another to form a multispecific antibodies of the invention.

[0174] Thus, for example, in some embodiments, there is provided a multispecific (e.g., bispecific) anti-CD22 antibody comprising a) an anti-CD22 binding moiety that specifically binds to an extracellular region of CD22 (i.e., binds to the sequence of SEQ ID NO: 205 or a portion thereof), and b) a second binding moiety (such as an antigen-binding moiety). In some embodiments, the second binding moiety specifically binds to a different antigen. In some embodiments, the second binding moiety specifically binds to an antigen on the surface of a cell, such as a cytotoxic cell. In some embodiments, the second binding moiety specifically binds to an antigen on the surface of a lymphocyte, such as a T cell, an NK cell, a neutrophil, a monocyte, a macrophage, or a dendritic cell. In some embodiments, the second binding moiety specifically binds to an effector T cell, such as a cytotoxic T cell (also known as cytotoxic T lymphocyte (CTL) or T killer cell).

[0175] In some embodiments, the second binding moiety specifically binds to a tumor antigen. Examples of tumor antigens include, but are not limited to, alpha fetoprotein (AFP), CA15-3, CA27-29, CA19-9, CA-125, calretinin, carcinoembryonic antigen, CD34, CD99, CD117, chromogranin, cytokeratin, desmin, epithelial membrane protein (EMA), Factor VIII, CD31 FL1, glial fibrillary acidic protein (GFAP), gross cystic disease fluid protein (GCDFP-15), HMB-45, human chorionic gonadotropin (hCG), inhibin, keratin, CD45, a lymphocyte marker, MART-1 (Melan-A), Myo Dl, muscle-specific actin (MSA), neurofilament, neuron-specific enolase (NSE), placental alkaline phosphatase (PLAP), prostate-specific antigen, S100 protein, smooth muscle actin (SMA), synaptophysin, thyroglobulin, thyroid transcription factor- 1, tumor M2-PK, and vimentin.

[0176] In some embodiments, the second antigen-binding moiety in a bispecific antibody binds to CD3. In some embodiments, the second antigen-binding moiety specifically binds to CD3. In some embodiments, the second antigen-binding moiety specifically binds to an agonistic epitope of CD3. The term agonistic epitope, as used herein, means (a) an epitope that, upon binding of the multispecific antibody, optionally upon binding of several multispecific antibodies on the same cell, allows said multispecific antibodies to activate T cell receptor (TCR) signaling and induce T cell activation, and/or (b) an epitope that is solely composed of amino acid residues of the epsilon chain of CD3 and is accessible for binding by the multispecific antibody, when presented in its natural context on T cells (i.e. surrounded by the TCR, the CD3 chain, etc.), and/or (c) an epitope that, upon binding of the multispecific antibody, does not lead to stabilization of the spatial position of CD3relative to CD3.

[0177] In some embodiments, the second antigen-binding moiety binds specifically to an antigen on the surface of an effector cell, including for example CD3, CD3, CD3, CD3, CD28, CD16a, CD56, CD68, GDS2D, CD3, CD3, CD3, CD3, CD28, CD16a, CD56, CD68, GDS2D, OX40, GITR, CD137, CD27, CD4OL and HVEM. In other embodiments, the second antigen-binding moiety binds to a component of the complement system, such as Cq. Cq is a subunit of the C1 enzyme complex that activates the serum complement system. In other embodiments, the second antigen-binding moiety specifically binds to an Fc receptor. In some embodiments, the second antigen-binding moiety specifically binds to an Fc receptor (FcR). The FcR may be an FcRIII present on the surface of natural killer (NK) cells or one of FcRI, FcRIIA, FcRIIBI, FcRIIB2, and FcRIIIB present on the surface of macrophages, monocytes, neutrophils and/or dendritic cells. In some embodiments, the second antigen-binding moiety is an Fc region or functional fragment thereof. A functional fragment as used in this context refers to a fragment of an antibody Fc region that is still capable of binding to an FcR, in particular to an FcR, with sufficient specificity and affinity to allow an FcR bearing effector cell, in particular a macrophage, a monocyte, a neutrophil and/or a dendritic cell, to kill the target cell by cytotoxic lysis or phagocytosis. A functional Fc fragment is capable of competitively inhibiting the binding of the original, full-length Fc portion to an FcR such as the activating FcRI. In some embodiments, a functional Fc fragment retains at least 30%, 40%, 50%, 60%, 70%, 80%, 90% or 95% of its affinity to an activating FcR. In some embodiments, the Fc region or functional fragment thereof is an enhanced Fc region or functional fragment thereof. The term enhanced Fc region, as used herein, refers to an Fc region that is modified to enhance Fc receptor-mediated effector-functions, in particular antibody-dependent cell-mediated cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and antibody-mediated phagocytosis. This can be achieved as known in the art, for example by altering the Fc region in a way that leads to an increased affinity for an activating receptor (e.g. FcRIIIA (CD16A) expressed on natural killer (NK) cells) and/or a decreased binding to an inhibitory receptor (e.g. FcRIIB1/B2 (CD32B)).

[0178] In some embodiments, the multispecific anti-CD22 antibodies allow killing of CD22-presenting target cells and/or can effectively redirect CTLs to lyse CD22-presenting target cells. In some embodiments, the multispecific (e.g., bispecific) anti-CD22 antibodies of the present invention show an in vitro EC50 ranging from 10 to 500 ng/ml, and is able to induce redirected lysis of about 50% of the target cells through CTLs at a ratio of CTLs to target cells of from about 1:1 to about 50:1 (such as from about 1:1 to about 15:1, or from about 2:1 to about 10:1).

[0179] In some embodiments, the multispecific (e.g., bispecific) anti-CD22 antibody is capable of cross-linking a stimulated or unstimulated CTL and the target cell in such a way that the target cell is lysed. This offers the advantage that no generation of target-specific T cell clones or common antigen presentation by dendritic cells is required for the multispecific anti-CD22 antibody to exert its desired activity. In some embodiments, the multispecific anti-CD22 antibody of the present invention is capable of redirecting CTLs to lyse the target cells in the absence of other activating signals. In some embodiments, the second antigen-binding moiety specifically binds to CD3 (e.g., specifically binds to CDR), and signaling through CD28 and/or IL-2 is not required for redirecting CTLs to lyse the target cells.

[0180] Methods for measuring the preference of the multispecific anti-CD22 antibody to simultaneously bind to two antigens (e.g., antigens on two different cells) are within the normal capabilities of a person skilled in the art. For example, when the second binding moiety specifically binds to CD3, the multispecific anti-CD22 antibody may be contacted with a mixture of CD3+/CD22- cells and CD3-/CD22.sup.+ cells. The number of multispecific anti-CD22 antibody-positive single cells and the number of cells cross-linked by multispecific anti-CD22 antibodies may then be assessed by microscopy or fluorescence-activated cell sorting (FACS) as known in the art.

[0181] A multispecific anti-CD22 antibody may comprise a) an anti-CD22 binding moiety comprising i) a light chain variable region comprising LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, and ii) a heavy chain variable region comprising HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively, and b) a second antigen-binding moiety. In another embodiment, a multispecific anti-CD22 antibody may comprise a) an anti-CD22 binding moiety comprising i) a light chain variable region comprising LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, and ii) a heavy chain variable region comprising HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively, and b) a second antigen-binding moiety.

[0182] In some embodiments, the multispecific anti-CD22 antibody is, for example, a diabody (Db), a single-chain diabody (scDb), a tandem scDb (Tandab), a linear dimeric scDb (LD-scDb), a circular dimeric scDb (CD-scDb), a di-diabody, a tandem scFv, a tandem di-scFv (e.g., a bispecific T cell engager), a tandem tri-scFv, a tri(a)body, a bispecific Fab2, a di-miniantibody, a tetrabody, an scFv-Fc-scFv fusion, a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, an IgG-scFab, an scFab-ds-scFv, an Fv2-Fc, an IgG-scFv fusion, a dock and lock (DNL) antibody, a knob-into-hole (KiH) antibody (bispecific IgG prepared by the KiH technology), a DuoBody (bispecific IgG prepared by the Duobody technology), a heteromultimeric antibody, or a heteroconjugate antibody. In some embodiments, the multispecific anti-CD22 antibody is a tandem scFv (e.g., a tandem di-scFv, such as a bispecific T cell engager).

[0183] Tandem scFv

[0184] The multispecific anti-CD22 antibody in some embodiments is a tandem scFv comprising a first scFv comprising an anti-CD22 binding moiety and a second scFv (also referred to herein as a tandem scFv multispecific anti-CD22 antibody). In some embodiments, the tandem scFv multispecific anti-CD22 antibody further comprises at least one (such as at least about any of 2, 3, 4, 5, or more) additional scFv.

[0185] In some embodiments, there is provided a tandem scFv multispecific (e.g., bispecific) anti-CD22 antibody comprising a) a first scFv that specifically binds to an extracellular region of CD22, and b) a second scFv. In some embodiments, the first scFv comprises a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 206-208, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 212 and a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209-211, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 213, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker (e.g., SRGGGGSGGGGSGGGGSLEMA (SEQ ID NO: 233)). In some embodiments, the first scFv comprises the light chain variable region sequence of SEQ ID NO: 212 and the heavy chain variable region sequence of SEQ ID NO: 213 joined to each other via a linker (e.g., SRGGGGSGGGGSGGGGSLEMA (SEQ ID NO: 233)). The first scFv may bind to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof).

[0186] In some embodiments, there is provided a tandem scFv multispecific (e.g., bispecific) anti-CD22 antibody comprising a) a first scFv that specifically binds to an extracellular region of CD22, and b) a second scFv. In some embodiments, the first scFv comprises a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 214-216, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 218 and a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209, 210, and 217, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 219, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker (e.g., SRGGGGSGGGGSGGGGSLEMA (SEQ ID NO: 233)). In some embodiments, the first scFv comprises the light chain variable region sequence of SEQ ID NO: 218 and the heavy chain variable region sequence of SEQ ID NO: 219 joined to each other via a linker (e.g., SRGGGGSGGGGSGGGGSLEMA (SEQ ID NO: 233)). The first scFv may bind to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof).

[0187] In some embodiments, the second scFv specifically binds to another antigen. In some embodiments, the second scFv specifically binds to an antigen on the surface of a cancer cell, such as a CD22-presenting cell. In some embodiments, the second scFv specifically binds to an antigen on the surface of a cell that does not express CD22. In some embodiments, the second scFv specifically binds to an antigen on the surface of a cytotoxic cell. In some embodiments, the second scFv specifically binds to an antigen on the surface of a lymphocyte, such as a T cell, an NK cell, a neutrophil, a monocyte, a macrophage, or a dendritic cell. In some embodiments, the second scFv specifically binds to an antigen on the surface of an effector T cell, such as a cytotoxic T cell. In some embodiments, the second scFv specifically binds to an antigen on the surface of an effector cell, including for example CD3, CD3, CD3, CD3, CD28, CD16a, CD56, CD68, GDS2D, OX40, GITR, CD137, CD27, CD4OL and HVEM.

[0188] In some embodiments, the first scFv is human, humanized, or semi-synthetic. In some embodiments, the second scFv is human, humanized, or semi-synthetic. In some embodiments, both the first scFv and the second scFv are human, humanized, or semi-synthetic. In some embodiments, the tandem scFv multispecific anti-CD22 antibody further comprises at least one (such as at least about any of 2, 3, 4, 5, or more) additional scFv.

[0189] In some embodiments, there is provided a tandem scFv multispecific (e.g., bispecific) anti-CD22 antibody comprising a) a first scFv that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), and b) a second scFv, wherein the tandem scFv multispecific anti-CD22 antibody is a tandem di-scFv or a tandem tri-scFv. In some embodiments, the tandem scFv multispecific anti-CD22 antibody is a tandem di-scFv. In some embodiments, the tandem scFv multispecific anti-CD22 antibody is a bispecific T-cell engager.

[0190] In some embodiments, the tandem di-scFv bispecific anti-CD22 antibody binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof) with a Kd between about 0.1 pM to about 500 nM (such as about any of 0.1 pM, 1.0 pM, 10 pM, 50 pM, 100 pM, 500 pM, 1 nM, 10 nM, 50 nM, 100 nM, or 500 nM, including any ranges between these values). In some embodiments, the tandem di-scFv bispecific anti-CD22 antibody binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof) with a Kd between about 1 nM to about 500 nM (such as about any of 1, 10, 25, 50, 75, 100, 150, 200, 250, 300, 350, 400, 450, or 500 nM, including any ranges between these values).

[0191] A variety of technologies are known in the art for designing, constructing, and/or producing multispecific antibodies. Multispecific antibodies may be constructed that either utilize the full immunoglobulin framework (e.g., IgG), single chain variable fragment (scFv), or combinations thereof. Bispecific antibodies may be composed of two scFv units in tandem as described above. In the case of anti-tumor immunotherapy, bispecific antibodies that comprise two single chain variable fragments (scFvs) in tandem may be designed such that an scFv that binds a tumor antigen is linked with an scFv that engages T cells, i.e., by binding CD3 on the T cells. Thus, T cells are recruited to a tumor site to mediate killing of the tumor cells. Bispecific antibodies can be made, for example, by combining heavy chains and/or light chains that recognize different epitopes of the same or different antigen. In some embodiments, by molecular function, a bispecific binding agent binds one antigen (or epitope) on one of its two binding arms (one V.sub.H/V.sub.L pair), and binds a different antigen (or epitope) on its second arm (a different V.sub.H/V.sub.L pair). By this definition, a bispecific binding agent has two distinct antigen binding arms (in both specificity and CDR sequences), and is monovalent for each antigen to which it binds. In certain embodiments, a bispecific binding agent according to the present invention comprises a first and a second scFv. In some certain embodiments, a first scFv is linked to the C-terminal end of a second scFv. In some certain embodiments, a second scFv is linked to the C-terminal end of a first scFv. In some certain embodiments, scFvs are linked to each other via a linker (e.g., SRGGGGSGGGGSGGGGSLEMA (SEQ ID NO: 233)). In some certain embodiments, scFvs are linked to each other without a linker.

[0192] Chimeric Antigen Receptor (CAR) Constructs

[0193] In some embodiments, an anti-CD22 construct may be an anti-CD22 CAR. In some embodiments, the anti-CD22 CAR comprises a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (SEQ ID NO: 205 or a portion thereof) and b) an intracellular signaling domain. A transmembrane domain may be present between the extracellular domain and the intracellular domain.

[0194] Between the extracellular domain and the transmembrane domain of the anti-CD22 CAR, or between the intracellular domain and the transmembrane domain of the anti-CD22 CAR, there may be a spacer domain. The spacer domain can be any oligo- or polypeptide that functions to link the transmembrane domain to the extracellular domain or the intracellular domain in the polypeptide chain. A spacer domain may comprise up to about 300 amino acids, including for example about 10 to about 100, or about 25 to about 50 amino acids.

[0195] The transmembrane domain may be derived either from a natural or from a synthetic source. Where the source is natural, the domain may be derived from any membrane-bound or transmembrane protein. Transmembrane regions of particular use in this invention may be derived from (i.e. comprise at least the transmembrane region(s) of) the , , , , or chain of the T-cell receptor, CD28, CD3, CD3, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154. In some embodiments, the transmembrane domain may be synthetic, in which case it may comprise predominantly hydrophobic residues such as leucine and valine. In some embodiments, a triplet of phenylalanine, tryptophan, and valine may be found at each end of a synthetic transmembrane domain. In some embodiments, a short oligo- or polypeptide linker, having a length of, for example, between about 2 and about 10 (such as about any of 2, 3, 4, 5, 6, 7, 8, 9, or 10) amino acids in length may form the linkage between the transmembrane domain and the intracellular signaling domain of the anti-CD22 CAR. In some embodiments, the linker is a glycine-serine doublet.

[0196] In some embodiments, the transmembrane domain that naturally is associated with one of the sequences in the intracellular domain of the anti-CD22 CAR is used (e.g., if an anti-CD22 CAR intracellular domain comprises a CD28 co-stimulatory sequence, the transmembrane domain of the anti-CD22 CAR is derived from the CD28 transmembrane domain). In some embodiments, the transmembrane domain can be selected or modified by amino acid substitution to avoid binding of such domains to the transmembrane domains of the same or different surface membrane proteins to minimize interactions with other members of the receptor complex.

[0197] The intracellular signaling domain of the anti-CD22 CAR is responsible for activation of at least one of the normal effector functions of the immune cell in which the anti-CD22 CAR has been placed in. Effector function of a T cell, for example, may be cytolytic activity or helper activity including the secretion of cytokines. Thus the term intracellular signaling domain refers to the portion of a protein which transduces the effector function signal and directs the cell to perform a specialized function. While usually the entire intracellular signaling domain can be employed, in many cases it is not necessary to use the entire chain. To the extent that a truncated portion of the intracellular signaling domain is used, such truncated portion may be used in place of the intact chain as long as it transduces the effector function signal. The term intracellular signaling sequence is thus meant to include any truncated portion of the intracellular signaling domain sufficient to transduce the effector function signal.

[0198] Examples of intracellular signaling domains for use in the anti-CD22 CAR of the invention include the cytoplasmic sequences of the T cell receptor (TCR) and co-receptors that act in concert to initiate signal transduction following antigen receptor engagement, as well as any derivative or variant of these sequences and any synthetic sequence that has the same functional capability.

[0199] It is known that signals generated through the TCR alone are insufficient for full activation of the T cell and that a secondary or co-stimulatory signal is also required. Thus, T cell activation can be said to be mediated by two distinct classes of intracellular signaling sequence: those that initiate antigen-dependent primary activation through the TCR (primary signaling sequences) and those that act in an antigen-independent manner to provide a secondary or co-stimulatory signal (co-stimulatory signaling sequences).

[0200] Primary signaling sequences regulate primary activation of the TCR complex either in a stimulatory way, or in an inhibitory way. Primary signaling sequences that act in a stimulatory manner may contain signaling motifs which are known as immunoreceptor tyrosine-based activation motifs or ITAMs. The anti-CD22 CAR constructs in some embodiments comprise one or more ITAMs.

[0201] Examples of ITAM containing primary signaling sequences that are of particular use in the invention include those derived from TCR, FcR, FcR, CD3, CD3, CD3, CD5, CD22, CD79a, CD79b, and CD66d.

[0202] In some embodiments, the anti-CD22 CAR comprises a primary signaling sequence derived from CD3. For example, the intracellular signaling domain of the CAR can comprise the CD3 intracellular signaling sequence by itself or combined with any other desired intracellular signaling sequence(s) useful in the context of the anti-CD22 CAR of the invention. For example, the intracellular domain of the anti-CD22 CAR can comprise a CD3 intracellular signaling sequence and a co-stimulatory signaling sequence. The co-stimulatory signaling sequence can be a portion of the intracellular domain of a co-stimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like.

[0203] In some embodiments, the intracellular signaling domain of the anti-CD22 CAR comprises the intracellular signaling sequence of CD3 and the intracellular signaling sequence of CD28. In some embodiments, the intracellular signaling domain of the anti-CD22 CAR comprises the intracellular signaling sequence of CD3 and the intracellular signaling sequence of 4-1BB. In some embodiments, the intracellular signaling domain of the anti-CD22 CAR comprises the intracellular signaling sequence of CD3 and the intracellular signaling sequences of CD28 and 4-1BB.

[0204] Thus, for example, in some embodiments, there is provided an anti-CD22 CAR comprising a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), b) a transmembrane domain, and c) an intracellular signaling domain. In some embodiments, the intracellular signaling domain is capable of activating an immune cell. In some embodiments, the intracellular signaling domain comprises a primary signaling sequence and a co-stimulatory signaling sequence. In some embodiments, the primary signaling sequence comprises a CD3 intracellular signaling sequence. In some embodiments, the co-stimulatory signaling sequence comprises a CD28 and/or 4-1BB intracellular signaling sequence. In some embodiments, the intracellular domain comprises a CD3 intracellular signaling sequence and a CD28 and/or 4-1BB intracellular signaling sequence.

[0205] In some embodiments, the anti-CD22 CAR comprises an anti-CD22 antibody moiety comprising i) a light chain variable region comprising LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, and ii) a heavy chain variable region comprising HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively. In other embodiments, the anti-CD22 CAR comprises an anti-CD22 antibody moiety comprising i) a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 206-208, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 212, and ii) a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209-211, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 213, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker.

[0206] In some embodiments, the anti-CD22 CAR comprises an anti-CD22 antibody moiety comprising i) a light chain variable region comprising LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, and ii) a heavy chain variable region comprising HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively. In other embodiments, the anti-CD22 CAR comprises an anti-CD22 antibody moiety comprising i) a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 214-216, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 218, and ii) a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209, 210, and 217, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 219, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker.

[0207] Chimeric Antibody-T Cell Receptor (caTCR) Constructs

[0208] In some embodiments, the anti-CD22 construct is a chimeric antibody-T cell receptor construct (referred to herein as caTCR), and the anti-CD22 chimeric antibody-T cell receptor is an anti-CD22 caTCR. In some embodiments, the anti-CD22 caTCR comprises a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof) and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module.

[0209] In some embodiments, the anti-CD22 caTCR comprises a first polypeptide chain and a second polypeptide chain. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g., peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the anti-CD22 caTCR is a heterodimer comprising the first polypeptide chain and the second polypeptide chain. In some embodiments, the first polypeptide chain and the second polypeptide chain are linked by at least one disulfide bond. The specificity of the anti-CD22 caTCR derives from an antibody moiety that confers binding specificity to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof). In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv. The capability of the anti-CD22 caTCR to recruit a TCR-associated signaling module derives from a T cell receptor module (TCRM). In some embodiments, the TCRM comprises the transmembrane module of a TCR (such as an TCR or a TCR). In some embodiments, the TCRM further comprises one or both of the connecting peptides or fragments thereof of a TCR. In some embodiments, the anti-CD22 caTCR further comprises at least one intracellular domain. In some embodiments, one or more of the at least one intracellular domain of the anti-CD22 caTCR comprises a sequence from the intracellular domain of a TCR. In some embodiments, the antibody moiety is contained in an extracellular domain of the anti-CD22 caTCR. In some embodiments, the anti-CD22 caTCR further comprises one or more peptide linkers between the antibody moiety and the TCRM to optimize the length of the extracellular domain.

[0210] In some embodiments, the antibody moiety is a Fab-like antigen-binding module that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), comprising a) a first polypeptide chain comprising a first antigen-binding region comprising a V.sub.H antibody domain and a C.sub.H1 antibody domain and b) a second polypeptide chain comprising a second antigen-binding region comprising a V.sub.L antibody domain and a C.sub.L antibody domain. In some embodiments, the first antigen-binding region comprises the V.sub.H antibody domain amino-terminal to the C.sub.H1 antibody domain and/or the second antigen-binding region comprises the V.sub.L antibody domain amino-terminal to the C.sub.L antibody domain. In some embodiments, there is a peptide linker between the V.sub.L and C.sub.L antibody domains and/or a peptide linker between the V.sub.H and C.sub.H1 antibody domains. In some embodiments, all of the V.sub.L antibody domain and V.sub.H antibody domain CDRs are derived from the same antibody moiety. In some embodiments, the V.sub.L antibody domain and the V.sub.H antibody domain comprise antibody CDRs derived from more than one antibody moiety. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g., peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the first and second antigen-binding regions are linked by a disulfide bond. In some embodiments, the first and second antigen-binding regions are linked by a disulfide bond between a residue in the C.sub.H1 domain and a residue in the C.sub.L domain. In some embodiments, the C.sub.H1 domain is derived from an IgG (e.g, IgG1, IgG2, IgG3, or IgG4) heavy chain, optionally human. In some embodiments, the C.sub.H1 domain is a variant comprising one or more modifications (e.g., amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the CL domain is derived from a light chain of the kappa or lambda isotype. In some embodiments, the C.sub.L domain is a variant comprising one or more modifications (e.g., amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the C.sub.H1 and/or C.sub.L domains comprise one or more modifications that do not substantially alter their binding affinities for one another. In some embodiments, the C.sub.H1 and/or C.sub.L domains comprise one or more modifications that increase their binding affinities for one another and/or introduce a non-naturally occurring disulfide bond. In some embodiments, the Fab-like antigen-binding module is human, humanized, chimeric, semi-synthetic, or fully synthetic.

[0211] In some embodiments, the antibody moiety is a Fab-like antigen-binding module that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), comprising a) a first polypeptide chain comprising a first antigen-binding region comprising a V.sub.L antibody domain and a C.sub.H1 antibody domain and b) a second polypeptide chain comprising a second antigen-binding region comprising a V.sub.H antibody domain and a C.sub.L antibody domain. In some embodiments, the first antigen-binding region comprises the V.sub.L antibody domain amino-terminal to the C.sub.H1 antibody domain and/or the second antigen-binding region comprises the V.sub.H antibody domain amino-terminal to the C.sub.L antibody domain. In some embodiments, there is a peptide linker between the V.sub.H and C.sub.L antibody domains and/or a peptide linker between the V.sub.L and C.sub.H1 antibody domains. In some embodiments, all of the V.sub.L antibody domain and V.sub.H antibody domain CDRs are derived from the same antibody moiety. In some embodiments, the V.sub.L antibody domain and the V.sub.H antibody domain comprise antibody CDRs derived from more than one antibody moiety. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g., peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the first and second antigen-binding regions are linked by a disulfide bond. In some embodiments, the first and second antigen-binding regions are linked by a disulfide bond between a residue in the C.sub.H1 domain and a residue in the C.sub.L domain. In some embodiments, the C.sub.H1 domain is derived from an IgG (e.g, IgG1, IgG2, IgG3, or IgG4) heavy chain, optionally human. In some embodiments, the C.sub.H1 domain is a variant comprising one or more modifications (e.g., amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the CL domain is derived from a light chain of the kappa or lambda isotype. In some embodiments, the C.sub.L domain is a variant comprising one or more modifications (e.g., amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the C.sub.H1 and/or C.sub.L domains comprise one or more modifications that do not substantially alter their binding affinities for one another. In some embodiments, the C.sub.H1 and/or C.sub.L domains comprise one or more modifications that increase their binding affinities for one another and/or introduce a non-naturally occurring disulfide bond. In some embodiments, the Fab-like antigen-binding module is human, humanized, chimeric, semi-synthetic, or fully synthetic.

[0212] In some embodiments, the antibody moiety is an Fv-like antigen-binding module that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), comprising a) a first polypeptide chain comprising a first antigen-binding region comprising a V.sub.H antibody domain and optionally a first TCR constant domain from a T cell receptor subunit; and b) a second polypeptide chain comprising a second antigen-binding region comprising a V.sub.L antibody domain and optionally a second TCR constant domain from a T cell receptor subunit. In some embodiments, the first antigen-binding region comprises the V.sub.H antibody domain amino-terminal to the first TCR constant domain and/or the second antigen-binding region comprises the V.sub.L antibody domain amino-terminal to the second TCR constant domain. In some embodiments, there is a peptide linker between the V.sub.L antibody domain and the first TCR constant domain and/or a peptide linker between the V.sub.H antibody domain and the second TCR constant domain. In some embodiments, all of the V.sub.L antibody domain and V.sub.H antibody domain CDRs are derived from the same antibody moiety. In some embodiments, the V.sub.L antibody domain and the V.sub.H antibody domain comprise antibody CDRs derived from more than one antibody moiety. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g., peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the first and second antigen-binding regions are linked by a disulfide bond. In some embodiments, the first and second antigen-binding regions are linked by a disulfide bond between a residue in the first TCR constant domain and a residue in the second TCR constant domain. In some embodiments, the first TCR constant domain is derived from a TCR subunit, optionally human, and/or the second TCR constant domain is derived from a TCR subunit, optionally human. In some embodiments, the first TCR constant domain is derived from a TCR subunit, optionally human, and/or the second TCR constant domain is derived from a TCR subunit, optionally human. In some embodiments, the first and/or second TCR constant domain is a variant comprising one or more modifications (e.g., amino acid substitutions, insertions, and/or deletions) compared to the sequence from which it is derived. In some embodiments, the first and/or second TCR constant domains comprise one or more modifications that do not substantially alter their binding affinities for one another. In some embodiments, the first and/or second TCR constant domains comprise one or more modifications that increase their binding affinities for one another and/or introduce a non-naturally occurring disulfide bond. In some embodiments, the Fv-like antigen-binding module is human, humanized, chimeric, semi-synthetic, or fully synthetic.

[0213] In some embodiments, the antibody moiety is an scFv that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), comprising a) a polypeptide chain comprising a V.sub.H antibody domain and a V.sub.L antibody domain. In some embodiments, the scFv comprises the V.sub.H antibody domain amino-terminal to the V.sub.L antibody domain. In some embodiments, the scFv comprises the V.sub.L antibody domain amino-terminal to the V.sub.H antibody domain. In some embodiments, there is a peptide linker between the V.sub.L antibody domain and the V.sub.H antibody domain. In some embodiments, all of the V.sub.L antibody domain and V.sub.H antibody domain CDRs are derived from the same antibody moiety. In some embodiments, the V.sub.L antibody domain and the V.sub.H antibody domain comprise antibody CDRs derived from more than one antibody moiety. In some embodiments, the scFv is human, humanized, chimeric, semi-synthetic, or fully synthetic.

[0214] In some embodiments, the TCRM comprises a) a first polypeptide chain comprising a first T cell receptor domain (TCRD) comprising a first transmembrane domain and b) a second polypeptide chain comprising a second TCRD comprising a second transmembrane domain. In some embodiments, the first transmembrane domain is the transmembrane domain of a first TCR subunit and/or the second transmembrane domain is the transmembrane domain of a second TCR subunit. In some embodiments, the first TCR subunit is a TCR chain (e.g., GenBank Accession No: CCI73895), and the second TCR subunit is a TCR chain (e.g., GenBank Accession No: CCI73893). In some embodiments, the first TCR subunit is a TCR chain, and the second TCR subunit is a TCR chain. In some embodiments, the first TCR subunit is a TCR chain (e.g., GenBank Accession No: AGE91788), and the second TCR subunit is a TCR chain (e.g., GenBank Accession No: AAQ57272). In some embodiments, the first TCR subunit is a TCR chain, and the second TCR subunit is a TCR chain. In some embodiments, the first TCRD further comprises a first connecting peptide amino-terminal to the transmembrane domain and/or the second TCRD further comprises a second connecting peptide amino-terminal to the transmembrane domain. In some embodiments, the first connecting peptide comprises all or a portion of the connecting peptide of the first TCR subunit and/or the second connecting peptide comprises all or a portion of the connecting peptide of the second TCR subunit. In some embodiments, the first TCRD further comprises a first TCR intracellular domain carboxy-terminal to the first transmembrane domain and/or the second TCRD further comprises a second TCR intracellular domain carboxy-terminal to the second transmembrane domain. In some embodiments, the first TCR intracellular domain comprises all or a portion of the intracellular domain of the first TCR subunit and/or the second TCR intracellular domain comprises all or a portion of the intracellular domain of the second TCR subunit. In some embodiments, the first TCRD is a fragment of the first TCR subunit and/or the second TCRD is a fragment of the second TCR chain. In some embodiments, the first and second polypeptide chains are linked, such as by a covalent linkage (e.g., peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the first and second TCRDs are linked by a disulfide bond. In some embodiments, the first and second TCRDs are linked by a disulfide bond between a residue in the first connecting peptide and a residue in the second connecting peptide. In some embodiments, the TCRM is capable of recruiting at least one TCR-associated signaling module selected from the group consisting of CD3, CD3, and CD3. In some embodiments, the TCRM is capable of recruiting each of CD3, CD3, and CD3 to form an octameric anti-CD22 caTCR-CD3 complex (i.e., promotes anti-CD22 caTCR-CD3 complex formation).

[0215] In some embodiments, the anti-CD22 caTCR is a molecule comprising a fusion of the antibody moiety (which specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof)) to the TCRM. In some embodiments, the anti-CD22 caTCR comprises a fusion of the first polypeptide chain of the Fab-like or Fv-like antigen-binding module amino-terminal to the first polypeptide chain of the TCRM, thereby forming a first polypeptide chain of the anti-CD22 caTCR, and a fusion of the second polypeptide chain of the Fab-like or Fv-like antigen-binding module amino-terminal to the second polypeptide chain of the TCRM, thereby forming a second polypeptide chain of the anti-CD22 caTCR. In some embodiments, the anti-CD22 caTCR comprises a fusion of the scFv amino-terminal to the first or second polypeptide chain of the TCRM. In some embodiments, the anti-CD22 caTCR further comprises a peptide linker between the first polypeptide chain of the Fab-like or Fv-like antigen-binding module and the first polypeptide chain of the TCRM and/or a peptide linker between the second polypeptide chain of the Fab-like or Fv-like antigen-binding module and the second polypeptide chain of the TCRM. In some embodiments, the anti-CD22 caTCR further comprises a peptide linker between the scFv and the first or second polypeptide chain of the TCRM. In some embodiments, the peptide linker is between about 5 to about 70 (such as about any of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or 70, including any ranges between these values) amino acids in length. In some embodiments, the first polypeptide chain of the anti-CD22 caTCR further comprises an amino-terminal first signal peptide and/or the second polypeptide chain of the anti-CD22 caTCR further comprises an amino-terminal second signal peptide. In some embodiments, the first polypeptide chain of the anti-CD22 caTCR further comprises a first accessory intracellular domain carboxy-terminal to the first transmembrane domain and/or the second polypeptide chain of the anti-CD22 caTCR further comprises a second accessory intracellular domain carboxy-terminal to the second transmembrane domain. In some embodiments, the first and/or second accessory intracellular domains comprise a TCR costimulatory domain. In some embodiments, the first and second polypeptide chains of the anti-CD22 caTCR are linked, such as by a covalent linkage (e.g., peptide or other chemical linkage) or non-covalent linkage. In some embodiments, the anti-CD22 caTCR is a heterodimer.

[0216] Thus, for example, in some embodiments, there is provided an anti-CD22 caTCR comprising a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv.

[0217] In some embodiments, there is provided an anti-CD22 caTCR comprising a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof) comprising i) a light chain variable region comprising LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, and ii) a heavy chain variable region comprising HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively, and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, there is provided an anti-CD22 caTCR comprising a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof) comprising i) a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 206-208, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 212, and ii) a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209-211, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 213, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker; and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv.

[0218] In some embodiments, there is provided an anti-CD22 caTCR comprising a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof) comprising i) a light chain variable region comprising LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, and ii) a heavy chain variable region comprising HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively, and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, there is provided an anti-CD22 caTCR comprising a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof) comprising i) a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 214-216, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 218, and ii) a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209, 210, and 217, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 219, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker; and b) a T cell receptor module (TCRM) capable of recruiting at least one TCR-associated signaling module. In some embodiments, the antibody moiety is a Fab-like antigen-binding module. In some embodiments, the antibody moiety is an Fv-like antigen-binding module. In some embodiments, the antibody moiety is an scFv.

[0219] In some embodiments, the anti-CD22 construct is a chimeric signaling receptor (CSR). For example, the CSR may comprise: (a) an anti-CD22 antibody moiety described herein (e.g., an anti-CD22 antibody moiety comprising a light chain variable region having the sequence of SEQ ID NO: 218 and a heavy chain variable region having the sequence of SEQ ID NO: 219; or an anti-CD22 antibody moiety comprising a light chain variable region having the sequence of SEQ ID NO: 212 and a heavy chain variable region having the sequence of SEQ ID NO: 213); (b) a transmembrane module; and (c) a co-stimulatory immune cell signaling module that is capable of providing a co-stimulatory signal to the immune cell, in which the CSR lacks a functional primary immune cell signaling domain. In some embodiments, the CSR is used in combination with a caTCR or CAR (e.g., a caTCR or CAR that specifically targets CD22).

[0220] Chimeric Co-Stimulatory Receptor (CSR) Constructs

[0221] The ligand-specific chimeric co-stimulatory receptor (CSR) described herein specifically binds to a target ligand (such as a cell surface antigen or a peptide/MHC complex) and is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. The CSR comprises a ligand-binding module that provides the ligand-binding specificity, a transmembrane module, and a co-stimulatory immune cell signaling module that allows for stimulating the immune cell. The CSR lacks a functional primary immune cell signaling sequence. In some embodiments, the CSR lacks any primary immune cell signaling sequence. In some embodiments, the CSR comprises a single polypeptide chain comprising the ligand-binding module, transmembrane module, and co-stimulatory signaling module. In some embodiments, the CSR comprises a first polypeptide chain and a second polypeptide chain, wherein the first and second polypeptide chains together form the ligand-binding module, transmembrane module, and co-stimulatory signaling module. In some embodiments, the first and second polypeptide chains are separate polypeptide chains, and the CSR is a multimer, such as a dimer. In some embodiments, the first and second polypeptide chains are covalently linked, such as by a peptide linkage, or by another chemical linkage, such as a disulfide linkage. In some embodiments, the first polypeptide chain and the second polypeptide chain are linked by at least one disulfide bond. In some embodiments, the expression of the CSR in the caTCR plus CSR immune cell is inducible. In some embodiments, the expression of the CSR in the caTCR plus CSR immune cell is inducible upon signaling through the caTCR. Further description of CSR may be found in U.S. Application No. 62/490,578, filed Apr. 26, 2017, which is incorporated by reference herein in its entirety.

[0222] Examples of co-stimulatory immune cell signaling domains for use in the CSRs of the invention include the cytoplasmic sequences of co-receptors of the T cell receptor (TCR), which can act in concert with a caTCR to initiate signal transduction following caTCR engagement, as well as any derivative or variant of these sequences and any synthetic sequence that has the same functional capability.

[0223] It is known that signals generated through the TCR alone are insufficient for full activation of the T cell and that a secondary or co-stimulatory signal is also required. Thus, T cell activation can be said to be mediated by two distinct classes of intracellular signaling sequence: those that initiate antigen-dependent primary activation through the TCR (referred to herein as primary T cell signaling sequences) and those that act in an antigen-independent manner to provide a secondary or co-stimulatory signal (referred to herein as co-stimulatory T cell signaling sequences).

[0224] Primary immune cell signaling sequences that act in a stimulatory manner may contain signaling motifs which are known as immunoreceptor tyrosine-based activation motifs or ITAMs. Examples of ITAM-containing primary immune cell signaling sequences include those derived from TCR, FcR, FcR, CD3, CD3, CD3, CD5, CD22, CD79a, CD79b, and CD66d. A functional primary immune cell signaling sequence is a sequence that is capable of transducing an immune cell activation signal when operably coupled to an appropriate receptor. Non-functional primary immune cell signaling sequences, which may comprises fragments or variants of primary immune cell signaling sequences, are unable to transduce an immune cell activation signal. The CSRs described herein lack a functional primary immune cell signaling sequence, such as a functional signaling sequence comprising an ITAM. In some embodiments, the CSRs lack any primary immune cell signaling sequence.

[0225] The co-stimulatory immune cell signaling sequence can be a portion of the intracellular domain of a co-stimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like.

[0226] In some embodiments, the target ligand is a cell surface antigen. In some embodiments, the target ligand is a peptide/MHC complex. In some embodiments, the target ligand is the same as the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is different than the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is a molecule presented on the surface of a cell presenting the target antigen. For example, in some embodiments, the target antigen of the caTCR is a cancer-associated antigen presented on a cancer cell, and the target ligand is a ubiquitous molecule expressed on the surface of the cancer cell, such as an integrin. In some embodiments, the target ligand is a disease-associated ligand. In some embodiments, the target ligand is a cancer-associated ligand. In some embodiments, the cancer-associated ligand is, for example, CD19, CD20, CD47, IL4, GPC-3, ROR1, ROR2, BCMA, GPRC5D, or FCRL5. In some embodiments, the cancer-associated ligand is a peptide/MHC complex comprising a peptide derived from a protein including WT-1, AFP, HPV16-E7, NY-ESO-1, PRAME, EBV-LMP2A, and PSA. In some embodiments, the target ligand is a virus-associated ligand. In some embodiments, the target ligand is an immune checkpoint molecule. In some embodiments, the immune checkpoint molecule includes PD-L1, PD-L2, CD80, CD86, ICOSL, B7-H3, B7-H4, HVEM, 4-1BBL, OX40L, CD70, CD40, and GAL9. In some embodiments, the target ligand is an apoptotic molecule. In some embodiments, the apoptotic molecule includes FasL, FasR, TNFR1, and TNFR2.

[0227] In some embodiments, the ligand-binding module is an antibody moiety. In some embodiments, the antibody moiety is a Fab, a Fab, a (Fab)2, an Fv, or a single chain Fv (scFv). In some embodiments, the antibody moiety specifically binds a cell surface antigen, e.g., CD22. In some embodiments, the antibody moiety comprises the CDRs or variables domains (V.sub.H and/or V.sub.L domains) of an antibody moiety specific for CD22 (e.g., V.sub.H domain comprising, consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 219 and/or V.sub.L domain comprising, consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 218, or CDRs contained therein). In some embodiments, the antibody moiety comprises the CDRs or variables domains (V.sub.H and/or V.sub.L domains) of an antibody moiety specific for CD22 (e.g., V.sub.H domain comprising, consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 213 and/or V.sub.L domain comprising, consisting essentially of, or consisting of the amino acid sequence of SEQ ID NO: 212, or CDRs contained therein).

[0228] In some embodiments, the transmembrane module comprises one or more transmembrane domains derived from, for example, CD28, CD3, CD3, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154.

[0229] In some embodiments, the co-stimulatory signaling module comprises, consists essentially of, or consists of all or a portion of the intracellular domain of an immune cell co-stimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like.

[0230] In some embodiments, the CSR further comprises a spacer module between any of the ligand-binding module, the transmembrane module, and the co-stimulatory signaling module. In some embodiments, the spacer module comprises one or more peptide linkers connecting two CSR modules. In some embodiments, the spacer module comprises one or more peptide linkers between about 5 to about 70 (such as about any of 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, or 70, including any ranges between these values) amino acids in length.

[0231] In some embodiments, the ligand-binding module (such as an antibody moiety) specifically binds to a target antigen with a) an affinity that is at least about 10 (including for example at least about any of 10, 20, 30, 40, 50, 75, 100, 200, 300, 400, 500, 750, 1000 or more) times its binding affinity for other molecules; orb) a Kd no more than about 1/10 (such as no more than about any of 1/10, 1/20, 1/30, 1/40, 1/50, 1/75, 1/100, 1/200, 1/300, 1/400, 1/500, 1/750, 1/1000 or less) times its Kd for binding to other molecules. Binding affinity can be determined by methods known in the art, such as ELISA, fluorescence activated cell sorting (FACS) analysis, or radioimmunoprecipitation assay (RIA). Kd can be determined by methods known in the art, such as surface plasmon resonance (SPR) assay utilizing, for example, Biacore instruments, or kinetic exclusion assay (KinExA) utilizing, for example, Sapidyne instruments.

[0232] In some embodiments, the CSR described herein specifically binds to a target ligand (e.g., CD22), comprising a) a target ligand-binding domain (LBD); b) a transmembrane domain; and c) and a co-stimulatory signaling domain, wherein the CSR is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. In some embodiments, the target ligand is a cell surface antigen. In some embodiments, the target ligand is a peptide/MHC complex. In some embodiments, the target ligand is the same as the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is different from the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is a disease-associated ligand. In some embodiments, the target ligand is a cancer-associated ligand. In some embodiments, the cancer-associated ligand is, for example, CD19, CD20, CD47, IL4, GPC-3, ROR1, ROR2, BCMA, GPRCSD, or FCRLS. In some embodiments, the cancer-associated ligand is a peptide/MHC complex comprising a peptide derived from a protein including WT-1, AFP, HPV16-E7, NY-ESO-1, PRAME, EBV-LMP2A, and PSA. In some embodiments, the target ligand is a virus-associated ligand. In some embodiments, the target ligand is an immune checkpoint molecule. In some embodiments, the immune checkpoint molecule includes PD-L1, PD-L2, CD80, CD86, ICOSL, B7-H3, B7-H4, HVEM, 4-1BBL, OX4OL, CD70, CD40, and GAL9. In some embodiments, the target ligand is an apoptotic molecule. In some embodiments, the apoptotic molecule includes FasL, FasR, TNFR1, and TNFR2. In some embodiments, the ligand-binding domain is an antibody moiety. In some embodiments, the antibody moiety is a Fab, a Fab, a (Fab)2, an Fv, or a single chain Fv (scFv). In some embodiments, the ligand-binding domain is (or is derived from) all or a portion of the extracellular domain of a receptor for the target ligand. In some embodiments, the receptor includes, for example, FasR, TNFR1, TNFR2, PD-1, CD28, CTLA-4, ICOS, BTLA, KIR, LAG-3, 4-1BB, OX40, CD27, and TIM-3. In some embodiments, the transmembrane domain comprises a transmembrane domain derived from a transmembrane protein including, for example, CD28, CD3, CD3, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154. In some embodiments, the CSR comprises a fragment of a transmembrane protein (fTMP), wherein the fTMP comprises the CSR transmembrane domain. In some embodiments, the co-stimulatory signaling domain comprises, consists essentially of, or consists of all or a portion of the intracellular domain of an immune cell co-stimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like. In some embodiments, the CSR comprises a fragment of an immune cell co-stimulatory molecule (fCSM), wherein the fCSM comprises the CSR transmembrane domain and CSR co-stimulatory signaling domain. In some embodiments, the CSR further comprises a spacer domain between any of the ligand-binding domain, the transmembrane domain, and the co-stimulatory signaling domain. In some embodiments, the spacer domain comprises a peptide linker connecting two CSR domains.

[0233] In some embodiments, the CSR described herein specifically binds to a target ligand (e.g., CD22), comprising a) a target ligand-binding domain; b) a transmembrane domain; and c) and a co-stimulatory signaling domain, wherein the target ligand is a cell surface antigen, and wherein the CSR is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. In some embodiments, the target ligand is the same as the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is different from the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is a disease-associated ligand. In some embodiments, the target ligand is a cancer-associated ligand. In some embodiments, the cancer-associated ligand is, for example, CD19, CD20, CD47, IL4, GPC-3, ROR1, ROR2, BCMA, GPRC5D, or FCRL5. In some embodiments, the target ligand is a virus-associated ligand. In some embodiments, the target ligand is an immune checkpoint molecule. In some embodiments, the immune checkpoint molecule includes PD-L1, PD-L2, CD80, CD86, ICOSL, B7-H3, B7-H4, HVEM, 4-1BBL, OX4OL, CD70, CD40, and GAL9. In some embodiments, the target ligand is an apoptotic molecule. In some embodiments, the apoptotic molecule includes FasL, FasR, TNFR1, and TNFR2. In some embodiments, the ligand-binding domain is an antibody moiety. In some embodiments, the antibody moiety is a Fab, a Fab, a (Fab)2, an Fv, or a single chain Fv (scFv). In some embodiments, the ligand-binding domain is (or is derived from) all or a portion of the extracellular domain of a receptor for the target ligand. In some embodiments, the receptor includes, for example, FasR, TNFR1, TNFR2, PD-1, CD28, CTLA-4, ICOS, BTLA, KIR, LAG-3, 4-1BB, OX40, CD27, and TIM-3. In some embodiments, the transmembrane domain comprises a transmembrane domain derived from a transmembrane protein including, for example, CD28, CD3, CD3, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154. In some embodiments, the CSR comprises a fragment of a transmembrane protein (fTMP), wherein the fTMP comprises the CSR transmembrane domain. In some embodiments, the co-stimulatory signaling domain comprises, consists essentially of, or consists of all or a portion of the intracellular domain of an immune cell co-stimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like. In some embodiments, the CSR comprises a fragment of an immune cell co-stimulatory molecule (fCSM), wherein the fCSM comprises the CSR transmembrane domain and CSR co-stimulatory signaling domain. In some embodiments, the CSR further comprises a spacer domain between any of the ligand-binding domain, the transmembrane domain, and the co-stimulatory signaling domain. In some embodiments, the spacer domain comprises a peptide linker connecting two CSR domains.

[0234] In some embodiments, the CSR described herein specifically binds to a target ligand (e.g., CD22), comprising a) a target ligand-binding domain; b) a transmembrane domain; and c) and a co-stimulatory signaling domain, wherein the target ligand is a peptide/MHC complex, and wherein the CSR is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. In some embodiments, the target ligand is the same as the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is different from the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is a disease-associated ligand. In some embodiments, the target ligand is a cancer-associated ligand. In some embodiments, the cancer-associated ligand is a peptide/MHC complex comprising a peptide derived from a protein including WT-1, AFP, HPV16-E7, NY-ESO-1, PRAME, EBV-LMP2A, and PSA. In some embodiments, the target ligand is a virus-associated ligand. In some embodiments, the ligand-binding domain is an antibody moiety. In some embodiments, the antibody moiety is a Fab, a Fab, a (Fab)2, an Fv, or a single chain Fv (scFv). In some embodiments, the transmembrane domain comprises a transmembrane domain derived from a transmembrane protein including, for example, CD28, CD3, CD3, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154. In some embodiments, the CSR comprises a fragment of a transmembrane protein (fTMP), wherein the fTMP comprises the CSR transmembrane domain. In some embodiments, the co-stimulatory signaling domain comprises, consists essentially of, or consists of all or a portion of the intracellular domain of an immune cell co-stimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like. In some embodiments, the CSR comprises a fragment of an immune cell co-stimulatory molecule (fCSM), wherein the fCSM comprises the CSR transmembrane domain and CSR co-stimulatory signaling domain. In some embodiments, the CSR further comprises a spacer domain between any of the ligand-binding domain, the transmembrane domain, and the co-stimulatory signaling domain. In some embodiments, the spacer domain comprises a peptide linker connecting two CSR domains.

[0235] In some embodiments, the CSR described herein specifically binds to a target ligand (e.g., CD22), comprising a) a target ligand-binding domain; b) a transmembrane domain; and c) and a co-stimulatory signaling domain, wherein the ligand-binding domain is an antibody moiety, and wherein the CSR is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. In some embodiments, the target ligand is a cell surface antigen. In some embodiments, the target ligand is a peptide/MHC complex. In some embodiments, the target ligand is the same as the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is different from the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is a disease-associated ligand. In some embodiments, the target ligand is a cancer-associated ligand. In some embodiments, the cancer-associated ligand is, for example, CD19, CD20, CD47, IL4, GPC-3, ROR1, ROR2, BCMA, GPRC5D, or FCRL5. In some embodiments, the cancer-associated ligand is a peptide/MHC complex comprising a peptide derived from a protein including WT-1, AFP, HPV16-E7, NY-ESO-1, PRAIVIE, EBV-LMP2A, and PSA. In some embodiments, the target ligand is a virus-associated ligand. In some embodiments, the target ligand is an immune checkpoint molecule. In some embodiments, the immune checkpoint molecule includes PD-L1, PD-L2, CD80, CD86, ICOSL, B7-H3, B7-H4, HVEM, 4-1BBL, OX4OL, CD70, CD40, and GAL9. In some embodiments, the target ligand is an apoptotic molecule. In some embodiments, the apoptotic molecule includes FasL, FasR, TNFR1, and TNFR2. In some embodiments, the antibody moiety is a Fab, a Fab, a (Fab)2, an Fv, or a single chain Fv (scFv). In some embodiments, the transmembrane domain comprises a transmembrane domain derived from a transmembrane protein including, for example, CD28, CD3, CD3, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154. In some embodiments, the CSR comprises a fragment of a transmembrane protein (fTMP), wherein the fTMP comprises the CSR transmembrane domain. In some embodiments, the co-stimulatory signaling domain comprises, consists essentially of, or consists of all or a portion of the intracellular domain of an immune cell co-stimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like. In some embodiments, the CSR comprises a fragment of an immune cell co-stimulatory molecule (fCSM), wherein the fCSM comprises the CSR transmembrane domain and CSR co-stimulatory signaling domain. In some embodiments, the CSR further comprises a spacer domain between any of the ligand-binding domain, the transmembrane domain, and the co-stimulatory signaling domain. In some embodiments, the spacer domain comprises a peptide linker connecting two CSR domains.

[0236] In some embodiments, the CSR described herein specifically binds to a target ligand (e.g., CD22), comprising a) a target ligand-binding domain; b) a transmembrane domain; and c) and a co-stimulatory signaling domain, wherein the ligand-binding domain is (or is derived from) all or a portion of the extracellular domain of a receptor for the target ligand, and wherein the CSR is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. In some embodiments, the target ligand is a cell surface antigen. In some embodiments, the target ligand is the same as the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is different from the target antigen of a caTCR expressed in the same immune cell. In some embodiments, the target ligand is a disease-associated ligand. In some embodiments, the target ligand is a cancer-associated ligand. In some embodiments, the cancer-associated ligand is, for example, CD19, CD20, CD47, IL4, GPC-3, ROR1, ROR2, BCMA, GPRC5D, or FCRL5. In some embodiments, the target ligand is an immune checkpoint molecule. In some embodiments, the immune checkpoint molecule includes PD-L1, PD-L2, CD80, CD86, ICOSL, B7-H3, B7-H4, HVEM, 4-1BBL, OX4OL, CD70, CD40, and GAL9. In some embodiments, the target ligand is an apoptotic molecule. In some embodiments, the apoptotic molecule includes FasL, FasR, TNFR1, and TNFR2. In some embodiments, the target ligand receptor includes, for example, FasR, TNFR1, TNFR2, PD-1, CD28, CTLA-4, ICOS, BTLA, KIR, LAG-3, 4-1BB, OX40, CD27, and TIM-3. In some embodiments, the transmembrane domain comprises a transmembrane domain derived from, for example, CD28, CD3, CD3, CD45, CD4, CD5, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, or CD154. In some embodiments, the co-stimulatory signaling domain comprises, consists essentially of, or consists of all or a portion of the intracellular domain of an immune cell co-stimulatory molecule including, for example, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like. In some embodiments, the CSR further comprises a spacer domain between any of the ligand-binding domain, the transmembrane domain, and the co-stimulatory signaling domain. In some embodiments, the spacer domain comprises a peptide linker connecting two CSR domains.

[0237] In some embodiments, the CSR described herein specifically binds to CD22, comprising an scFv comprising a V.sub.H domain having the amino acid sequence of SEQ ID NO: 219 and a V.sub.L domain having the amino acid sequence of SEQ ID NO: 218. In some embodiments, the scFv comprises, from amino terminus to carboxy terminus, the V.sub.L domain, a peptide linker comprising the amino acid sequence of SEQ ID NO: 233, and the V.sub.H domain.

[0238] In some embodiments, the CSR described herein specifically binds to CD22, comprising an scFv comprising a V.sub.H domain having the amino acid sequence of SEQ ID NO: 213 and a V.sub.L domain having the amino acid sequence of SEQ ID NO: 212. In some embodiments, the scFv comprises, from amino terminus to carboxy terminus, the V.sub.L domain, a peptide linker comprising the amino acid sequence of SEQ ID NO: 233, and the V.sub.H domain.

[0239] In some embodiments, a CSR can be an anti-CD22 CSR, an anti-CD19 CSR, or an anti-CD20 CSR. The CSR can comprise an intracellular fragment (i.e., a co-stimulatory immune cell signaling sequence), which can be a portion of the intracellular domain of a co-stimulatory molecule. Examples of co-stimulatory molecules include, but are not limited to, CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, a ligand that specifically binds with CD83, and the like. In some embodiments, the CSR can comprise a transmembrane fragment, which can be a fragment of the transmembrane portion of the co-stimulatory molecule. In some embodiments, the intracellular fragment and the transmembrane fragment in the CSR can be from the same co-stimulatory molecule (e.g., CD28 or CD30). In some embodiments, the intracellular fragment and the transmembrane fragment in the CSR can be from the different co-stimulatory molecules. In some embodiments, the intracellular fragment can be taken from a first co-stimulatory molecule and fused to the transmembrane fragment from a second co-stimulatory molecule. For example, the transmembrane fragment can be from the transmembrane portion of CD8 and the intracellular fragment can be from the intracellular domain of CD30. In some embodiments, when the intracellular fragment and the transmembrane fragment in the CSR are from the same co-stimulatory molecule, the intracellular fragment and the transmembrane fragment can be taken from a single co-stimulatory molecule (e.g., a single CD28 or a single CD30). In other embodiments, when the intracellular fragment and the transmembrane fragment in the CSR are from the same co-stimulatory molecule, the intracellular fragment can be taken from a first co-stimulatory molecule (e.g., a first CD28) and fused to the transmembrane fragment taken from a second co-stimulatory molecule (e.g., a second CD28).

[0240] In some embodiments, an anti-CD19 CSR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96. In some embodiments, an anti-CD22 CSR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112. In some embodiments, an anti-CD20 CSR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128. In some embodiments, a CSR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0241] In some embodiments, the expression of the CSR in the caTCR plus CSR immune cell is inducible. In some embodiments, the caTCR plus CSR immune cell comprises a nucleic acid sequence encoding the CSR operably linked to an inducible promoter, including any of the inducible promoters described herein. In some embodiments, the expression of the CSR in the caTCR plus CSR immune cell is inducible upon signaling through the caTCR. In some such embodiments, the caTCR plus CSR immune cell comprises a nucleic acid sequence encoding the CSR operably linked to a promoter or regulatory element responsive to signaling through the caTCR. In some embodiments, the nucleic acid sequence encoding the CSR is operably linked to a nuclear-factor of the activated T-cell (NFAT)-derived promoter. In some embodiments, the NFAT-derived promoter is an NFAT-derived minimal promoter (see for example Durand, D. et. al., Molec. Cell. Biol. 8, 1715-1724 (1988); Clipstone, N A, Crabtree, G R. Nature. 1992 357(6380): 695-7; Chmielewski, M., et al. Cancer research 71.17 (2011): 5697-5706; and Zhang, L., et al. Molecular therapy 19.4 (2011): 751-759).

[0242] Also provided herein are effector cells (such as lymphocytes, e.g., T cells) expressing an anti-CD22 construct, such as an anti-CD22 CAR, anti-CD22 caTCR, or anti-CD22 CSR.

[0243] Also provided is a method of producing an effector cell expressing an anti-CD22 CAR, anti-CD22 caTCR, or anti-CD22 CSR, the method comprising introducing a vector comprising a nucleic acid encoding the anti-CD22 CAR, anti-CD22 caTCR, or anti-CD22 CSR into the effector cell. In some embodiments, introducing the vector into the effector cell comprises transducing the effector cell with the vector. In some embodiments, introducing the vector into the effector cell comprises transfecting the effector cell with the vector. Transduction or transfection of the vector into the effector cell can be carried about using any method known in the art.

[0244] Monospecific Constructs

[0245] The invention features monospecific constructs that specifically bind to a target ligand (such as a cell surface antigen or a peptide/MHC complex) and are capable of stimulating an immune cell on the surface of which they are functionally expressed upon target ligand binding. In some embodiments, the monospecific construct specifically targets CD22. In some embodiments, a monospecific construct can be a caTCR, a CSR, a CAR, a full-length antibody, a Fab, a Fab, a F(ab)2, an Fv, a single chain Fv (scFv) antibody, a tandem scFv, a diabody (Db), a single chain diabody (scDb), a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, a knob-into-hole (KiH) antibody, a dock and lock (DNL) antibody, a chemically cross-linked antibody, a heteromultimeric antibody, or a heteroconjugate antibody.

[0246] In some embodiments, a monospecific construct is an anti-CD22 construct comprising an anti-CD22 antibody moiety. In certain embodiments, the anti-CD22 antibody moiety comprises: (a) the light chain variable region (VL) comprising a sequence having at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 212; and (b) a heavy chain variable region (VH) comprising a sequence having at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 213. In certain embodiments, the anti-CD22 antibody moiety comprises: (a) the light chain variable region (V.sub.L ) comprising a sequence having at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 218; and (b) a heavy chain variable region (VH) comprising a sequence having at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 219. In other embodiments, the anti-CD22 antibody moiety comprises an anti-CD22 scFv comprising a sequence having at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 186.

[0247] In some embodiments, a monospecific anti-CD22 construct is a caTCR, a CSR, a CAR, or an antibody, each of which can be monovalent or multivalent (e.g., bivalent, trivalent, quad-valent). For example, construct 4 (SEQ ID NO: 4) comprises an anti-CD22 scFv and an anti-CD22 Fab and is also referred to as a bivalent-monospecific construct. In some embodiments, a monospecific anti-CD22 construct is a multivalent caTCR, which can contain multiple Fabs or combiantions of Fabs and scFvs.

[0248] In some embodiments, a monospecific construct can be a caTCR, e.g., an anti-CD22 caTCR, an anti-CD19 caTCR, or an anti-CD20 caTCR. In some embodiments, a monospecific caTCR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 2, 4, 22, 28, and 76-80.

[0249] In some embodiments, a monospecific construct can be a CSR, e.g., an anti-CD22 CSR, an anti-CD19 CSR, or an anti-CD20 CSR. In some embodiments, an anti-CD19 CSR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96. In some embodiments, an anti-CD22 CSR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112. In some embodiments, an anti-CD20 CSR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0250] Multispecific Constructs

[0251] In some embodiments, the anti-CD22 constructs can be multispecific (e.g., bispecific). Multispecific anti-CD22 constructs have antibody moieties against more than one target. In some embodiments, a multispecific anti-CD22 construct has one antibodity moiety against CD22 and one or more additional antibody moieties against one or more non-CD22 antigens (e.g., CD19, CD20, a non-CD22 antigen expressed in B-cell malignancy). In some embodiments, a multispecific anti-CD22 construct can be bispecific, trispecific, quad-specific, or quint-specific. In some embodiments, a multispecific anti-CD22 construct can be a caTCR, a CSR, a CAR, a full-length antibody, a Fab, a Fab, a F(ab)2, an Fv, a single chain Fv (scFv) antibody, a tandem scFv, a diabody (Db), a single chain diabody (scDb), a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, a knob-into-hole (KiH) antibody, a dock and lock (DNL) antibody, a chemically cross-linked antibody, a heteromultimeric antibody, or a heteroconjugate antibody.

[0252] Multispecific ConstructsBispecific Constructs

[0253] The invention features bispecific constructs that specifically bind to two target ligands (such as cell surface antigens or peptide/MHC complexes) and are capable of stimulating an immune cell on the surface of which they are functionally expressed upon target ligand binding. In some embodiments, the bispecific construct specifically targets CD22 and CD19. In other embodiments, the bispecific construct specifically targets CD22 and CD20.

[0254] In some embodiments, a bispecific construct can be a caTCR, e.g., an anti-CD22-anti-CD19 caTCR. In some embodiments, a bispecific caTCR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 15-21.

[0255] In some embodiments, a bispecific construct can be a CSR. In some embodiments, a bispecific CSR comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0256] Multispecific ConstructsTrispecific Constructs

[0257] The invention features trispecific constructs that specifically bind to three target ligands (such as cell surface antigens or peptide/MHC complexes) and are capable of stimulating an immune cell on the surface of which they are functionally expressed upon target ligand binding. In some embodiments, the trispecific construct specifically targets CD22, CD19, and CD20.

[0258] In some embodiments, a trispecific construct comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 24-27. In some embodiments, the FLGA tag (SEQ ID NO: 195) is removed from the sequence of any one of SEQ ID NOS: 24-27.

[0259] Construct Combinations

[0260] The invention features construct combinations comprising at least two different constructs of the invention. In some embodiments, the at least two different constructs are of the same kind of constructs, e.g., two different antibodies (e.g., two different full-length IgG or two different bispecific antibodies), two different CARs, two different caTCRs, or two different CSRs. In some embodiments, the at least two different constructs are of different kinds of constructs, e.g., an antibody plus a CAR, an antibody plus a caTCR, a CAR plus a CSR, a caTCR plus a CSR, etc. In some embodiments, the construct combination comprises at least one monospecific construct. In some embodiments, the construct combination comprises at least one multispecific construct. In some embodiments, the construct combination comprises at least one monospecific construct and at least one multispecific construct. In some embodiments, the at least two different constructs in the construct combination have antibody moities against at least one common target. In some embodiments, the at least two different constructs in the construct combination do not have antibody moities against any common targets.

[0261] In some embodiments, the construct combination comprises at least one anti-CD22 construct of the invention. In some embodiments, the at least one anti-CD22 construct is an antibody, a CAR, a caTCR, or a CSR. In some embodiments, the at least one anti-CD22 construct is a monospecific construct. In some embodiments, the at least one anti-CD22 construct is a multispecific construct. In some embodiments, the at least one anti-CD22 construct is an anti-CD22 caTCR, and the construct combination comprises a CSR. In some embodiments, the anti-CD22 caTCR is expressed in combination with the CSR. In some embodiments, the at least one anti-CD22 construct is an anti-CD22 CSR, and the construct combination comprises a caTCR. In some embodiments, the anti-CD22 CSR is expressed in combination with the caTCR.

[0262] Construct Combinations Comprising Monospecific Constructs

[0263] In some embodiments, the construct combination comprises at least one monospecific construct which specifically binds to a target ligand (such as a cell surface antigen or a peptide/WIC complex) and is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. In some embodiments, the monospecific construct specifically targets CD22.

[0264] In some embodiments, the construct combination comprises a monospecific construct, and the construct combination comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 1, 3, and 5-14.

[0265] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 77 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0266] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 77 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0267] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 77 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0268] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 78 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0269] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 78 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0270] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 78 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0271] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 77 and the CSR has a sequence that comprises a transmembrane fragment of CD8, 4-1BB, CD27, CD28, CD30, or OX40. In some embodiments, the transmembrane fragment has a sequence of any one of SEQ ID NOS: 145-150. In some embodimetns, the CSR comprises an anti-CD22 moiety.

[0272] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 77 and the CSR has a sequence that comprises an intracellular fragment of a molecule selected from the group consisting of CD28, 4-1BB (CD137), OX40, CD30, and CD27. In some embodiments, the intracellular fragment comprises a sequence of any one of SEQ ID NOS: 151-155. In some embodimetns, the CSR comprises an anti-CD22 moiety.

[0273] In some embodiments, the construct combination comprises a monospecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 77 and the CSR has a sequence that comprises a sequence of any one of SEQ ID NOS: 156-171. In some embodimetns, the CSR comprises an anti-CD22 moiety.

[0274] In any one of the construct combinations disclosed herein, in some embodimetns, each caTCR and CSR can be encoded by a single nucleic acid, expressed, and translated as a single polypeptide initially, and then get cleaved into separate polypeptides. The caTCR and the CSR can be connected via a cleavable P2A peptide (e.g., SEQ ID NO: 190). In other embodiments, the caTCR and the CSR can be encoded by two different nucleic acids, expressed, and separately translated separate polypeptides.

[0275] Construct Combinations Comprising Bispecific Constructs

[0276] In some embodiments, the construct combination comprises at least one bispecific construct which specifically binds to two target ligands (such as cell surface antigens or peptide/MHC complexes) and is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. In some embodiments, the bispecific construct specifically targets CD22 and CD19. In other embodiments, the bispecific construct specifically targets CD22 and CD20.

[0277] In some embodiments, the construct combination comprises a bispecific construct, and the construct combination comprises a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 23 and 29-75.

[0278] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 15 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0279] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 16 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0280] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 17 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0281] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 18 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0282] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 19 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0283] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 20 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0284] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 21 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0285] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 15 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0286] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 16 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0287] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 17 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0288] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 18 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0289] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 19 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0290] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 20 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0291] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 21 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0292] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 15 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0293] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 16 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0294] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 17 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0295] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 18 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0296] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 19 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0297] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 20 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0298] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 21 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0299] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 15 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0300] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 16 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0301] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 17 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0302] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 18 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0303] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 19 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0304] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 20 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0305] In some embodiments, the construct combination comprises a bispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 21 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 129-144.

[0306] In any one of the construct combinations comprising a bispecific caTCR and a CSR disclosed herein, in some embodimetns, each caTCR and CSR can be encoded by a single nucleic acid, expressed, and translated as a single polypeptide initially, and then get cleaved into separate polypeptides. The caTCR and the CSR can be connected via a cleavable P2A peptide (e.g., SEQ ID NO: 190). In other embodiments, the caTCR and the CSR can be encoded in two different nucleic acids, expressed, and separately translated separate polypeptides.

[0307] Construct Combinations Comprising Trispecific Constructs

[0308] In some embodiments, the construct combination comprises at least one trispecific construct which specifically binds to three target ligands (such as cell surface antigens or peptide/MHC complexes) and is capable of stimulating an immune cell on the surface of which it is functionally expressed upon target ligand binding. In some embodiments, the trispecific construct combination specifically targets CD22, CD19, and CD20.

[0309] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 24 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0310] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 25 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0311] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 26 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0312] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 27 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96.

[0313] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 24 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0314] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 25 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0315] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 26 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0316] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 27 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112.

[0317] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 24 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0318] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 25 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0319] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 26 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0320] In some embodiments, the construct combination comprises a trispecific caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 27 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0321] In any one of the construct combinations comprising a trispecific caTCR and a CSR disclosed herein, in some embodimetns, each caTCR and CSR can be encoded by a single nucleic acid, expressed, and translated as a single polypeptide initially, and then get cleaved into separate polypeptides. The caTCR and the CSR can be connected via a cleavable P2A peptide (e.g., SEQ ID NO: 190). In other embodiments, the caTCR and the CSR can be encoded in two different nucleic acids, expressed, and separately translated separate polypeptides.

[0322] In some embodiments, a caTCR, when associated with CD3t upon binding target antigen, can activate T cells. A CSR generally does not associate with CD3 and does not activate T cells upon binding target antigen. Additional description and examples of caTCRs and CSRs can be found in International Patent Publication Nos. WO2018/200582 and WO2018/200583, each of which is incorproated herein by reference in its entirety.

[0323] Antibody Drug Conjugates

[0324] In some embodiments, there is provided an anti-CD22 immunoconjugate comprising an anti-CD22 antibody moiety and a therapeutic agent (also referred to herein as an antibody-drug conjugate, or ADC). In some embodiments, the therapeutic agent is a toxin that is either cytotoxic, cytostatic, or otherwise prevents or reduces the ability of the target cells to divide. The use of ADCs for the local delivery of cytotoxic or cytostatic agents, i.e., drugs to kill or inhibit tumor cells in the treatment of cancer (Syrigos and Epenetos, Anticancer Research 19: 605-614 (1999); Niculescu-Duvaz and Springer, Adv. Drg. Del. Rev. 26: 151-172 (1997); U.S. Pat. No. 4,975,278) allows targeted delivery of the drug moiety to target cells, and intracellular accumulation therein, where systemic administration of these unconjugated therapeutic agents may result in unacceptable levels of toxicity to normal cells as well as the target cells sought to be eliminated (Baldwin et al., Lancet (Mar. 15, 1986): 603-605 (1986); Thorpe, (1985) Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review, in Monoclonal Antibodies '84: Biological And Clinical Applications, A. Pinchera et al. (eds.), pp. 475- 506). Maximal efficacy with minimal toxicity is sought thereby.

[0325] Therapeutic agents used in anti-CD22 immunoconjugates (e.g., an anti-CD22 ADC) include, for example, daunomycin, doxorubicin, methotrexate, and vindesine (Rowland et al., Cancer Immunol. Immunother. 21: 183-187 (1986)). Toxins used in anti-CD22 immunoconjugates include bacterial toxins such as diphtheria toxin, plant toxins such as ricin, small molecule toxins such as geldanamycin (Mandler et al., J. Nat. Cancer Inst. 92(19): 1573-1581 (2000); Mandler et al., Bioorganic & Med. Chem. Letters 10: 1025- 1028 (2000); Mandler et al., Bioconjugate Chem. 13: 786-791 (2002)), maytansinoids (EP 1391213; Liu et al., Proc. Natl. Acad. Sci. USA 93: 8618-8623 (1996)), and calicheamicin (Lode et al., Cancer Res. 58: 2928 (1998); Hinman et al., Cancer Res. 53: 3336-3342 (1993)). The toxins may exert their cytotoxic and cytostatic effects by mechanisms including tubulin binding, DNA binding, or topoisomerase inhibition. Some cytotoxic drugs tend to be inactive or less active when conjugated to large antibodies or protein receptor ligands.

[0326] Enzymatically active toxins and fragments thereof that can be used include, for example, diphtheria A chain, nonbinding active fragments of diphtheria toxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain, abrin A chain, modeccin A chain, -sarcin, Aleurites fordii proteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII, and PAP-S), momordica charantia inhibitor, curcin, crotin, sapaonaria officinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin, enomycin, and the tricothecenes. See, e.g., WO 93/21232 published Oct. 28, 1993.

[0327] Anti-CD22 immunoconjugates (e.g., an anti-CD22 ADC) of an anti-CD22 antibody moiety and one or more small molecule toxins, such as a calicheamicin, maytansinoids, dolastatins, aurostatins, a trichothecene, and CC1065, and the derivatives of these toxins that have toxin activity, are also contemplated herein.

[0328] In some embodiments, there is provided an anti-CD22 immunoconjugate (e.g., an anti-CD22 ADC) comprising a therapeutic agent that has an intracellular activity. In some embodiments, the anti-CD22 immunoconjugate is internalized and the therapeutic agent is a cytotoxin that blocks the protein synthesis of the cell, therein leading to cell death. In some embodiments, the therapeutic agent is a cytotoxin comprising a polypeptide having ribosome-inactivating activity including, for example, gelonin, bouganin, saporin, ricin, ricin A chain, bryodin, diphtheria toxin, restrictocin, Pseudomonas exotoxin A and variants thereof. In some embodiments, where the therapeutic agent is a cytotoxin comprising a polypeptide having a ribosome-inactivating activity, the anti-CD22 immunoconjugate must be internalized upon binding to the target cell in order for the protein to be cytotoxic to the cells.

[0329] In some embodiments, there is provided an anti-CD22 immunoconjugate (e.g., an anti-CD22 ADC) comprising a therapeutic agent that acts to disrupt DNA. In some embodiments, the therapeutic agent that acts to disrupt DNA is, for example, selected from the group consisting of enediyne (e.g., calicheamicin and esperamicin) and non-enediyne small molecule agents (e.g., bleomycin, methidiumpropyl-EDTA-Fe(II)).

[0330] The present invention further contemplates an anti-CD22 immunoconjugate (e.g., an anti-CD22 ADC) formed between the anti-CD22 antibody moiety and a compound with nucleolytic activity (e.g., a ribonuclease or a DNA endonuclease such as a deoxyribonuclease; DNase).

[0331] In some embodiments, the anti-CD22 immunoconjugate comprises an agent that acts to disrupt tubulin. Such agents may include, for example, rhizoxin/maytansine, paclitaxel, vincristine and vinblastine, colchicine, auristatin dolastatin 10 MMAE, and peloruside A.

[0332] In some embodiments, the anti-CD22 immunoconjugate (e.g., an anti-CD22 ADC) comprises an alkylating agent including, for example, Asaley NSC 167780, AZQ NSC 182986, BCNU NSC 409962, Busulfan NSC 750, carboxyphthalatoplatinum NSC 271674, CBDCA NSC 241240, CCNU NSC 79037, CHIP NSC 256927, chlorambucil NSC 3088, chlorozotocin NSC 178248, cis-platinum NSC 119875, clomesone NSC 338947, cyanomorpholinodoxorubicin NSC 357704, cyclodisone NSC 348948, dianhydrogalactitol NSC 132313, fluorodopan NSC 73754, hepsulfam NSC 329680, hycanthone NSC 142982, melphalan NSC 8806, methyl CCNU NSC 95441 , mitomycin C NSC 26980, mitozolamide NSC 353451 , nitrogen mustard NSC 762, PCNU NSC 95466, piperazine NSC 344007, piperazinedione NSC 135758, pipobroman NSC 25154, porfiromycin NSC 56410, spirohydantoin mustard NSC 172112, teroxirone NSC 296934, tetraplatin NSC 363812, thio-tepa NSC 6396, triethylenemelamine NSC 9706, uracil nitrogen mustard NSC 34462, and Yoshi-864 NSC 102627.

[0333] In some embodiments, the anti-CD22 immunoconjugate (e.g., an anti-CD22 ADC) comprises a highly radioactive atom. A variety of radioactive isotopes are available for the production of radioconjugated antibodies. Examples include 211At, 1311, 1251, 90Y, 186Re, 188Re, 153Sm, 212Bi, 32P, 212Pb and radioactive isotopes of Lu.

[0334] In some embodiments, the anti-CD22 antibody moiety can be conjugated to a receptor (such as streptavidin) for utilization in tumor pre-targeting wherein the antibody-receptor conjugate is administered to the patient, followed by removal of unbound conjugate from the circulation using a clearing agent and then administration of a ligand (e.g., avidin) that is conjugated to a cytotoxic agent (e.g., a radionucleotide).

[0335] In some embodiments, an anti-CD22 immunoconjugate (e.g., an anti-CD22 ADC) may comprise an anti-CD22 antibody moiety conjugated to a prodrug-activating enzyme. In some such embodiments, a prodrug-activating enzyme converts a prodrug to an active drug, such as an anti-viral drug. Such anti-CD22 immunoconjugates are useful, in some embodiments, in antibody-dependent enzyme-mediated prodrug therapy (ADEPT). Enzymes that may be conjugated to an antibody include, but are not limited to, alkaline phosphatases, which are useful for converting phosphate-containing prodrugs into free drugs; arylsulfatases, which are useful for converting sulfate-containing prodrugs into free drugs; proteases, such as serratia protease, thermolysin, subtilisin, carboxypeptidases and cathepsins (such as cathepsins B and L), which are useful for converting peptide-containing prodrugs into free drugs; D-alanylcarboxypeptidases, which are useful for converting prodrugs that contain D-amino acid substituents; carbohydrate-cleaving enzymes such as -galactosidase and neuraminidase, which are useful for converting glycosylated prodrugs into free drugs; -lactamase, which is useful for converting drugs derivatized with -lactams into free drugs; and penicillin amidases, such as penicillin V amidase and penicillin G amidase, which are useful for converting drugs derivatized at their amine nitrogens with phenoxyacetyl or phenylacetyl groups, respectively, into free drugs. In some embodiments, enzymes may be covalently bound to antibody moieties by recombinant DNA techniques well known in the art. See, e.g., Neuberger et al., Nature 312: 604-608 (1984).

[0336] In some embodiments, the therapeutic portion of the anti-CD22 immunoconjugates (e.g., an anti-CD22 ADC) may be a nucleic acid. Nucleic acids that may be used include, but are not limited to, anti-sense RNA, genes or other polynucleotides, including nucleic acid analogs such as thioguanine and thiopurine.

[0337] The present application further provides anti-CD22 immunoconjugates (e.g., an anti-CD22 ADC) comprising an anti-CD22 antibody moiety attached to an effector molecule, wherein the effector molecule is a label, which can generate a detectable signal, indirectly or directly. These anti-CD22 immunoconjugates can be used for research or diagnostic applications, such as for the in vivo detection of cancer. The label is preferably capable of producing, either directly or indirectly, a detectable signal. For example, the label may be radio-opaque or a radioisotope, such as 3H, 14C, 32P, 35S, 123I, 125I, 131I; a fluorescent (fluorophore) or chemiluminescent (chromophore) compound, such as fluorescein isothiocyanate, rhodamine or luciferin; an enzyme, such as alkaline phosphatase,-galactosidase or horseradish peroxidase; an imaging agent; or a metal ion. In some embodiments, the label is a radioactive atom for scintigraphic studies, for example 99Tc or 123I, or a spin label for nuclear magnetic resonance (NMR) imaging (also known as magnetic resonance imaging, MRI), such as zirconium-89, iodine-123, iodine-131, indium-111, fluorine-19, carbon-13, nitrogen-15, oxygen-17, gadolinium, manganese or iron. Zirconium-89 may be complexed to various metal chelating agents and conjugated to antibodies, e.g., for PET imaging (WO 2011/056983).

[0338] In some embodiments, the anti-CD22 immunoconjugate is detectable indirectly. For example, a secondary antibody that is specific for the anti-CD22 immunoconjugate and contains a detectable label can be used to detect the anti-CD22 immunoconjugate.

[0339] III. caTCR Plus CSR Immune Cells

[0340] The present invention provides an immune cell (such as a T cell) presenting on its surface a caTCR and a CSR according to any of the caTCRs and CSRs described herein (such an immune cell is also referred to herein as a caTCR plus CSR immune cell). In some embodiments, the immune cell comprises nucleic acid encoding the caTCR and CSR, wherein the caTCR and CSR are expressed from the nucleic acid and localized to the immune cell surface. In some embodiments, the immune cell is a T cell. In some embodiments, the immune cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell. In some embodiments, the immune cell does not express the TCR subunits from which the TCR-TMs of the caTCR are derived. For example, in some embodiments, the immune cell is an T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and chains, or the T cell is a T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and chains. In some embodiments, the immune cell is modified to block or decrease the expression of one or both of the endogenous TCR subunits of the immune cell. For example, in some embodiments, the immune cell is an T cell modified to block or decrease the expression of the TCR and/or chains or the immune cell is a T cell modified to block or decrease the expression of the TCR and/or chains. Modifications of cells to disrupt gene expression include any such techniques known in the art, including for example RNA interference (e.g., siRNA, shRNA, miRNA), gene editing (e.g., CRISPR- or TALEN-based gene knockout), and the like.

[0341] For example, in some embodiments, there is provided an immune cell (such as a T cell) comprising nucleic acid encoding a caTCR according to any of the caTCRs described herein and a CSR according to any of the CSRs described herein, wherein the caTCR and CSR are expressed from the nucleic acid and localized to the immune cell surface. In some embodiments, the nucleic acid comprises a first caTCR nucleic acid sequence encoding a first caTCR polypeptide chain of the caTCR, a second caTCR nucleic acid sequence encoding a second caTCR polypeptide chain of the caTCR, and a CSR nucleic acid sequence encoding a CSR polypeptide chain of the CSR. In some embodiments, the first and second caTCR nucleic acid sequences and CSR nucleic acid sequence are each contained in different vectors. In some embodiments, some or all of the nucleic acid sequences are contained in the same vector. Vectors may be selected, for example, from the group consisting of mammalian expression vectors and viral vectors (such as those derived from retroviruses, adenoviruses, adeno-associated viruses, herpes viruses, and lentiviruses). In some embodiments, one or more of the vectors is integrated into the host genome of the immune cell. In some embodiments, the first and second caTCR nucleic acid sequences and CSR nucleic acid sequence are each under the control of different promoters. In some embodiments, some or all of the promoters have the same sequence. In some embodiments, some or all of the promoters have different sequences. In some embodiments, some or all of the nucleic acid sequences are under the control of a single promoter. In some embodiments, some or all of the promoters are inducible. In some embodiments, the immune cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell.

[0342] Thus, in some embodiments, there is provided a caTCR plus CSR immune cell (such as a T cell) expressing on its surface a caTCR according to any of the caTCRs described herein and a CSR according to any of the CSRs described herein, wherein the caTCR plus CSR immune cell comprises a) a first caTCR nucleic acid sequence encoding a first caTCR polypeptide chain of the caTCR; b) a second caTCR nucleic acid sequence encoding a second caTCR polypeptide chain of the caTCR; and c) a CSR nucleic acid sequence encoding a CSR polypeptide chain of the CSR, wherein the first and second caTCR polypeptide chains are expressed from the first and second caTCR nucleic acid sequences to form the caTCR, wherein the CSR polypeptide chain is expressed from the CSR nucleic acid to form the CSR, and wherein the caTCR and CSR localize to the surface of the immune cell. In some embodiments, the first caTCR nucleic acid sequence is contained in a first vector (such as a lentiviral vector), the second caTCR nucleic acid sequence is contained in a second vector (such as a lentiviral vector), and the CSR nucleic acid sequence is contained in a third vector (such as a lentiviral vector). In some embodiments, some or all of the first and second caTCR nucleic acid sequences and CSR nucleic acid sequence are contained in the same vector (such as a lentiviral vector). In some embodiments, each of the first and second caTCR nucleic acid sequences and CSR nucleic acid sequence are, individually, operably linked to a promoter. In some embodiments, some or all of the nucleic acid sequences are under the control of a single promoter. In some embodiments, some or all of the promoters have the same sequence. In some embodiments, some or all of the promoters have different sequences. In some embodiments, some or all of the promoters are inducible. In some embodiments, some or all of the vectors are viral vectors (such as lentiviral vectors). In some embodiments, the immune cell does not express the TCR subunits from which the TCR-TMs of the caTCR are derived. For example, in some embodiments, the immune cell is an T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and chains, or the immune cell is a T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and 0 chains. In some embodiments, the immune cell is modified to block or decrease the expression of one or both of its endogenous TCR subunits. For example, in some embodiments, the immune cell is an T cell modified to block or decrease the expression of the TCR and/or chains, or the immune cell is a T cell modified to block or decrease the expression of the TCR and/or chains. In some embodiments, the immune cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell. In some embodiments, some or all of the vectors are viral vectors (such as lentiviral vectors) integrated into the host genome of the immune cell.

[0343] In some embodiments, there is provided a caTCR plus CSR immune cell (such as a T cell) expressing on its surface a caTCR according to any of the caTCRs described herein and a CSR according to any of the CSRs described herein, wherein the caTCR plus CSR immune cell comprises a) a first vector comprising a first promoter operably linked to a first caTCR nucleic acid sequence encoding a first caTCR polypeptide chain of the caTCR; b) a second vector comprising a second promoter operably linked to a second caTCR nucleic acid sequence encoding a second caTCR polypeptide chain of the caTCR; and c) a third vector comprising a third promoter operably linked to a CSR nucleic acid sequence encoding a CSR polypeptide chain of the CSR, wherein the first and second caTCR polypeptide chains are expressed from the first and second caTCR nucleic acid sequences to form the caTCR and the CSR polypeptide chain is expressed from the CSR nucleic acid sequence to form the CSR, and wherein the caTCR and CSR localize to the surface of the immune cell. In some embodiments, some or all of the promoters have the same sequence. In some embodiments, some or all of the promoters have different sequences. In some embodiments, some or all of the promoters are inducible. In some embodiments, the immune cell does not express the TCR subunits from which the TCR-TMs of the caTCR are derived. For example, in some embodiments, the immune cell is an T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and chains, or the immune cell is a T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR a and 0 chains. In some embodiments, the immune cell is modified to block or decrease the expression of one or both of its endogenous TCR subunits. For example, in some embodiments, the immune cell is an T cell modified to block or decrease the expression of the TCR a and/or 0 chains, or the immune cell is a T cell modified to block or decrease the expression of the TCR and/or chains. In some embodiments, the immune cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell. In some embodiments, the first and second vectors are viral vectors (such as lentiviral vectors) integrated into the host genome of the immune cell.

[0344] In some embodiments, there is provided a caTCR plus CSR immune cell (such as a T cell) expressing on its surface a caTCR according to any of the caTCRs described herein and a CSR according to any of the CSRs described herein, wherein the caTCR plus CSR immune cell comprises a) a first vector comprising i) a first promoter operably linked to a first caTCR nucleic acid sequence encoding a first caTCR polypeptide chain of the caTCR and ii) a second promoter operably linked to a second caTCR nucleic acid sequence encoding a second caTCR polypeptide chain of the caTCR; and b) a second vector comprising a third promoter operably linked to a CSR nucleic acid sequence encoding a CSR polypeptide chain of the CSR, wherein the first and second caTCR polypeptide chains are expressed from the first and second caTCR nucleic acid sequences to form the caTCR and the CSR polypeptide chain is expressed from the CSR nucleic acid sequence to form the CSR, and wherein the caTCR localizes to the surface of the immune cell. In some embodiments, some or all of the promoters have the same sequence. In some embodiments, some or all of the promoters have different sequences. In some embodiments, some or all of the promoters are inducible. In some embodiments, the immune cell does not express the TCR subunits from which the TCR-TMs of the caTCR are derived. For example, in some embodiments, the immune cell is an T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and chains, or the immune cell is a T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and chains. In some embodiments, the immune cell is modified to block or decrease the expression of one or both of its endogenous TCR subunits. For example, in some embodiments, the immune cell is an T cell modified to block or decrease the expression of the TCR and/or chains, or the immune cell is a T cell modified to block or decrease the expression of the TCR and/or chains. In some embodiments, the immune cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell. In some embodiments, the first and second vectors are viral vectors (such as lentiviral vectors) integrated into the host genome of the immune cell. It is to be appreciated that embodiments where any of the nucleic acid sequences are swapped are also contemplated, such as where the first or second caTCR nucleic acid sequence is swapped with the CSR nucleic acid sequence.

[0345] In some embodiments, there is provided a caTCR plus CSR immune cell (such as a T cell) expressing on its surface a caTCR according to any of the caTCRs described herein and a CSR according to any of the CSRs described herein, wherein the caTCR plus CSR immune cell comprises a) a first vector comprising i) a first caTCR nucleic acid sequence encoding a first caTCR polypeptide chain of the caTCR and ii) a second caTCR nucleic acid sequence encoding a second caTCR polypeptide chain of the caTCR, wherein the first and second caTCR nucleic acid sequences are under the control of a first promoter; and b) a second vector comprising a second promoter operably linked to a CSR nucleic acid sequence encoding a CSR polypeptide chain of the CSR, wherein the first and second caTCR polypeptide chains are expressed from the first and second caTCR nucleic acid sequences to form the caTCR and the CSR polypeptide chain is expressed from the CSR nucleic acid sequence to form the CSR, and wherein the caTCR and CSR localize to the surface of the immune cell. In some embodiments, the first promoter is operably linked to the 5 end of the first caTCR nucleic acid sequence, and there is nucleic acid linker selected from the group consisting of an internal ribosomal entry site (IRES) and a nucleic acid encoding a self-cleaving 2A peptide (such as P2A, T2A, E2A, or F2A) linking the 3 end of first caTCR nucleic acid sequence to the 5 end of the second caTCR nucleic acid sequence, wherein the first caTCR nucleic acid sequence and the second caTCR nucleic acid sequence are transcribed as a single RNA under the control of the promoter. In some embodiments, the first promoter is operably linked to the 5 end of the second caTCR nucleic acid sequence, and there is nucleic acid linker selected from the group consisting of an internal ribosomal entry site (IRES) and a nucleic acid encoding a self-cleaving 2A peptide (such as P2A, T2A, E2A, or F2A) linking the 3 end of second caTCR nucleic acid sequence to the 5 end of the first caTCR nucleic acid sequence, wherein the first caTCR nucleic acid sequence and the second caTCR nucleic acid sequence are transcribed as a single RNA under the control of the promoter. In some embodiments, the first and/or second promoters have the same sequence. In some embodiments, the first and/or second promoters have different sequences. In some embodiments, the first and/or second promoters are inducible. In some embodiments, the immune cell does not express the TCR subunits from which the TCR-TMs of the caTCR are derived. For example, in some embodiments, the immune cell is an T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and chains, or the immune cell is a T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and 0 chains. In some embodiments, the immune cell is modified to block or decrease the expression of one or both of its endogenous TCR subunits. For example, in some embodiments, the immune cell is an T cell modified to block or decrease the expression of the TCR and/or chains, or the immune cell is a T cell modified to block or decrease the expression of the TCR and/or chains. In some embodiments, the immune cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell. In some embodiments, the vector is a viral vector (such as a lentiviral vector) integrated into the host genome of the immune cell. It is to be appreciated that embodiments where any of the nucleic acid sequences are swapped are also contemplated, such as where the first or second caTCR nucleic acid sequence is swapped with the CSR nucleic acid sequence.

[0346] In some embodiments, there is provided a caTCR plus CSR immune cell (such as a T cell) expressing on its surface a caTCR according to any of the caTCRs described herein and a CSR according to any of the CSRs described herein, wherein the caTCR plus CSR immune cell comprises a vector comprising a) a first caTCR nucleic acid sequence encoding a first caTCR polypeptide chain of the caTCR; b) a second caTCR nucleic acid sequence encoding a second caTCR polypeptide chain of the caTCR; and c) a CSR nucleic acid sequence encoding a CSR polypeptide chain of the CSR, wherein the first and second caTCR nucleic acid sequences and the CSR nucleic acid sequence are under the control of a single promoter; wherein the first and second caTCR polypeptide chains are expressed from the first and second caTCR nucleic acid sequences to form the caTCR and the CSR polypeptide chain is expressed from the CSR nucleic acid sequence to form the CSR, and wherein the caTCR and CSR localize to the surface of the immune cell. In some embodiments, the promoter is operably linked to one of the nucleic acid sequences, which is linked to the other nucleic acid sequences by nucleic acid linkers selected, individually, from the group consisting of an internal ribosomal entry site (IRES) and a nucleic acid encoding a self-cleaving 2A peptide (such as P2A, T2A, E2A, or F2A), such that the first and second caTCR nucleic acid sequences and the CSR nucleic acid sequence are transcribed as a single RNA under the control of the promoter. In some embodiments, the promoter is inducible. In some embodiments, the immune cell does not express the TCR subunits from which the TCR-TMs of the caTCR are derived. For example, in some embodiments, the immune cell is an T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and chains, or the immune cell is a T cell and the TCR-TMs of the introduced caTCR comprise sequences derived from TCR and 0 chains. In some embodiments, the immune cell is modified to block or decrease the expression of one or both of its endogenous TCR subunits. For example, in some embodiments, the immune cell is an T cell modified to block or decrease the expression of the TCR and/or chains, or the immune cell is a T cell modified to block or decrease the expression of the TCR and/or chains. In some embodiments, the immune cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, a natural killer T cell, and a suppressor T cell. In some embodiments, the vector is a viral vector (such as a lentiviral vector) integrated into the host genome of the immune cell.

IV. Fc Variants

[0347] In some embodiments, anti-CD22 constructs described herein may comprise a variant Fc region, wherein the variant Fc region may comprise at least one amino acid modification relative to a reference Fc region (or parental Fc region or a wild-type Fc region). Amino acid modifications may be made in an Fc region to alter effector function and/or to increase serum stability of the construct. Construct comprising variant Fc regions may demonstrate an altered affinity for an Fc receptor (e.g., an FcR), provided that the variant Fc regions do not have a substitution at positions that make a direct contact with Fc receptor based on crystallographic and structural analysis of Fc-Fc receptor interactions such as those disclosed by Sondermann et al., 2000, Nature, 406: 267-273. Examples of positions within the Fc region that make a direct contact with an Fc receptor such as an FcR are amino acids 234-239 (hinge region), amino acids 265-269 (B/C loop), amino acids 297-299 (C/E loop), and amino acids 327-332 (F/G) loop. In some embodiments, constructs comprising variant Fc regions may comprise a modification of at least one residue that makes a direct contact with an FcR based on structural and crystallographic analysis.

[0348] Amino acid modifications in Fc regions to create variant Fc regions that, e.g., alter affinity for activating and/or inhibitory receptors, lead to improved effector function such as, e.g., Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC) and Complement Dependent Cytotoxicity (CDC), increase binding affinity for C1q, reduce or eliminate FcR binding, increase half-life are known in the art (see, e.g., U.S. Pat. Nos. 9,051,373, 9,040,041, 8,937,158, 8,883,973, 8,883,147, 8,858,937, 8,852,586, 8,809,503, 8,802,823, 8,802,820, 8,795,661, 8,753,629, 8,753,628, 8,735,547, 8,735,545, 8,734,791, 8,697,396, 8,546,543, 8,475,792, 8,399,618, 8,394,925, 8,388,955, 8,383,109, 8,367,805, 8,362,210, 8,338,574, 8,324,351, 8,318,907, 8,188,231, 8,124,731, 8,101,720, 8,093,359, 8,093,357, 8,088,376, 8,084,582, 8,039,592, 8,012,476, 7,799,900, 7,790,858, 7,785,791, 7,741,072, 7,704,497, 7,662,925, 7,416,727, 7,371,826, 7,364,731, 7,335,742, 7,332,581, 7,317,091, 7,297,775, 7,122,637, 7,083,784, 6,737,056, 6,538,124, 6,528,624 and 6,194,551).

[0349] In some embodiments, a variant Fc region may have different glycosylation patterns as compared to a parent Fc region (e.g., aglycosylated). In some embodiments, different glycosylation patterns may arise from expression in different cell lines, e.g., an engineered cell line.

[0350] Constructs described herein may comprise variant Fc regions that bind with a greater affinity to one or more FcRs. Such constructs preferably mediate effector function more effectively as discussed infra. In some embodiments, constructs described herein may comprise variant Fc regions that bind with a weaker affinity to one or more FcRs. Reduction or elimination of effector function may be desirable in certain cases, for example, in the case of constructs whose mechanism of action involves blocking or antagonism but not killing of the cells bearing a target antigen. In some embodiments, increased effector function may be directed to tumor cells and cells expressing foreign antigens.

V. Construct Production

[0351] Provided constructs or portions thereof, or nucleic acids encoding them, may be produced by any available means. Methods for construct production are well-known in the art. Technologies for generating antibodies (e.g., scFv antibodies, monoclonal antibodies, and/or polyclonal antibodies) are available in the art. It will be appreciated that a wide range of animal species can be used for the production of antisera, e.g., mouse, rat, rabbit, pig, cow, deer, sheep, goat, cat, dog, monkey, and chicken. The choice of animal may be decided upon the ease of manipulation, costs or the desired amount of sera, as would be known to one of skill in the art. It will be appreciated that antibodies can also be produced transgenically through the generation of a mammal or plant that is transgenic for the immunoglobulin heavy and light chain sequences of interest (e.g., a transgenic rodent transgenic for human immunoglobulin heavy and light chain genes). In connection with the transgenic production in mammals, antibodies can be produced in, and recovered from, the milk of goats, cows, or other mammals (see, e.g., U.S. Pat. Nos. 5,827,690, 5,756,687, 5,750,172, and 5,741,957; herein incorporated by reference in their entireties). Alternatively, antibodies may be made in chickens, producing IgY molecules (Schade et al., 1996, ALTEX 13(5): 80-85).

[0352] In some embodiments, antibodies suitable for the present invention are subhuman primate antibodies. For example, general techniques for raising therapeutically useful antibodies in baboons may be found, for example, in International Patent Application Publication No. 1991/11465 and in Losman et al., 1990, Int. J. Cancer 46: 310. In some embodiments, antibodies (e.g., monoclonal antibodies) may be prepared using hybridoma methods (Milstein and Cuello, 1983, Nature 305(5934): 537-40). In some embodiments, antibodies (e.g., monoclonal antibodies) may also be made by recombinant methods (see, e.g., U.S. Pat. No. 4,166,452).

[0353] Many of the difficulties associated with generating antibodies by B-cell immortalization can be overcome by engineering and expressing construct components in E.coli or yeast using phage display. To ensure the recovery of high affinity antibodies a combinatorial immunoglobulin library must typically contain a large repertoire size. A typical strategy utilizes mRNA obtained from lymphocytes or spleen cells of immunized mice to synthesize cDNA using reverse transcriptase. The heavy and light chain genes are amplified separately by PCR and ligated into phage cloning vectors. Two different libraries may be produced, one containing the heavy chain genes and one containing the light chain genes. The libraries can be nave or they can be semi-synthetic, i.e., with all amino acids (with the exception of cysteine) equally likely to be present at any given position in a CDR. Phage DNA is isolated from each library, and the heavy and light chain sequences are ligated together and packaged to form a combinatorial library. Each phage contains a random pair of heavy and light chain cDNAs and upon infection of E.coli directs the expression of the polypeptides in an anti-CD22 construct in infected cells. To identify a construct that recognizes the antigen of interest (e.g., CD22), the phage library is plated, and the construct molecules present in the plaques are transferred to filters. The filters are incubated with radioactively labeled antigen and then washed to remove excess unbound ligand. A radioactive spot on the autoradiogram identifies a plaque that contains a construct that binds the antigen. Alternatively, identification of a construct that recognizes the antigen of interest (e.g., CD22) may be achieved by iterative binding of phage to the antigen, which is bound to a solid support, for example, beads or mammalian cells followed by removal of non-bound phage and by elution of specifically bound phage. In such embodiments, antigens are first biotinylated for immobilization to, for example, streptavidin-conjugated Dynabeads M-280. The phage library is incubated with the cells, beads or other solid support and non-binding phage is removed by washing. Construct phage clones that bind the antigen of interest are selected and tested for further characterization.

[0354] Once selected, positive clones may be tested for their binding to the antigen of interest expressed on the surface of live cells by flow cytometry. Briefly, phage clones may be incubated with cells (e.g., engineered to express the antigen of interest, or those that naturally express the antigen) that either do or do not express the antigen. The cells may be washed and then labeled with a mouse anti-M13 coat protein monoclonal antibody. Cells may be washed again and labeled with a fluorescent-conjugated secondary antibody (e.g., FITC-goat (Fab).sub.2 anti-mouse IgG) prior to flow cytometry. Cloning and expression vectors that are useful for producing a human immunoglobulin phage library can be obtained, for example, from Stratagene Cloning Systems (La Jolla, Calif.).

[0355] A similar strategy may be employed to obtain high-affinity scFv clones. A library with a large repertoire may be constructed by isolating V-genes from non-immunized human donors using PCR primers corresponding to all known V.sub.H, V.sub. and V gene families. Following amplification, the V.sub. and V pools may be combined to form one pool. These fragments may be ligated into a phagemid vector. An scFv linker (e.g., (G.sub.4S)n) may be ligated into the phagemid upstream of the V.sub.L fragment (or upstream of the V.sub.H fragment as so desired). The V.sub.H and linker-V.sub.L fragments (or V.sub.L and linker-V.sub.H fragments) may be amplified and assembled on the J.sub.H region. The resulting V.sub.H-linker-V.sub.L (or V.sub.L-linker-V.sub.H) fragments may be ligated into a phagemid vector. The phagemid library may be panned using filters, as described above, or using immunotubes (Nunc; Maxisorp). Similar results may be achieved by constructing a combinatorial immunoglobulin library from lymphocytes or spleen cells of immunized rabbits and by expressing the scFv constructs in P. pastoris (see, e.g., Ridder et al., 1995, Biotechnology, 13: 255-260). Additionally, following isolation of appropriate scFv antibodies, higher binding affinities and slower dissociation rates may be obtained through affinity maturation processes such as mutagenesis and chain-shuffling (see, e.g., Jackson et al., 1998, Br. J. Cancer, 78: 181-188); Osbourn et al., 1996, Immunotechnology, 2: 181-196).

[0356] Human antibodies may be produced using various techniques, i.e., introducing human Ig genes into transgenic animals in which the endogenous Ig genes have been partially or completely inactivated can be exploited to synthesize human antibodies. In some embodiments, anti-CD22 human antibodies may be made by immunization of non-human animals engineered to make human antibodies in response to antigen challenge with human CD22.

[0357] Provided constructs may be also produced, for example, by utilizing a host cell system engineered to express an construct-encoding nucleic acid. Alternatively or additionally, provided constructs may be partially or fully prepared by chemical synthesis (e.g., using an automated peptide synthesizer or gene synthesis of construct-encoding nucleic acids). Constructs described herein may be expressed using any appropriate vector or expression cassette. A variety of vectors (e.g., viral vectors) and expression cassettes are known in the art and cells into which such vectors or expression cassettes may be introduced may be cultured as known in the art (e.g., using continuous or fed-batch culture systems). In some embodiments, cells may be genetically engineered; technologies for genetically engineering cells to express engineered polypeptides are well known in the art (see, e.g., Ausabel et al., eds., 1990, Current Protocols in Molecular Biology (Wiley, N.Y.)).

[0358] Constructs described herein may be purified, i.e., using filtration, centrifugation, and/or a variety of chromatographic technologies such as HPLC or affinity chromatography. In some embodiments, fragments of provided constructs are obtained by methods that include digestion with enzymes, such as pepsin or papain, and/or by cleavage of disulfide bonds by chemical reduction.

[0359] It will be appreciated that provided constructs may be engineered, produced, and/or purified in such a way as to improve characteristics and/or activity of the constructs. For example, improved characteristics include, but are not limited to, increased stability, improved binding affinity and/or avidity, increased binding specificity, increased production, decreased aggregation, decreased nonspecific binding, among others. In some embodiments, provided constructs may comprise one or more amino acid substitutions (e.g., in a framework region in the context of an immunoglobulin or fragment thereof (e.g., an scFv antibody)) that improve protein stability, antigen binding, expression level, or provides a site or location for conjugation of a therapeutic, diagnostic or detection agent.

VI. Therapeutic and Detection Agents

[0360] A therapeutic agent or a detection agent may be attached to an anti-CD22 construct described herein. Therapeutic agents may be any class of chemical entity including, for example, but not limited to, proteins, carbohydrates, lipids, nucleic acids, small organic molecules, non-biological polymers, metals, ions, radioisotopes, etc. In some embodiments, therapeutic agents for use in accordance with the present invention may have a biological activity relevant to the treatment of one or more symptoms or causes of cancer. In some embodiments, therapeutic agents for use in accordance with the present invention may have a biological activity relevant to modulation of the immune system and/or enhancement of T-cell mediated cytotoxicity. In some embodiments, therapeutic agents for use in accordance with the present invention have one or more other activities.

[0361] A detection agent may comprise any moiety that may be detected using an assay, for example due to its specific functional properties and/or chemical characteristics. Non-limiting examples of such agents include enzymes, radiolabels, haptens, fluorescent labels, phosphorescent molecules, chemiluminescent molecules, chromophores, luminescent molecules, photoaffinity molecules, colored particles or ligands, such as biotin.

[0362] Many detection agents are known in the art, as are systems for their attachment to constructs (see, for e.g., U.S. Pat. Nos. 5,021,236; 4,938,948; and 4,472,509). Examples of such detection agents include paramagnetic ions, radioactive isotopes, fluorochromes, NMR-detectable substances, X-ray imaging agents, among others. For example, in some embodiments, a paramagnetic ion is one or more of chromium (III), manganese (II), iron (III), iron (II), cobalt (II), nickel (II), copper (II), neodymium (III), samarium (III), ytterbium (III), gadolinium (III), vanadium (II), terbium (III), dysprosium (III), holmium (III), erbium (III), lanthanum (III), gold (III), lead (II), and/or bismuth (III).

[0363] The radioactive isotope may be one or more of actinium-225, astatine-211, bismuth-212, carbon-14, chromium-51, chlorine-36, cobalt-57, cobalt-58, copper-67, Europium-152, gallium-67, hydrogen-3, iodine-123, iodine-124, iodine-125, iodine-131, indium-111, iron-59, lead-212, lutetium-177, phosphorus-32, radium-223, radium-224, rhenium-186, rhenium-188, selenium-75, sulphur-35, technicium-99m, thorium-227, yttrium-90, and zirconium-89. Radioactively labeled constructs may be produced according to well-known technologies in the art.

[0364] A fluorescent label may be or may comprise one or more of Alexa 350, Alexa 430, AMCA, BODIPY 630/650, BODIPY 650/665, BODIPY-FL, BODIPY-R6G, BODIPY-TMR, BODIPY-TRX, Cascade Blue, Cy3, Cy5,6-FAM, Fluorescein Isothiocyanate, HEX, 6-JOE, Oregon Green 488, Oregon Green 500, Oregon Green 514, Pacific Blue, REG, Rhodamine Green, Rhodamine Red, Renographin, ROX, TAMRA, TET, Tetramethylrhodamine, and/or Texas Red, among others.

VII. Methods of Treatment

[0365] The anti-CD22 constructs and/or compositions of the invention can be administered to individuals (e.g., mammals such as humans) to treat a B-cell malignancy (e.g., a CD22.sup.+ B-cell malignancy), such as a B-cell lymphoma or a B-cell leukemia. B-cell malginancies also include various types of cancers as described further herein. Examples of B-Cell malignancies include, without limitation, acute lymphoblastic leukemia (ALL), Hodgkin's lymphoma, non-Hodgkin's lymphoma, B cell chronic lymphocytic leukemia (CLL), multiple myeloma, follicular lymphoma, mantle cell lymphoma, pro-lymphocytic leukemia, hairy cell leukemia, common acute lymphocytic leukemia, and null-acute lymphoblastic leukemia. The present invention provides method of treating a B-cell malignancy (e.g., a CD22.sup.+ B-cell malignancy) in an individual comprising administering to the individual an effective amount of a pharmaceutical composition comprising an anti-CD22 construct described herein (e.g., an anti-CD22 scFv), in which the anti-CD22 construct binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof).

[0366] Cancers

[0367] The anti-CD22 constructs and cells expressing anti-CD22 constructs including anti-CD22 CAR and anti-CD22 caTCR in some embodiments can be useful for treating B-cell related cancer. Cancers that may be treated using any of the methods described herein include tumors that are not vascularized, or not yet substantially vascularized, as well as vascularized tumors. The cancers may comprise non-solid tumors (such as hematological tumors, for example, leukemias and lymphomas) or may comprise solid tumors. Types of cancers to be treated with the anti-CD22 constructs and anti-CD22 CAR cells of the invention include, but are not limited to, carcinoma, blastoma, sarcoma, melanoma, neuroendocrine tumors, and glioma, and certain leukemia or lymphoid malignancies, benign and malignant tumors, and malignancies e.g., sarcomas, carcinomas, melanomas, and gliomas. Adult tumors/cancers and pediatric tumors/cancers are also included.

[0368] Solid tumors contemplated for treatment by any of the methods described herein include CNS tumors, such as glioma (e.g., brainstem glioma and mixed gliomas), glioblastoma (also known as glioblastoma multiforme), astrocytoma (such as high-grade astrocytoma), pediatric glioma or glioblastoma (such as pediatric high-grade glioma (HGG) and diffuse intrinsic pontine glioma (DIPG)), CNS lymphoma, germinoma, medulloblastoma, Schwannoma craniopharyogioma, ependymoma, pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma, menangioma, neuroblastoma, retinoblastoma and brain metastases.

[0369] In some embodiments, the B-cell related cancer is pediatric glioma. In some embodiments, the pediatric glioma is a low-grade glioma. In some embodiments, the pediatric glioma is a high-grade glioma (HGG). In some embodiments, the pediatric glioma is glioblastoma multiforme. In some embodiments, the pediatric glioma is diffuse intrinsic pontine glioma (DIPG). In some embodiments, the DIPG is grade II. In some embodiments, the DIPG is grade III. In some embodiments, the DIPG is grade IV.

[0370] Additional solid tumors contemplated for treatment include fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma (such as clear-cell chondrosarcoma), chondroblastoma, osteosarcoma, and other sarcomas, synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, lymphoid malignancy, pancreatic cancer, breast cancer, lung cancers, ovarian cancer, prostate cancer, hepatocellular carcinoma, squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweat gland carcinoma, medullary thyroid carcinoma, papillary thyroid carcinoma, pheochromocytomas sebaceous gland carcinoma, papillary carcinoma, papillary adenocarcinomas, medullary carcinoma, bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma, Wilms' tumor, cervical cancer (e.g., cervical carcinoma and pre-invasive cervical dysplasia), cancer of the anus, anal canal, or anorectum, vaginal cancer, cancer of the vulva (e.g., squamous cell carcinoma, intraepithelial carcinoma, adenocarcinoma, and fibrosarcoma), penile cancer, oropharyngeal cancer, head cancers (e.g., squamous cell carcinoma), neck cancers (e.g., squamous cell carcinoma), testicular cancer (e.g., seminoma, teratoma, embryonal carcinoma, teratocarcinoma, choriocarcinoma, sarcoma, Leydig cell tumor, fibroma, fibroadenoma, adenomatoid tumors, and lipoma), bladder carcinoma, melanoma, cancer of the uterus (e.g., endometrial carcinoma), and urothelial cancers (e.g., squamous cell carcinoma, transitional cell carcinoma, adenocarcinoma, ureter cancer, and urinary bladder cancer).

[0371] Hematologic cancers contemplated for treatment by any of the methods described herein include leukemias, including acute leukemias (such as acute lymphocytic leukemia, acute myelocytic leukemia, acute myelogenous leukemia and myeloblastic, promyelocytic, myelomonocytic, monocytic and erythroleukemia), chronic leukemias (such as chronic myelocytic (granulocytic) leukemia, chronic myelogenous leukemia, and chronic lymphocytic leukemia), polycythemia vera, lymphoma, Hodgkin's disease, non-Hodgkin's lymphoma (indolent and high grade forms), multiple myeloma, Waldenstrom's macroglobulinemia, heavy chain disease, myelodysplastic syndrome, hairy cell leukemia and myelodysplasia.

[0372] Cancer treatments can be evaluated, for example, by tumor regression, tumor weight or size shrinkage, time to progression, duration of survival, progression free survival, overall response rate, duration of response, quality of life, protein expression and/or activity. Approaches to determining efficacy of the therapy can be employed, including for example, measurement of response through radiological imaging.

[0373] In some embodiments, the anti-CD22 construct used in methods of treating a B-cell malignancy (e.g., a CD22.sup.+ B-cell malignancy) comprises an antibody moiety that comprises i) a light chain variable region comprising one or more of LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, and ii) a heavy chain variable region comprising one or more of HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively. In some embodiments, the antibody moiety comprises LC-CDR1, LC-CDR2, LC-CDR3, HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 206-211, respectively. In other embodiments, the anti-CD22 construct used in methods of treating a B-cell malignancy comprises an antibody moiety that comprises i) a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 206-208, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 212, and ii) a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209-211, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 213, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker. In some embodiments, the anti-CD22 construct is a full-length antibody. In some embodiments, the anti-CD22 construct is a multispecific antibody (e.g., a bispecific antibody). In some embodiments, the anti-CD22 construct is a CAR or caTCR. In some embodiments, the anti-CD22 construct is an immunoconjugate comprising the antibody moiety described above and an effector molecule. The effector molecule may be a therapeutic agent (e.g., a drug, a toxin, a radioisotope, a protein, a peptide, or a nucleic acid) or a label. In some embodiments, the therapeutic agent is a drug or a toxin. In some embodiments, the composition further comprises a cell (such as an effector cell) associated with the anti-CD22 construct.

[0374] In some embodiments, the anti-CD22 construct used in methods of treating a B-cell malignancy (e.g., a CD22.sup.+ B-cell malignancy) comprises an antibody moiety that comprises i) a light chain variable region comprising one or more of LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, and ii) a heavy chain variable region comprising one or more of HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively. In some embodiments, the antibody moiety comprises LC-CDR1, LC-CDR2, LC-CDR3, HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 214-216, 209, 210, and 217, respectively. In other embodiments, the anti-CD22 construct used in methods of treating a B-cell malignancy comprises an antibody moiety that comprises i) a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 214-216, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 218, and ii) a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209, 210, and 217, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 219, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker. In some embodiments, the anti-CD22 cosntruct is a full-length antibody. In some embodiments, the anti-CD22 cosntruct is a multispecific antibody (e.g., a bispecific antibody). In some embodiments, the anti-CD22 cosntruct is a CAR or caTCR. In some embodiments, the anti-CD22 cosntruct is an immunoconjugate comprising the antibody moiety described above and an effector molecule. The effector molecule may be a therapeutic agent (e.g., a drug, a toxin, a radioisotope, a protein, a peptide, or a nucleic acid) or a label. In some embodiments, the therapeutic agent is a drug or a toxin. In some embodiments, the composition further comprises a cell (such as an effector cell) associated with the anti-CD22 cosntruct.

[0375] In some embodiments of any of the methods for treating a B-cell malignancy (e.g., a CD22.sup.+ B-cell malignancy described above, the anti-CD22 construct is conjugated to a cell (such as an immune cell, e.g., a T cell) prior to being administered to the individual. Thus, for example, there is provided a method of treating a B-cell malignancy (e.g., a CD22.sup.+ B-cell malignancy) in an individual comprising a) conjugating an anti-CD22 construct described herein or an antibody moiety theerof to a cell (such as an immune cell, e.g., a T cell) to form an anti-CD22 construct/cell conjugate, and b) administering to the individual an effective amount of a composition comprising the anti-CD22 construct/cell conjugate. In some embodiments, the cell is derived from the individual. In some embodiments, the cell is not derived from the individual. In some embodiments, the anti-CD22 construct is conjugated to the cell by covalent linkage to a molecule on the surface of the cell. In some embodiments, the anti-CD22 construct is conjugated to the cell by non-covalent linkage to a molecule on the surface of the cell. In some embodiments, the anti-CD22 construct is conjugated to the cell by insertion of a portion of the anti-CD22 construct into the outer membrane of the cell.

[0376] Treatments can be evaluated, for example, by tumor regression, tumor weight or size shrinkage, time to progression, duration of survival, progression free survival, overall response rate, duration of response, quality of life, protein expression and/or activity. Approaches to determining efficacy of the therapy can be employed, including for example, measurement of response through radiological imaging.

[0377] In some embodiments, the efficacy of treatment may be measured as the percentage tumor growth inhibition (% TGI), which may be calculated using the equation 100-(T/C100), where T is the mean relative tumor volume of the treated tumor, and C is the mean relative tumor volume of a non-treated tumor. In some embodiments, the % TGI is about 2%, about 4%, about 6, about 8%, 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94% , about 95%, or more than 95%.

VIII. Anti-CD22 Construct Effector Cell Therapy

[0378] The present application also provides methods of using an anti-CD22 construct (such as an anti-CD22 CAR, anti-CD22 caTCR, or anti-CD22 CSR) to redirect the specificity of an effector cell (such as a primary T cell) to a CD22.sup.+ cell in a B-cell malignancy. Thus, the present invention also provides a method of stimulating an effector cell-mediated response (such as a T cell-mediated immune response) to a target cell population or tissue comprising CD22.sup.+ cells in a mammal, comprising the step of administering to the mammal an effector cell (such as a T cell) that expresses an anti-CD22 CAR or anti-CD22 caTCR. In some embodiments, stimulating an immune cell refers to eliciting an effector cell-mediated response (such as a T cell-mediated immune response), which is different from activating an immune cell. In some embodiments, an anti-CD22 CSR can stimulate an immune cell (e.g., a T cell), but does not activate the immune cell.

[0379] Anti-CD22 construct effector cells (such as anti-CD22 CAR T cells or anti-CD22 caTCR T cells) expressing the anti-CD22 construct can be infused to a recipient in need thereof. The infused cell is able to kill CD22.sup.+ cells in the recipient. In some embodiments, unlike antibody therapies, anti-CD22 construct effector cells (such as T cells) are able to replicate in vivo resulting in long-term persistence that can lead to sustained tumor control.

[0380] In some embodiments, the anti-CD22 construct effector cells are anti-CD22 CAR T cells or anti-CD22 caTCR T cells that can undergo robust in vivo T cell expansion and can persist for an extended amount of time. In some embodiments, the anti-CD22 CAR T cells or anti-CD22 caTCR T cells of the invention develop into specific memory T cells that can be reactivated to inhibit any additional tumor formation or growth.

[0381] The anti-CD22 construct T cells (such as anti-CD22 CAR T cells or anti-CD22 caTCR T cells) of the invention may also serve as a type of vaccine for ex vivo immunization and/or in vivo therapy in a mammal. In some embodiments, the mammal is a human.

[0382] With respect to ex vivo immunization, at least one of the following occurs in vitro prior to administering the cell into a mammal: i) expansion of the cells, ii) introducing a nucleic acid encoding an anti-CD22 CAR or anti-CD22 caTCR to the cells, and/or iii) cryopreservation of the cells. Ex vivo procedures are well-known in the art. Briefly, cells are isolated from a mammal (preferably a human) and genetically modified (i.e., transduced or transfected in vitro) with a vector expressing an anti-CD22 CAR or anti-CD22 caTCR disclosed herein. The anti-CD22 CAR cell or anti-CD22 caTCR cell can be administered to a mammalian recipient to provide a therapeutic benefit. The mammalian recipient may be a human and the anti-CD22 CAR or anti-CD22 caTCR cell can be autologous with respect to the recipient. Alternatively, the cells can be allogeneic, syngeneic or xenogeneic with respect to the recipient. The procedure for ex vivo expansion of hematopoietic stem and progenitor cells is described in U.S. Pat. No. 5,199,942, incorporated herein by reference, can be applied to the cells of the present invention. Other suitable methods are known in the art, therefore the present invention is not limited to any particular method of ex vivo expansion of the cells. Briefly, ex vivo culture and expansion of T cells comprises: (1) collecting T cells from peripheral blood mononuclear cells (PBMC); and (2) expanding such cells ex vivo. In addition to the cellular growth factors described in U.S. Pat. No. 5,199,942, other factors such as flt3-L, IL-1, IL-3 and c-kit ligand, can be used for culturing and expansion of the cells.

[0383] In addition to using a cell-based vaccine in terms of ex vivo immunization, the present invention also provides compositions and methods for in vivo immunization to elicit an immune response directed against an antigen in a patient. The anti-CD22 construct effector cells (such as anti-CD22 CAR T cells or anti-CD22 caTCR T cells) of the present invention may be administered either alone, or as a pharmaceutical composition in combination with diluents and/or with other components such as IL-2 or other cytokines or cell populations. Briefly, pharmaceutical compositions of the present invention may comprise anti-CD22 construct effector cells (such as T cells), in combination with one or more pharmaceutically or physiologically acceptable carriers, diluents or excipients. Such compositions may comprise buffers such as neutral buffered saline, phosphate buffered saline and the like; carbohydrates such as glucose, mannose, sucrose or dextrans, mannitol; proteins; polypeptides or amino acids such as glycine; antioxidants; chelating agents such as EDTA or glutathione; adjuvants (e.g., aluminum hydroxide); and preservatives. In some embodiments, anti-CD22 construct effector cell (such as T cell) compositions are formulated for administration by intravenous, intrathecal, intracranial, intracerebral, or intracerebroventricular route.

[0384] The precise amount of the anti-CD22 construct effector cell (such as anti-CD22 CAR T cell or anti-CD22 caTCR T cell) compositions of the present invention to be administered can be determined by a physician with consideration of individual differences in age, weight, tumor size, extent of infection or metastasis, and condition of the patient (subject). In some embodiments, a pharmaceutical composition comprising the anti-CD22 construct effector cells (such as T cells) is administered at a dosage of about 10.sup.4 to about 10.sup.9 cells/kg body weight, such any of about 10.sup.4 to about 10.sup.5, about 10.sup.5 to about 10.sup.6, about 10.sup.6 to about 10.sup.7, about 10.sup.7 to about 10.sup.8, or about 10.sup.8 to about 10.sup.9 cells/kg body weight, including all integer values within those ranges. Anti-CD22 construct effect cell (such as T cell) compositions may also be administered multiple times at these dosages. The cells can be administered by using infusion techniques that are commonly known in immunotherapy (see, e.g., Rosenberg et al., New Eng. J. of Med. 319: 1676, 1988). The optimal dosage and treatment regimen for a particular patient can readily be determined by one skilled in the art of medicine by monitoring the patient for signs of disease and adjusting the treatment accordingly.

[0385] In some embodiments, it may be desired to administer activated anti-CD22 construct T cells (such as anti-CD22 CAR T cells or anti-CD22 caTCR T cells) to a subject and then subsequently redraw blood (or have an apheresis performed), activate T cells therefrom according to the present invention, and reinfuse the patient with these activated and expanded T cells. This process can be carried out multiple times every few weeks. In some embodiments, T cells can be activated from blood draws of from 10 cc to 400 cc. In some embodiments, T cells are activated from blood draws of 20 cc, 30 cc, 40 cc, 50 cc, 60 cc, 70 cc, 80 cc, 90 cc, or 100 cc.

[0386] The administration of the anti-CD22 construct effector cells (such as anti-CD22 CAR T cells or anti-CD22 caTCR T cells) may be carried out in any convenient manner, including by injection, ingestion, transfusion, implantation or transplantation. The compositions described herein may be administered to a patient subcutaneously, intradermally, intratumorally, intranodally, intramedullary, intramuscularly, intrathecally, intracranially, intracerebrally, intracerebroventricularly, by intravenous (i.v.) injection, or intraperitoneally. In some embodiments, the anti-CD22 construct effector cell (such as T cell) compositions of the present invention are administered to a patient by intradermal or subcutaneous injection. In some embodiments, the anti-CD22 construct effector cell (such as T cell) compositions of the present invention are administered by i.v. injection. In some embodiments, the anti-CD22 construct effector cell (such as T cell) compositions of the present invention are administered by intrathecal injection. In some embodiments, the anti-CD22 construct effector cell (such as T cell) compositions of the present invention are administered by intracranial injection. In some embodiments, the anti-CD22 construct effector cell (such as T cell) compositions of the present invention are administered by intracerebral injection. In some embodiments, the anti-CD22 construct effector cell (such as T cell) compositions of the present invention are administered by intracerebroventricular injection. The compositions of anti-CD22 construct effector cells (such as T cells) may be injected directly into a tumor, lymph node, or site of infection.

[0387] Thus, for example, in some embodiments, there is provided a method of treating a B-cell malignancy (e.g., a CD22.sup.+ B-cell malignancy) in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-CD22 CAR comprising a) an extracellular domain comprising an anti-CD22 construct described herein or an antibody moiety thereof that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3t intracellular signaling sequence and a CD28 and/or 4-1BB intracellular signaling sequence. In some embodiments, the anti-CD22 antibody moiety comprises i) a light chain variable region comprising one or more of LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, and ii) a heavy chain variable region comprising one or more of HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively. In some embodiments, the antibody moiety comprises LC-CDR1, LC-CDR2, LC-CDR3, HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 206-211, respectively. In other embodiments, the anti-CD22 antibody moiety comprises i) a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 206-208, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 212, and ii) a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209-211, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 213, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker.

[0388] Thus, for example, in some embodiments, there is provided a method of treating a B-cell malignancy (e.g., a CD22.sup.+ B-cell malignancy) in an individual comprising administering to the individual an effective amount of a composition comprising an effector cell (such as a T cell) expressing an anti-CD22 CAR comprising a) an extracellular domain comprising an anti-CD22 antibody moiety that specifically binds to an extracellular region of CD22 or a portion thereof (e.g., SEQ ID NO: 205 or a portion thereof), b) a transmembrane domain, and c) an intracellular signaling domain comprising a CD3 intracellular signaling sequence and a CD28 and/or 4-1BB intracellular signaling sequence. In some embodiments, the anti-CD22 antibody moiety comprises i) a light chain variable region comprising one or more of LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, and ii) a heavy chain variable region comprising one or more of HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively. In some embodiments, the antibody moiety comprises LC-CDR1, LC-CDR2, LC-CDR3, HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 214-216, 209, 210, and 217, respectively. In other embodiments, the anti-CD22 antibody moiety comprises i) a light chain variable region having LC-CDR1, LC-CDR2, and LC-CDR3 of the sequences of SEQ ID NOS: 214-216, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 218, and ii) a heavy chain variable region having HC-CDR1, HC-CDR2, and HC-CDR3 of the sequences of SEQ ID NOS: 209, 210, and 217, respectively, and a sequence that has at least 90% (e.g., at least 92%, 94%, 96%, 98%, or 99%) identity to the sequence of SEQ ID NO: 219, in which the light chain variable region and the heavy chain variable region are joined to each other via a linker.

IX. Methods of Diagnosis and Imaging Using Anti-CD22 Constructs

[0389] Labeled anti-CD22 antibody moieties and derivatives and analogs thereof, which specifically bind to a CD22 on the surface of a cell, can be used for diagnostic purposes to detect, diagnose, or monitor a B-cell malignancy (e.g., a B-cell related cancer or a CD22.sup.+ B-cell malignancy). For example, the anti-CD22 antibody moieties of the invention can be used in in situ, in vivo, ex vivo, and in vitro diagnostic assays or imaging assays.

[0390] Additional embodiments of the invention include methods of diagnosing a B-cell malignancy (e.g., a B-cell related cancer or a CD22.sup.+ B-cell malignancy) in an individual (e.g., a mammal such as a human). The methods comprise detecting CD22-presenting cells in the individual. In some embodiments, the B-cell malignancy is a B-cell lymphoma or a B-cell leukemia. In some embodiments, there is provided a method of diagnosing a B-cell malignancy (e.g., a B-cell related cancer or a CD22.sup.+ B-cell malignancy) in an individual (e.g., a mammal, such as a human) comprising (a) administering an effective amount of a labeled anti-CD22 antibody moiety according to any of the embodiments described above to the individual; and (b) determining the level of the label in the individual, such that a level of the label above a threshold level indicates that the individual has the B-cell malignancy. The threshold level can be determined by various methods, including, for example, by detecting the label according to the method of diagnosing described above in a first set of individuals that have the B-cell malignancy and a second set of individuals that do not have the B-cell malignancy, and setting the threshold to a level that allows for discrimination between the first and second sets. In some embodiments, the threshold level is zero, and the method comprises determining the presence or absence of the label in the individual. In some embodiments, the method further comprises waiting for a time interval following the administering of step (a) to permit the labeled anti-CD22 antibody moiety to preferentially concentrate at sites in the individual where the CD22 is expressed (and for unbound labeled anti-CD22 antibody moiety to be cleared). In some embodiments, the method further comprises subtracting a background level of the label. Background level can be determined by various methods, including, for example, by detecting the label in the individual prior to administration of the labeled anti-CD22 antibody moiety, or by detecting the label according to the method of diagnosing described above in an individual that does not have the B-cell malignancy.

[0391] Anti-CD22 antibody moieties of the invention can be used to assay levels of CD22-presenting cell in a biological sample using methods known to those of skill in the art. Suitable antibody labels are known in the art and include enzyme labels, such as, glucose oxidase; radioisotopes, such as iodine (131I, 125I, 123I, 121I), carbon (14C), sulfur (35S), tritium (3H), indium (115mIn, 113mIn, 112In, 111In), technetium (99Tc, 99mTc), thallium (201Ti), gallium (68Ga, 67Ga), palladium (103Pd), molybdenum (99Mo), xenon (133Xe), fluorine (18F), samarium (153Sm), lutetium (177Lu), gadolinium (159Gd), promethium (149Pm), lanthanum (140La), ytterbium (175Yb) , holmium (166Ho), yttrium (90Y), scandium (47Sc), rhenium (186Re, 188Re), praseodymium (142Pr), rhodium (105Rh), and ruthenium (97Ru); luminol; fluorescent labels, such as fluorescein and rhodamine; and biotin.

[0392] Techniques known in the art may be applied to labeled anti-CD22 antibody moieties of the invention. Such techniques include, but are not limited to, the use of bifunctional conjugating agents (see e.g., U.S. Pat. Nos. 5,756,065; 5,714,631; 5,696,239; 5,652,361; 5,505,931; 5,489,425; 5,435,990; 5,428,139; 5,342,604; 5,274,119; 4,994,560; and 5,808,003). Aside from the above assays, various in vivo and ex vivo assays are available to the skilled practitioner. For example, one can expose cells within the body of the subject to an anti-CD22 antibody moiety which is optionally labeled with a detectable label, e.g., a radioactive isotope, and binding of the anti-CD22 antibody moiety to the cells can be evaluated, e.g., by external scanning for radioactivity or by analyzing a sample (e.g., a biopsy or other biological sample) derived from a subject previously exposed to the anti-CD22 antibody moiety.

X. Pharmaceutical Compositions

[0393] Also provided herein are compositions (such as pharmaceutical compositions, also referred to herein as formulations) comprising an anti-CD22 construct described herein, a nucleic acid encoding one or more polypeptides contained in an anti-CD22 construct described herein, an expression cassette comprising the nucleic acid, or a host cell expressing an anti-CD22 construct. In some embodiments, the composition further comprises a cell (such as an effector cell, e.g., a T cell) associated with the anti-CD22 construct. In some embodiments, there is provided a pharmaceutical composition comprising an anti-CD22 construct and a pharmaceutically acceptable carrier. In some embodiments, the pharmaceutical composition further comprises a cell (such as an effector cell, e.g., a T cell) associated with the anti-CD22 construct.

[0394] Suitable formulations of the anti-CD22 constructs are obtained by mixing an anti-CD22 construct having the desired degree of purity with optional pharmaceutically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions. Acceptable carriers, excipients, or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid and methionine; preservatives (such as octadecyldimethylbenzyl ammonium chloride; hexamethonium chloride; benzalkonium chloride, benzethonium chloride; phenol, butyl or benzyl alcohol; alkyl parabens such as methyl or propylparaben; catechol; resorcinol; cyclohexanol; 3-pentanol; and m-cresol); low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as olyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, histidine, arginine, or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugars such as sucrose, mannitol, trehalose or sorbitol; salt-forming counter-ions such as sodium; metal complexes (e.g. Zn-protein complexes); and/or non-ionic surfactants such as TWEEN, PLURONICS or polyethylene glycol (PEG). Exemplary formulations are described in WO98/56418, expressly incorporated herein by reference. Lyophilized formulations adapted for subcutaneous administration are described in WO97/04801. Such lyophilized formulations may be reconstituted with a suitable diluent to a high protein concentration and the reconstituted formulation may be administered subcutaneously to the individual to be treated herein. Lipofectins or liposomes can be used to deliver the anti-CD22 antibodies s of this invention into cells.

[0395] The formulation herein may also contain one or more active compounds in addition to the anti-CD22 construct as necessary for the particular indication being treated, preferably those with complementary activities that do not adversely affect each other. For example, it may be desirable to further provide an anti-neoplastic agent, a growth inhibitory agent, a cytotoxic agent, or a chemotherapeutic agent in addition to the anti-CD22 construct. Such molecules are suitably present in combination in amounts that are effective for the purpose intended. The effective amount of such other agents depends on the amount of anti-CD22 construct present in the formulation, the type of disease or disorder or treatment, and other factors discussed above. These are generally used in the same dosages and with administration routes as described herein or about from 1 to 99% of the heretofore employed dosages.

[0396] The anti-CD22 antibodies may also be entrapped in microcapsules prepared, for example, by coacervation techniques or by interfacial polymerization, for example, hydroxymethylcellulose or gelatin-microcapsules and poly-(methylmethacylate) microcapsules, respectively, in colloidal drug delivery systems (for example, liposomes, albumin microspheres, microemulsions, nano-particles and nanocapsules) or in macroemulsions. Such techniques are disclosed in Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980). Sustained-release preparations may be prepared.

[0397] Sustained-release preparations of the anti-CD22 constructs can be prepared. Suitable examples of sustained-release preparations include semipermeable matrices of solid hydrophobic polymers containing the construct (or fragment thereof), which matrices are in the form of shaped articles, e.g., films, or microcapsules. Examples of sustained-release matrices include polyesters, hydrogels (for example, poly(2-hydroxyethyl-methacrylate), or poly(vinylalcohol)), polylactides (U.S. Pat. No. 3,773,919), copolymers of L-glutamic acid and ethyl-L-glutamate, non-degradable ethylene-vinyl acetate, degradable lactic acid-glycolic acid copolymers such as the LUPRON DEPOT TM (injectable microspheres composed of lactic acid-glycolic acid copolymer and leuprolide acetate), and poly-D-()-3-hydroxybutyric acid. While polymers such as ethylene-vinyl acetate and lactic acid-glycolic acid enable release of molecules for over 100 days, certain hydro gels release proteins for shorter time periods. When encapsulated constructs remain in the body for a long time, they can denature or aggregate as a result of exposure to moisture at 37 C., resulting in a loss of biological activity and possible changes in immunogenicity. Rational strategies can be devised for stabilization of anti-CD22 constructs depending on the mechanism involved. For example, if the aggregation mechanism is discovered to be intermolecular SS bond formation through thio-disulfide interchange, stabilization can be achieved by modifying sulfhydryl residues, lyophilizing from acidic solutions, controlling moisture content, using appropriate additives, and developing specific polymer matrix compositions.

[0398] In some embodiments, the anti-CD22 construct is formulated in a buffer comprising a citrate, NaCl, acetate, succinate, glycine, polysorbate 80 (Tween 80), or any combination of the foregoing. In some embodiments, the anti-CD22 construct is formulated in a buffer comprising about 100 mM to about 150 mM glycine. In some embodiments, the anti-CD22 construct is formulated in a buffer comprising about 50 mM to about 100 mM NaCl. In some embodiments, the anti-CD22 construct is formulated in a buffer comprising about 10 mM to about 50 mM acetate. In some embodiments, the anti-CD22 construct is formulated in a buffer comprising about 10 mM to about 50 mM succinate. In some embodiments, the anti-CD22 construct is formulated in a buffer comprising about 0.005% to about 0.02% polysorbate 80. In some embodiments, the anti-CD22 construct is formulated in a buffer having a pH between about 5.1 and 5.6. In some embodiments, the anti-CD22 construct is formulated in a buffer comprising 10 mM citrate, 100 mM NaCl, 100 mM glycine, and 0.01% polysorbate 80, wherein the formulation is at pH 5.5.

[0399] The formulations to be used for in vivo administration must be sterile. This is readily accomplished by, e.g., filtration through sterile filtration membranes.

XI. Dosage and Administration

[0400] The dose of the anti-CD22 construct compositions administered to an individual (such as a human) may vary with the particular composition, the mode of administration, and the type of disease being treated. In some embodiments, the amount of the anti-CD22 construct composition is sufficient to result in a complete response in the individual. In some embodiments, the amount of the anti-CD22 construct composition is sufficient to result in a partial response in the individual. In some embodiments, the amount of the anti-CD22 construct composition administered (for example when administered alone) is sufficient to produce an overall response rate of more than about any of 2%, 4%, 6%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 64%, 65%, 70%, 75%, 80%, 85%, or 90% among a population of individuals treated with the anti-CD22 construct composition. Responses of an individual to the treatment of the methods described herein can be determined, for example, based on the percentage tumor growth inhibition (% TGI).

[0401] In some embodiments, the amount of the composition is sufficient to prolong overall survival of the individual. In some embodiments, the amount of the composition (for example when administered along) is sufficient to produce clinical benefit of more than about any of 2%, 4%, 6%, 8%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 60%, 70%, or 77% among a population of individuals treated with the anti-CD22 construct composition.

[0402] In some embodiments, the amount of the composition is an amount sufficient to decrease the size of a tumor, decrease the number of cancer cells (e.g., CD22.sup.+ cells), or decrease the growth rate of a tumor by at least about any of 2%, 4%, 6%, 8%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95% or 100% compared to the corresponding tumor size, number of cancer cells, or tumor growth rate in the same subject prior to treatment or compared to the corresponding activity in other subjects not receiving the treatment. Standard methods can be used to measure the magnitude of this effect, such as in vitro assays with purified enzyme, cell-based assays, animal models, or human testing.

[0403] In some embodiments, the amount of the anti-CD22 construct in the composition is below the level that induces a toxicological effect (i.e., an effect above a clinically acceptable level of toxicity) or is at a level where a potential side effect can be controlled or tolerated when the composition is administered to the individual. In some embodiments, the amount of the composition is close to a maximum tolerated dose (MTD) of the composition following the same dosing regimen. In some embodiments, the amount of the composition is more than about any of 80%, 90%, 95%, or 98% of the MTD. In some embodiments, the amount of an anti-CD22 construct in the composition is included in a range of about 0.001 g to about 1000 g. In some embodiments of any of the above aspects, the effective amount of an anti-CD22 construct in the composition is in the range of about 0.1 g/kg to about 100 mg/kg of total body weight.

[0404] The anti-CD22 construct compositions can be administered to an individual (such as human) via various routes, including, for example, intravenous, intra-arterial, intraperitoneal, intrapulmonary, oral, nasal, inhalation, intravesicular, intramuscular, intra-tracheal, subcutaneous, intraocular, intrathecal, intracranial, intracerebral, intracerebroventricular, transmucosal, and transdermal. In some embodiments, sustained continuous release formulation of the composition may be used. In some embodiments, the composition is administered intravenously. In some embodiments, the composition is administered intraarterially. In some embodiments, the composition is administered intraperitoneally. In some embodiments, the composition is administered intrathecally. In some embodiments, the composition is administered intracranially. In some embodiments, the composition is administered intracerebrally. In some embodiments, the composition is administered intracerebroventricularly. In some embodiments, the composition is administered nasally.

EXAMPLES

Example 1

Generation and Selection of Human Anti-CD22 Constructs

[0405] This example demonstrates the production of human constructs specific for human CD22. In particular, this example demonstrates the production of human single chain variable fragments (scFvs) that specifically bind human CD22 in native format. Human constructs described herein were developed using naive or semi-synthetic human phage libraries developed from normal donors and/or diseases donors and selected based on high specificity for CD22 via panning on cell surface expressed human CD22 in its native conformation. Thus, such human constructs may serve as a valuable source for construction of, among other things, full-length IgG, multispecific antibodies, and chimeric antigen receptors that may otherwise be deleted from repertoires found in nature.

[0406] Briefly, an exemplary outline for the development of anti-human CD22 constructs is set forth in Table 2. The process started with identification of human CD22-specific and biologically active constructs from the E-ALPHA phage library. A collection of human scFv antibody phage display libraries (diversity=1010.sup.10) constructed at Eureka Therapeutics, named as E-ALPHA phage libraries, was used for the selection of human constructs specific to human CD22. E-ALPHA phage libraries included naive libraries consisting of fully naive human heavy and light chain repertoires, and semi-synthetic libraries containing fully naive human light chain repertoires and semi-synthetic heavy chain with completely randomized heavy chain CDR3 regions. The naive antibody repertoires were cloned from PBMCs and spleens of healthy donors or from PBMCs of disease donors. The scFv libraries were used in panning against human CD22 positive cells including Raji (a lymphoma cell line naturally expressing CD22), Jurkat cells expressing full-length CD22, and Jurkat cells expressing domains 5-7 of CD22. The expression of full-length CD22 in Jurkat cells was confirmed by anti-CD22 staining using APC mouse anti-human CD22 antibody (Biolegend, Cat. No. 363505), which only binds to full-length CD22 as in FIGS. 1A-1E. Domains 5-7 of CD22 was conjugated to green fluorescent protein (GFP) and its expression in Jurkat cells was confirmed by FITC signal. Jurkat cells without CD22 expression was used as a negative control. For cell panning, Jurkat cells expressing full-length CD22 or domains 5-7 of CD22, or Raji cells were first mixed with human scFv phage libraries. After extended washing with PBS, cells with bound scFv antibody phage were spun down. The bound clones were then eluted and used to infect E.coli XL1-Blue cells. The phage clones were expressed in bacteria and purified. The panning was performed for three to four rounds to enrich for scFv phage clones that specifically bound the extracellular region of human CD22.

TABLE-US-00008 TABLE 2 Stage Methodology Primary panning with ALPHA cell panning phage library Flow cytometry screening of phage clones Clone Characterization Target cancer cell killing Human B cell binding Clone Characterization Flow Cytometry Target cancer cell killing

[0407] Phage clones selected by cell panning were tested for cell-surface human CD22 binding directly. Out of the 1080 clones screened by phage display, 2 clones were identified as cell-surface CD22 binding clones as confirmed by flow cytometry. As shown in FIGS. 2A and 2B, clones 1 and 2 demonstrated specific binding to CD22.sup.+ Raji cells and Jurkat cells expressing full-length CD22 or domains 5-7 of CD22. Moreover, clones 1 and 2 were also demonstrated to bind specifically to NALM-6 cells (a leukemia cell line naturally expressing CD22) as shown in FIGS. 3A and 3B.

Example 2

Characterization of T Cells Expressing Anti-Human CD22 Chimeric Antigen Receptors (CAR-T)

[0408] This example describes the construction of chimeric antigen receptors (CARs) using anti-human CD22 antibodies. In particular, this example specifically describes the construction of CARs that include an antigen-binding site of an anti-CD22 antibody and are expressed on the surface of T cells. Further, the CARs expressed by T cells were employed in cytotoxicity assays against human lymphoma cell lines. Thus, the present example illustrates that, in some embodiments, using CARs that include an antigen binding site from human anti-CD22 antibodies described herein is useful for killing target cells that express human CD22 (e.g., lymphoma).

[0409] In vitro cytotoxicity of human CD22 transduced T cells. Lenti-viruses containing human CD22 specific chimeric antigen receptors (CARs) were produced by transfection of 293T cells with CAR vectors. In some embodimetns, human T cells were used for transduction after one-day stimulation with CD3/CD28 beads (Dynabeads, Invitrogen) in the presence of IL-2 at 100 U/mL. Concentrated lenti-viruses were applied to T cells in Retronectin (Takara) coated 6-well plates for 72 hours. Functional assessment of transduced T cells (CD22/CAR-T cells) is performed using a LDH Cytotoxicity Assay.

[0410] Moreover, in a similar experiment, human CD22 CAR-Ts (described above) were tested using a large panel of CD22 positive and negative cancer cell lines. Briefly, primary T cells were mock-transduced (Mock) or transduced with selected CAR encoding anti-CD22 antibodies. Transduced T cells were analyzed by FACS using an antibody to detect a tag (e.g., a myc tag) in the extracellular domain of the CAR constructs.

[0411] In another experiment, cytokine release profile and target cell lysis of activated human CD22 CAR-T cells was determined. Mock-transduced T cells (mock) or selected anti-CD22 CAR-Ts were co-incubated with target cells (FIG. 4A: K562, K562+CD22, and K562+M18+CD22; FIG. 4B: ASPC1, Jurkat, IM9, MCF7, HepG2, Hela, SK-Hep1, and SKOV3; FIG. 4C: K562, K562+CD22, LnCap, Colo205, and NALM6; FIG. 5A: K562 and K562+CD22; FIG. 5B: ASPC1, Jurkat, IM9, MCF7, HepG2, Hela, SK-Hep1, and SKOV3) in a 2 to 1 ratio for 16 hours. Release of IFN- into the media after in vitro killing was measured using the BioPlex 200 system (Bio-Rad) with the Bio-plex Pro Human Cytokine 8-plex Assays (BioRad). Cytokine concentrations were determined using a known standard curve, after subtracting release from media, target cell alone, and clone transduced T cell alone. The results of the IFN- release assays are shown in FIGS. 4A, 4B, and 4C, which illustrate that anti-CD22 CAR (clone 2) displayed specific and potent IFN- release against CD22.sup.+ target cells. FIGS. 5A and 5B further show that anti-CD22 CAR (clone 2) demonstrated specific killing against K562+CD22 target cells and no killing against K562 cells. FIG. 5A also shows that anti-CD22 CAR (clone 2) demonstrated superior killing activity compared to m971 CAR cells (m971 is an antibody that targets the membrane proximal region of CD22).

Example 3

Competition Assay

[0412] A flow cytometry competition assay was carried out to determine if m971 competes with anti-CD22 CAR (clone 2). m971 is an antibody that targets the membrane proximal region of CD22. Target cells K562+CD22 were pre-incubated with blocking antibody (m971 or anti-CD22 CAR (clone 1)) at 10 g/mL, 5 g/mL, 1 g/mL, 0.1 g/mL, 0.01 g/mL, and 0.001 g/mL for 30 minutes. After pre-incubation, 1 g/mL of detecting antibody (anti-CD22 CAR (clone 2) conjugated to biotin) was added directly to the sample without washing and incubated for another 30 minutes. After blocking, FACS was used for detection and analysis. The percent binding was determined from the MFI (mean fluorescence intensity) and samples were normalized against the isotype control. FIG. 6 shows that m971 does not compete with anti-CD22 CAR (clone 2), which binds to a different epitope. FIG. 6 also shows that anti-CD22 CAR (clone 2) and anti-CD22 CAR (clone 1) compete with each other, indicating that they likely bind the same epitope or an overlapping epitope. Anti-CD22 CAR (clone 1) appeared to have higher affinity than anti-CD22 CAR (clone 2) which appeared to have higher affinity than m971.

Example 4

Generation of Bispecific Constructs Using Human Anti-CD22 Antibodies

[0413] This example describes the construction of multispecific antibodies using human scFvs specific for human CD22. In particular, this example specifically describes the construction of bispecific antibodies having a first antigen-binding site that binds human CD22 in native format (cell-surface expressed) and a second antigen-binding site. In some embodiments, the second antigen-binding site is a protein expressed on T cells (e.g., CD3 on T cells). Thus, the present example illustrates that, in some embodiments, using multispecific antibodies that contain anti-CD22 antibody moieties as described herein, T cells are directed to kill target cells that express human CD22.

[0414] Bispecific antibodies are generated using scFv sequences of the human CD22-specific phage clones (e.g., clones 1 and 2). The bispecific antibodies are constructed using a single-chain format comprising the V.sub.L-V.sub.H scFv sequence of a human CD22-specific phage clone at the N-terminal end and an anti-human CD3monoclonal scFv at the C-terminal end (e.g., see Brischwein et al., Mol. Immunol. 43: 1129, 2006). The DNA fragments encoding the human CD22 scFv and the anti-human CD3scFv are synthesized and subcloned into a mammalian expression vector, e.g., pQD-T (Eureka Therapeutics, Inc.) using standard recombinant DNA technology. A hexhistamine tag is inserted at the C-terminal end for purification and detection. Mammalian cells, e.g., HEK293 cells, are transfected with the bispecific antibody expression vector and cultured for bispecific antibody production. Bispecific antibodies may subsequently be purified from cell supernatants using, e.g., HisTrap HP column. Cell culture is clarified and loaded onto the column with low imidazole concentration (e.g., 20 mM), and then an isocratic high imidazole concentration elution buffer (e.g., 500 mM) is used to elute bound bispecific antibodies. Molecular weights of purified human CD22 bispecific antibodies are measured under non-reducing conditions by gel electrophoresis.

Example 5

Characterization of Human CD22 Bispecific Antibodies

[0415] This example describes the characterization of the binding profile of bispecific antibodies.

[0416] Binding to recombinant human CD22 ECD-Fc fusion protein. Phage clones identified as specific binders to cell surface human CD22, e.g., clones 1 and 2, are tested for binding to recombinant human CD22 ECD-Fc fusion protein in solution. In some embodiments, biotinylated human CD22 ECD-Fc fusion proteins are loaded onto a streptavidin biosensor. After washing off excess antigen, bispecific antibodies are tested in PBS buffer for association and dissociation.

[0417] Binding to primary human B-cells. Human B-cells are tested for anti-CD22 antibody binding by, for example, co-staining human PBMCs with PerCP-conjugated anti-human CD20 antibody, antibodies targeting the second antigen-binding site (e.g., APC-labeled anti-human CD3 antibodies), and anti-CD22 bispecific antibodies. After a round of brief washing with PBS buffer, FITC-labeled anti-His tag antibody is added to the mixture as the secondary antibody for detection of the bispecific antibody. For the flow cytometry assay, human B-cells are gated by positive CD20 staining and negative CD3 staining. The anti-CD22 bispecific antibodies are evaluated for their ability to recognize human CD22 expressed on these CD2O+CD3- cells.

[0418] T-cell Killing assay. Tumor cytotoxicity is assayed using, e.g., LDH Cytotoxicity Assay (Promega). Human T cells (AllCells) or Ficoll-purified cells from whole blood (Blood Centers of the Pacific) are activated and expanded with, e.g., CD3/CD28 Dynabeads (Invitrogen). Activated T cells are cultured and maintained in, e.g., RPMI1640 medium with 10% FBS plus 100 U/mL IL-2. Activated T cells are used after a few days, e.g., 7-14 days, post-activation. FACS analysis is used to confirm T cell activation. Activated T cells and target cells are co-cultured at a 5: 1 ratio with bispecific antibodies. Cytotoxicity is determined by measuring LDH activity in culture supernatants.

Example 6

Affinity Maturation of Anti-Human CD22 Antibodies

[0419] This example describes the affinity maturation of anti-human CD22 antibodies. In particular, generation of a series of antibody variants is performed by incorporation of random mutations into selected anti-human CD22 antibodies (clones 1 and 2) followed by screening and characterization of the antibody variants.

[0420] Generation of variant phage libraries. DNA encoding anti-human CD22 scFvs is subjected to random mutagenesis using, e.g., GeneMorph II Random Mutagenesis kit (Agilent Technologies). After mutagenesis, DNA sequences are cloned into an scFv-expressing phagemid vector to build variant antibody phage libraries. Mutation libraries are built for each anti-human CD22 specific clone separately. Individual phage clones from enriched phage panning pools (e.g., variant clones) are tested for enhanced binding to cell-surface human CD22 compared to their respective parental clones. Further, a competition cell-binding assay is performed to compare the binding affinities of the variant clones to those of the parental clones.

Example 7

Generation and Characterization of Bispecific Antibodies Based on Anti-Human CD22 Variant Clones

[0421] This example describes the construction of multispecific antibodies using variant human scFvs specific for human CD22. In particular, this example specifically describes the construction of bispecific antibodies having a first antigen-binding site that binds human CD22 in native format (cell-surface expressed) and a second antigen-binding site. In some embodiments, the second antigen-binding site may bind CD3 on T cells. Thus, the present example illustrates that, in some embodiments, using multispecific antibodies that contain antibody moieties as described herein, T cells are directed to kill target cells that express human CD22.

[0422] Generation of variant clone bispecific antibodies. Bispecific antibodies derived from affinity-improved, variant antibody clones are generated as described in Example 2.

[0423] Binding affinity determination for variant bispecific antibodies. Relative binding affinity of the variant clones as compared to parental antibodies is determined through antibody titration flow cytometry using, e.g., human CD22.sup.+ cancer cells. Bispecific antibody clones, at serially diluted concentrations, are mixed with Raji cells. Antibody EC.sub.50 and apparent KD are calculated based on flow cytometry binding signals.

[0424] T-cell Killing assay. Tumor cytotoxicity is determined using, e.g., LDH Cytotoxicity Assay (Promega). Human T cells are activated and expanded with, e.g., CD3/CD28 Dynabeads (Invitrogen). Activated T cells are cultured and maintained in, e.g., RPMI1640 medium with 10% FBS plus 100 U/mL IL-2. Activated T cells are used a few days, e.g., 7-14 days, post-activation. FACS analysis is used to confirm T cell activation. Activated T cells and target cells are co-cultured at a 5: 1 ratio with bispecific antibodies. Cytotoxicity is determined by measuring LDH activity in culture supernatants.

Example 8

Characterization of T Cells Expressing Anti-Human CD22 Chimeric Antigen Receptors (CAR-T)

[0425] In another similar experiment, CAR-Ts generated from selected human antibodies described herein and a non-human (e.g., murine) antibody are tested using a panel of CD22 positive and negative cancer cell lines as described above. Briefly, primary T cells are mock-transduced (Mock) or transduced with selected CAR encoding anti-human CD22 scFvs described herein (e.g., clone 1 or clone 2) or CAR encoding anti-human CD22 scFv which has variable region sequences from an anti-human CD22 murine antibody. Transduced T cells are analyzed by FACs as described above.

[0426] In vivo efficacy of CD22 CAR-T cells in human lymphoma xenografts. The in vivo antitumor activity of an exemplary CAR-T cell is tested in a CD22.sup.+ human lymphoma xenograft model in NOD SCID gamma (NSG) mice. Cell line Raji-luc-GFP is derived from the CD22.sup.+ Burkitt lymphoma cell line, Raji, after stable transfection with dual reporter genes encoding both firefly luciferase (luc) and green fluorescent protein, which results in cells that are able to be traced in vivo using bioluminescent imaging. NSG mice are purchased from Jackson Laboratories (Bar Harbor, Me. USA 04609). Raji-luc-GFP cells are re-suspended in PBS and implanted intravenously (i.v.) into NSG mice through tail vein at 110.sup.6 cells/100 L/mouse. Five days post-implantation, animals are imaged using Xenogen IVIS imaging system for assessment of tumor burden. Mice are randomized into three groups: (i) no treatment, (ii) mock (non-transduced activated human T cells from the same donor of CAR-T cells), and (iii) clone CAR-T. Animals are treated i.v. with Mock or clone CAR-T cells immediately after randomization at a dose of, e.g., 10.sup.7 T cells per mouse (comprising 6-810.sup.6 CAR+ T cells per dose for group (iii)), once every two weeks for 3 doses. Animals are closely monitored after dosing. Bioluminescent imaging using Xenogen IVIS system is taken once a week. Animals with the following conditions are euthanized and recorded as conditional death: (i) Body weight loss more than 25% initial body weight and (ii) limb paralysis that affects mouse movement.

[0427] Post tumor implantation, mice from the anti-CD22 CAR clone-transduced T cell treatment group are re-challenged by i.v. implantation with Raji lymphoma cells to determine if the anti-CD22-CAR-transduced T cells would persist and maintain the capacity to respond to antigen (CD22). Nave NSG mice (i.e., mice not implanted with Raji lymphoma cells or previously treated with T cells) are implanted with Raji lymphoma cells one-day post injection of mock-transduced T cells as a control. Such mock-transduced T cells may mimic a condition of low level circulating T cells in the mice prior to implantation of Raji lymphoma cells. Tumor burden in each group is measured by luciferase activity.

[0428] In another similar experiment, in vivo antitumor activity of an exemplary CAR-T cell is tested in a CD22.sup.+ human leukemia xenograft model in NSG mice (NALM). NALM-6-luc-GFP cells are derived from the CD22.sup.+ acute lymphoblastic leukemia cell line NALM-6 after stable transfection with dual reporter genes encoding both firefly luciferase (luc) and green fluorescent protein (GFP), resulting in cells traceable in vivo using bioluminescent imaging.

[0429] Briefly, NALM-6-luc-GFP is cultured in RPMI Medium+10% FBS at 37 C. in a humidified atmosphere with 5% CO.sub.2. NSG mice are purchased from Jackson Laboratories (Bar Harbor, ME USA 04609) and acclimated prior to experimentation. NALM-6-luc-GFP cells are re-suspended in phosphate-buffered saline (PBS) and intravenously (i.v.) implanted into female NSG mice at, e.g., 510.sup.5 cells/100 L/mouse via tail vein injection. Post implantation, animals are imaged using Xenogen IVIS imaging system for tumor burden assessment. NSG mice are randomized into three groups: (i) vehicle (PBS); (ii) mock-transduced human T cells; and (iii) clone anti-CD22 CAR-transduced T cells. Animals are closely monitored after implantation and dosing with T cells or vehicle as described above.

Example 9

Generation and Characterization of T Cells Expressing Monospecific Anti-Human CD22 Chimeric Antibody-T Cell Receptors (caTCR)

[0430] This example describes the generation and characterization of T cells expressing various monospecific caTCR constructs that comprise anti-human CD22 Fabs described herein. Some of the caTCR constructs comprise both an anti-CD22 Fab and an anti-CD22 scFv described herein. Some of the caTCR-T cells further express a chimeric signaling receptor (CSR).

[0431] Generation of T cells expressing anti-human CD22-caTCR. Briefly, primary T cells were mock-transduced (Mock) or transduced with selected caTCR encoding nucleic acids. In particular, nucleic acids encoding the following caTCR constructs or caTCR+CSR construct combinations were generated and used in the transduction:

[0432] Construct combination 1 (SEQ ID NO: 1): anti-CD19-caTCR +anti-CD19-CSR

[0433] Construct 2 (SEQ ID NO: 2): anti-CD22-caTCR

[0434] Construct combination 3 (SEQ ID NO: 3): anti-CD22-caTCR +anti-CD19-CSR

[0435] Construct 4 (SEQ ID NO: 4): anti-CD22-scFv-anti-CD22-caTCR (also referred to as bivalent-monospecific anti-CD22-caTCR)

[0436] Construct combination 5 (SEQ ID NO: 5): anti-CD22-scFv-anti-CD22-caTCR +anti-CD19-CSR

[0437] Construct combination 6 (SEQ ID NO: 6): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR.

[0438] In construct combinations 1, 3, 5, and 6, the CSR comprises a truncated CD28 (SEQ ID NO: 157), which comprises CD28 transmembrane region sequence and intracellular signaling sequences.

[0439] In addition, nucleic acids encoding the following construct combinations are generated and transduced into primary T cells:

[0440] Construct combination 7 (SEQ ID NO: 7): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR-with CD8 TM sequence and a 4-1BB IC signaling sequence (SEQ ID NO: 173)

[0441] Construct combination 8 (SEQ ID NO: 8): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR-with truncated 4-1BB (SEQ ID NO: 159)

[0442] Construct combination 9 (SEQ ID NO: 9): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR-with CD8 TM sequence and a CD27 IC signaling sequence (SEQ ID NO: 167)

[0443] Construct combination 10 (SEQ ID NO: 10): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR-with truncated CD27 (SEQ ID NO: 161)

[0444] Construct combination 11 (SEQ ID NO: 11): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR-with CD8 TM sequence and a CD30 IC signaling sequence (SEQ ID NO: 169)

[0445] Construct combination 12 (SEQ ID NO: 12): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR-with truncated CD30 (SEQ ID NO: 163)

[0446] Construct combination 13 (SEQ ID NO: 13): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR-with CD8 TM sequence and a OX40 IC signaling sequence (SEQ ID NO: 171)

[0447] Construct combination 14 (SEQ ID NO: 14): anti-CD22-scFv-anti-CD22-caTCR +anti-CD22-CSR-with truncated OX40 (SEQ ID NO: 165).

[0448] Further, nucleic acids encoding the construct combinations 5-14 (SEQ ID NOS: 5-14) but with the myc tag (SEQ ID NO: 194) removed are also generated and transduced into primary T cells.

[0449] In the constructs and construct combinations disclosed in this Example, each CSR and the co-expressed caTCR are translated as a single polypeptide initially, and then get cleaved into separate polypeptides. In other embodiments, co-expressed CSR and caTCR can be constructed as separately translated polypeptides encoded on the same nucleic acid or even encoded on different nucleic acids.

[0450] Characterization of T cells expressing anti-human CD22 caTCR. Primary T cells were transduced with nucleic acides encoding construct combinations 1, 3, 5, and 6 (SEQ ID NOS: 1, 3, 5, and 6, respectively), constructs 2 and 4 (SEQ ID NOS: 2 and 4, respectively), or mock transduced (with no nucleic acids). Transduction efficiency was determined by cell surface staining, with Fab as the marker for caTCR expression and myc tag as the marker for CSR expression. The results indicated that caTCR.sup.+ cell percentages and CSR.sup.+ cell percentages were about the same in the same T cell samples transduced with caTCR-CSR-encoding nucleci acids. All caTCR-transduced T-cells were matched at (normalized to) approximately 46% caTCR receptor positive by mixing with mock T-cells, and caTCR.sup.+ cells were used as the effector cells.

[0451] In vitro killing. CD80/86 negative NALM6-luc-GFP cells and CD80/CD86 positive Raji-luc-GFP cells (leukemia and lymphoma cells expressing CD22 and CD19) were used as target cells in separate experiments for T-cell stimulation at an effector-to-target ratio of 1: 1 and incubated with the caTCR T cells for overnight (about 16 h). Specific T-cell lysis was measured after the overnight incubation using the Cytox 96 Non-radioactive Cytotoxicity Assay (Promega). Live target cells were counted, and killing percentage was calculated as the difference between the percentage of the remaining live target cells in each group compared to that of the group with target alone (no T cells). The result are shown in FIG. 7. The caTCR T cells expressing any one of the six constructs and construct combinations (SEQ ID NOS: 1-6) had very high killing efficacy in NALM6 cells. They also had sigfinicant killing efficacy in Raji cells, although not as high as in NALM6 at the tested effector-to-target ratio of 1: 1.

[0452] Similar in vitro killing experiments are carried out using the NALM6-luc-GFP cells and Raji-luc-GFP cells as the target cells and primary T cells transduced with nucleic acides encoding construct combinations 7-14 as the effector cells.

[0453] Cytokine secretion. In addition to measuring live target cell numbers, the concentration of IFN- released into the supernatant of the in vitro killing reactions was measured as another indicator of cell killing with a Human IFN- ELISA MAX kit from Biolegend. The results are shown in FIG. 8. The caTCR T cells expressing any one of the six constructs and construct combinations had high IFN- release levels from Raji cells, especially construct 4 (anti-CD22-scFv-anti-CD22-caTCR (also referred to as bivalent-monospecific anti-CD22-caTCR)). Most of the anti-CD22-caTCR T cell groups also had high IFN- release levels from NALM6 cells.

[0454] Similarly, the concentration of IFN- released into the supernatant of the in vitro killing reactions from NALM6 cells and Raji cells with T cells expressing construct combinations 7-14 is measured with the same method.

[0455] Target cell re-challenge. T cells expressing any one of construct combinations 3, 5, and 6 (SEQ ID NOS: 1, 3, 5, and 6, respectively) and constructs 2 and 4 (SEQ ID NOS: 2 and 4, respectively) were initially co-cultured with 50,000 NALM6-luc-GFP or Raji-luc-GFP target cells at an effector-to-target ratio of 1: 1. Every 7 days 100,000 of new target cells were added to the same co-culture to re-challenge (or engage) the T cells. Remaining target cells and T cells in the co-culture were counted twice a week using flow analysis to evaluate the killing activity of T cells. The results with NALM6 as the target cells are shown in FIG. 9A (target cell numbers) and FIG. 9B (total T cell number), while the results with Raji as the target cells are shown in FIG. 10A (target cell numbers) and FIG. 10B (total T cell number). Anti-CD22-caTCR T cells expressing any one of the five constructs and construct combinations (SEQ ID NOS: 2-6) maintained high killing efficacy in NALM6 cells up to at least three weeks and after three or more rounds of target engagments (e.g., at E3D7 and E4D3). For NALM6 target cells, T cells expressing construct combination 5 (anti-CD22-scFv-anti-CD22-caTCR +anti-CD19-CSR) persisted for the longest time among the five constructs and construct combinations. Anti-CD22-caTCR T cells expressing any one of the five constructs and construct combinations also had sigfinicant killing efficacy in Raji cells up to about five weeks and after a total of five rounds of target engagements.

[0456] Similar target cell re-challenge experiments are carried out using the NALM6-luc-GFP cells and Raji-luc-GFP cells as the target cells and primary T cells transduced with nucleic acids encoding each of construct combinations 1 and 7-14 as the effector cells.

[0457] The results of the experiments described in this Example show that T cells transduced to express anti-CD22-caTCR successfully killed target cancer cells expressing CD22. Such cells also persisted for at least weeks and maintained their target cell killing capability.

Example 10

Generation and Characterization of T Cells Expressing Bispecific or Trispecific Anti-Human CD22-caTCR

[0458] This example describes the generation and characterization of T cells expressing various bispecific or trispecific caTCR constructs that comprise anti-human CD22 Fabs and one or two additional antibody variable region fragments targeting non-CD22 antigens as described herein. Some of the caTCR-T cells further express a CSR.

[0459] Generation of T cells expressing various caTCR including bispecific or trispecific anti-human CD22-caTCR. Briefly, primary T cells were mock-transduced (Mock) or transduced with selected caTCR encoding nucleic acids. In particular, nucleic acids encoding the following constructs 15-21 (SEQ ID NOS: 15-21, respectively) were generated and used in the transduction.

[0460] In vitro killing. Following a similar protocol as described in Example 9, CD80/86 negative NALM6-luc-GFP cells, CD80/CD86 positive Raji-luc-GFP cells (leukemia and lymphoma cells expressing CD22 and CD19), and K562 cells were used as target cells in separate experiments for T-cell stimulation at an effector-to-target ratio of 1: 1 and incubated with the caTCR T cells for overnight (about 16 h). Specific T-cell lysis was measured after the overnight incubation using the Cytox 96 Non-radioactive Cytotoxicity Assay (Promega). Live target cells were counted, and killing percentage was calculated as the difference between the percentage of the remaining live target cells in each group compared to that of the group with target alone (no T cells). The result are shown in FIG. 11 and FIGS. 12A-12F. The caTCR T cells expressing any one of the bispecific constructs (constructs 15-21 (SEQ ID NOS: 15-21, respectively)) or monospecific constructs (constructs 2, 22, and 28 (SEQ ID NOS: 2, 22, and 28, respectively)) had very high killing efficacy in the cells.

[0461] Similar in vitro killing experiments are carried out following the same experimental protocol using nucleic acides encoding construct combination 23 (SEQ ID NO: 23), constructs 24-27 (SEQ ID NOS: 24-27, respectively), and construct combinations 29-75 (SEQ ID NOS: 29-75, respectively).

[0462] Target cell re-challenge. Following a similar protocol as described in Example 9, T cells expressing construct combinations 1 and 23 were initially co-cultured with 50,000 NALM6-luc-GFP target cells at an effector-to-target ratio of 1: 1. Every 7 days 100,000 of new target cells were added to the same co-culture to re-challenge (or engage) the T cells. Remaining target cells and T cells in the co-culture were counted twice a week using flow analysis to evaluate the killing activity of T cells. The results with NALM6 as the target cells are shown in FIG. 13A and FIG. 13B.

[0463] Similar target cell re-challenge experiments are carried out following the same experimental protocol using nucleic acids encoding each of constructs and construct combinations 15-21, 24-27, and 29-75 (SEQ ID NOS: 15-21, 24-27, and 29-75, respectively).

Example 11

Generation and Characterization of T Cells Expressing other Construct Combinations

[0464] This example describes the generation and characterization of T cells expressing various other caTCR-CSR construct combinations that comprise anti-human CD22 Fabs and one or two additional antibody variable region fragments targeting non-CD22 antigens as described herein. Some of the caTCR-T cells further express a CSR.

[0465] Following a similar protocol as described above, primary T cells are mock-transduced (Mock) or transduced with nucleic acids encoding the following construct combinations:

[0466] (1) a construct combination that comprises a caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 77 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96;

[0467] (2) a construct combination that comprises a caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 77 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112;

[0468] (3) a construct combination that comprises a caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 77 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128;

[0469] (4) a construct combination that comprises a caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 78 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 81-96;

[0470] (5) a construct combination that comprises a caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 78 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 97-112; and

[0471] (6) a construct combination that comprises a caTCR and a CSR, in which the caTCR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of SEQ ID NO: 78 and the CSR has a sequence that has at least 90% (e.g., 92%, 94%, 96%, 98%, 99%, or 100%) identity to the sequence of any one of SEQ ID NOS: 113-128.

[0472] In some embodiments, an anti-CD22 construct combination comprises a caTCR that has a light chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 212 and a heavy chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 213.

[0473] In some embodiments, an anti-CD22 construct combination comprises a caTCR that has a light chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 218 and a heavy chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 219.

[0474] In some embodiments, an anti-CD22 construct combination comprises a CSR that has a light chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 212 and a heavy chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 213.

[0475] In some embodiments, an anti-CD22 construct combination comprises a CSR that has a light chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 218 and a heavy chain variable region having a sequence that has at least 90% identity (e.g., at least 92%, 94%, 96%, 98%, or 99% identity) to the sequence of SEQ ID NO: 219.

[0476] Nucleic acids encoding any of the anti-CD22 construct combinations described in any of the above embodiments are also transduced into primary T cells.

[0477] Following a similar protocol as described above, any of the anti-CD22 construct combinations described in any of the above embodiments are also tested in in vitro killing and target cell re-challenge experiments.

Example 12

In Vivo Efficacy Study of T Cells Transduced with Anti-CD22 Constructs or Construct Combinations

[0478] The in vivo anti-tumor activity of T cells expressing one or more of anti-CD22 constructs and construct combinations described herein is tested in a human CD19.sup.+ NALM-6 pre-B Acute Lymphoblastic Leukemia (ALL) model. Luciferase-expressing NALM-6 cells are implanted intravenously (i.v.) into NOD SCID gamma (NSG) immune-compromised mice and tumor burden is assessed by measuring tumor-derived bioluminescence. Six days post tumor implantation, mice are randomized based on total bioluminescent flux into treatment groups: (1) i.v. injection of 510.sup.6 un-transduced donor-matched (Mock) T cells, (2) i.v. injection of 210.sup.6 T cells expressing an anti-CD22 caTCR construct only (caTCR T cell) and (3) i.v. injection of 210.sup.6 T cells expressing both anti-CD19 caTCR and anti-CD19 CSR (caTCR CSR T cell; n=6 mice/group). Health effects resulting from T cell infusions in mice are assessed by monitoring their general appearance, body weight, and other clinical signs of adverse response (including hypothermia, labored respiration, and hind-limb paralysis/weakness).

[0479] To determine the level of cytokine release in vivo, key cytokines, including those related to clinical cytokine release syndrome, are analyzed 24 hours after the NALM-6 tumor-bearing mice are administered with anti-CD22 CAR-T cells or caTCR CSR T cells. Cytokine levels are quantified with Luminex Magpix technology using BioRad Bio-Plex kits.

EXEMPLARY EMBODIMENTS

[0480] Exemplary embodiments provided in accordance with the presently disclosed subject matter include, but are not limited to, the embodiments and the following embodiments: [0481] 1. An anti-CD22 construct comprising an antibody moiety that specifically binds to CD22, wherein the antibody moiety comprises:

[0482] (a) a light chain variable region (V.sub.L ) comprising a light chain complementarity determining region (LC-CDR) 1, an LC-CDR2, and an LC-CDR3 of the light chain variable region of SEQ ID NO: 218 or 212; and

[0483] (b) a heavy chain variable region (V.sub.H ) comprising a heavy chain complementarity determining region (HC-CDR) 1, an HC-CDR2, and an HC-CDR3 of the heavy chain variable region of SEQ ID NO: 219 or 213. [0484] 2. The anti-CD22 construct of embodiment 1, wherein the antibody moiety comprises the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the light chain variable region of SEQ ID NO: 218. [0485] 3. The anti-CD22 construct of embodiment 1, wherein the antibody moiety comprises the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the light chain variable region of SEQ ID NO: 212. [0486] 4. The anti-CD22 construct of embodiment 1 or 2, wherein the antibody moiety comprises:

[0487] (a) the light chain variable region (VL) comprising the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the light chain variable region of SEQ ID NO: 218; and (b) a heavy chain variable region (VH) comprising the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the heavy chain variable region of SEQ ID NO: 219. [0488] 5. The anti-CD22 construct of embodiment 1 or 3, wherein the antibody moiety comprises:

[0489] (a) the light chain variable region (VL) comprising the LC-CDR1, the LC-CDR2, and the LC-CDR3 of the light chain variable region of SEQ ID NO: 212; and

[0490] (b) a heavy chain variable region (VH) comprising the HC-CDR1, the HC-CDR2, and the HC-CDR3 of the heavy chain variable region of SEQ ID NO: 213. [0491] 6. The anti-CD22 construct of embodiment 1, wherein the antibody moiety comprises one or more of:

[0492] the LC-CDR1 having a sequence of HDIRNY (SEQ ID NO: 214),

[0493] the LC-CDR2 having a sequence of AAS (SEQ ID NO: 215),

[0494] the LC-CDR3 having a sequence of QQYDGLPLT (SEQ ID NO: 216),

[0495] the HC-CDR1 having a sequence of GFTFSNYA (SEQ ID NO: 209),

[0496] the HC-CDR2 having a sequence of ISGSGGST (SEQ ID NO: 210), and

[0497] the HC-CDR3 having a sequence of ARYGSAAWMDS (SEQ ID NO: 217). [0498] 7. The anti-CD22 construct of embodiment 6, wherein the antibody moiety comprises the sequences of SEQ ID NOS: 209, 210, and 214-217. [0499] 8. The anti-CD22 construct of embodiment 6 or 7, wherein the light chain variable region has a sequence having at least 90% identity to the sequence of DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTGV PSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKR (SEQ ID NO: 218). [0500] 9. The anti-CD22 construct of any one of embodiments 6 to 8, wherein the heavy chain variable region has a sequence having at least 90% identity to the sequence of QVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVS SISGSGG STYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQG TLVTVSS (SEQ ID NO: 219). [0501] 10. An anti-CD22 construct comprising a light chain variable region and a heavy chain variable region, wherein

[0502] the light chain variable region has a sequence having at least 90% identity to the sequence of DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTGV PSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKR (SEQ ID NO: 218), and

[0503] the heavy chain variable region has a sequence having at least 90% identity to the sequence of QVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGG STYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQG TLVTVSS (SEQ ID NO: 219). [0504] 11. The anti-CD22 construct of embodiment 10, wherein the light chain variable region comprises the sequence of SEQ ID NO: 218, and the heavy chain variable region comprises the sequence of SEQ ID NO: 219. [0505] 12. The anti-CD22 construct of embodiment 1, wherein the antibody moiety comprises one or more of:

[0506] the LC-CDR1 having a sequence of SSNIGNNY (SEQ ID NO: 206),

[0507] the LC-CDR2 having a sequence of ENN (SEQ ID NO: 207),

[0508] the LC-CDR3 having a sequence of GTWDSSLSAGAV (SEQ ID NO: 208),

[0509] the HC-CDR1 having a sequence of GFTFSNYA (SEQ ID NO: 209),

[0510] the HC-CDR2 having a sequence of ISGSGGST (SEQ ID NO: 210), and

[0511] the HC-CDR3 having a sequence of ARPYYDD (SEQ ID NO: 211). [0512] 13. The anti-CD22 construct of embodiment 12, wherein the antibody moiety comprises the sequences of SEQ ID NOS: 206-211. [0513] 14. The anti-CD22 construct of embodiment 12 or 13, wherein the light chain variable region has a sequence having at least 90% identity to the sequence of QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLIYENNKRPSG IPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLSAGAVFGGGTKLTVLG (SEQ ID NO: 212). [0514] 15. The anti-CD22 construct of any one of embodiments 12 or 13, wherein the heavy chain variable region has a sequence having at least 90% identity to the sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSAISGSGG STYYAD SVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARPYYDDWGQGTLVT VSS (SEQ ID NO: 213). [0515] 16. An anti-CD22 construct comprising a heavy chain variable region and a light chain variable region and, wherein

[0516] the light chain variable region has a sequence having at least 90% identity to the sequence of QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLIYENNKRPSG IPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLSAGAVFGGGTKLTVLG (SEQ ID NO: 212), and

[0517] the heavy chain variable region has a sequence having at least 90% identity to the sequence of EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSAISGSGG STYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARPYYDDWGQGTLVT VSS (SEQ ID NO: 213). [0518] 17. The anti-CD22 construct of embodiment 16, wherein the light chain variable region comprises the sequence of SEQ ID NO: 212, and the heavy chain variable region comprises the sequence of SEQ ID NO: 213. [0519] 18. An anti-CD22 construct comprising an antibody moiety that competes with the anti-CD22 construct of embodiment 11 or embodiment 17 for specific binding to CD22. [0520] 19. The anti-CD22 construct of any one of embodiments 1 to 18, wherein the light chain variable region and the heavy chain variable region are joined by a linker. [0521] 20. The anti-CD22 construct of embodiment 19, wherein the linker has the sequence of SRGGGGSGGGGSGGGGSLEMA (SEQ ID NO: 233). [0522] 21. The anti-CD22 construct of any one of embodiments 1 to 20, wherein the antibody moiety comprises a light chain of the lambda or kappa isotype. [0523] 22. The anti-CD22 construct of any one of embodiments 1 to 21, wherein the antibody moiety binds to a an extracellular region of CD22. [0524] 23. The anti-CD22 construct of embodiment 22, wherein the extracellular region of CD22 comprises at least 7 amino acids of the sequence of DVQYPPKKVTTVIQNPMPIREGDTVTLSCNYNSSNPSVTRYEWKPHGAWEEPSLGVL KIQNVGWDNTTIACAACNSWCSWASPVALNVQYAPRDVRVRKIKPLSEIHSGNSVS LQCDFSSSHPKEVQFFWEKNGRLLGKESQLNFDSISPEDAGSYSCWVNNSIGQTASK AWTLEVLYAPRRLRVSMSPGDQVMEGKSATLTCESDANPPVSHYTWFDWNNQSLP YHSQKLRLEPVKVQHSGAYWCQGTNSVGKGRSPLSTLTVYYSPETIGRR (SEQ ID NO: 205). [0525] 24. The anti-CD22 construct of embodiment 22 or 23, wherein the extracellular region has the sequence of SEQ ID NO: 205. [0526] 25. The anti-CD22 construct of any one of embodiments 1 to 24, wherein the construct is a full-length antibody, a Fab, a Fab, a F(ab)2, an Fv, a single chain Fv (scFv) antibody. [0527] 26. The anti-CD22 construct of any one of embodiments 1 to 25, wherein the construct is monospecific. [0528] 27. The anti-CD22 construct of any one of embodiments 1 to 25, wherein the construct is multispecific. [0529] 28. The anti-CD22 construct of embodiment 27, wherein the construct is bispecific. [0530] 29. The anti-CD22 construct of embodiment 27 or 28, wherein the construct is a tandem scFv, a diabody (Db), a single chain diabody (scDb), a dual-affinity retargeting (DART) antibody, a dual variable domain (DVD) antibody, a knob-into-hole (KiH) antibody, a dock and lock (DNL) antibody, a chemically cross-linked antibody, a heteromultimeric antibody, or a heteroconjugate antibody. [0531] 30. The anti-CD22 construct of embodiment 25, wherein the construct is a tandem scFv comprising two scFvs linked by a peptide linker. [0532] 31. The anti-CD22 construct of embodiment 30, wherein the peptide linker comprises the amino acid sequence of SEQ ID NO: 233. [0533] 32. The anti-CD22 construct of any one of embodiments 27 to 31, wherein the construct further comprises a second antibody moiety that specifically binds to a second antigen. [0534] 33. The anti-CD22 construct of embodiment 32, wherein the second antigen is an antigen on the surface of a T cell. [0535] 34. The anti-CD22 construct of embodiment 33, wherein the T cell is selected from the group consisting of a cytotoxic T cell, a helper T cell, and a natural killer T cell. [0536] 35. The anti-CD22 construct of any one of embodiments 32 to 34, wherein the second antigen is selected from the group consisting of CD3, CD3, CD3, CD3, CD28, CD16a, CD56, CD68, GDS2D, OX40, GITR, CD137, CD27, CD4OL and HVEM. [0537] 36. The anti-CD22 construct of embodiment 35, wherein the second antigen is CD3, and wherein the construct is a tandem scFv comprising an N-terminal scFv specific for CD22 having the sequence of SEQ ID NO: 205 or a portion thereof and a C-terminal scFv specific for CD3. [0538] 37. The anti-CD22 construct of embodiment 32, wherein the second antigen is an antigen on the surface of a natural killer cell, a neutrophil, a monocyte, a macrophage or a dendritic cell. [0539] 38. The anti-CD22 construct of any one of embodiments 1 to 24, wherein the anti-CD22 construct is a chimeric antigen receptor (CAR). [0540] 39. The anti-CD22 construct of embodiment 38, wherein the CAR comprises an anti-CD22 antibody moiety, a transmembrane domain, and an immune cell signaling domain, wherein the anti-CD22 antibody moiety is a scFv comprising the LC-CDR1, the LC-CDR2, and the LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, the HC-CDR1, the HC-CDR2, and the HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively. [0541] 40. The anti-CD22 construct of embodiment 38, wherein the CAR comprises an anti-CD22 antibody moiety, a transmembrane domain, and an immune cell signaling domain, wherein the anti-CD22 antibody moiety is a scFv comprising the LC-CDR1, the LC-CDR2, and the LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, the HC-CDR1, the HC-CDR2, and the HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively. [0542] 41. The anti-CD22 construct of embodiment 39 or 40, wherein the immune cell signaling domain is from a CD3 chain. [0543] 42. The anti-CD22 construct of embodiment 39 or 40, wherein the immune cell signaling domain is from CD28, 4-1BB, ICOS, or OX40. [0544] 43. The anti-CD22 construct of any one of embodiments 39 or 40, wherein the transmembrane domain is a T cell receptor transmembrane domain. [0545] 44. The anti-CD22 construct of any one of embodiments 1 to 24, wherein the anti-CD22 construct is a chimeric antibody-T cell receptor (caTCR) comprising an extracellular domain that binds to CD22 and a T cell receptor (TCR) module (TCRM) comprising TCR transmembrane domains. [0546] 45. The anti-CD22 construct of embodiment 44, wherein the caTCR comprises LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, and HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively. [0547] 46. The anti-CD22 construct of embodiment 44, wherein the caTCR comprises LC-CDR1, LC-CDR2, and LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, and HC-CDR1, HC-CDR2, and HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively. [0548] 47. The anti-CD22 construct of any one of embodiments 44 to 46, wherein the TCRM is capable of recruiting at least one TCR-associated signaling module. [0549] 48. The anti-CD22 construct of embodiment 47, wherein the TCR-associated signaling module is selected from the group consisting of CD3, CD3, and CD3. [0550] 49. The anti-CD22 construct of any one of embodiments 44 to 48, wherein the extracellular domain comprises:

[0551] (a) a first polypeptide comprising a first antigen-binding region comprising a heavy chain variable region (VH) and a CH1 constant domain; and

[0552] (b) a second polypeptide chain comprising a second antigen-binding region comprising a light chain variable region (VL) and a CL constant domain,

[0553] wherein the VH and the CH1 constant domain of the first antigen-binding region and the VL and the CL constant domain of the second antigen-binding region form a Fab-like antigen-binding module that specifically binds to CD22. [0554] 50. The anti-CD22 construct of any one of embodiments 44 to 49, wherein the extracellular domain comprises a scFv that specifically binds to CD22. [0555] 51. The anti-CD22 construct of any one of embodiments 44 to 50, wherein the extracellular domain further comprises at least one additional antibody moiety that specifically binds to at least one non-CD22 antigen. [0556] 52. The anti-CD22 construct of embodiment 51, wherein the at least one non-CD22 antigen is expressed in B-cell malignancy. [0557] 53. The anti-CD22 construct of any one of embodiments 44 to 50, wherein the extracellular domain further comprises an antibody moiety that specifically binds to CD19. [0558] 54. The anti-CD22 construct of any one of embodiments 44 to 50, wherein the extracellular domain further comprises an antibody moiety that specifically binds to CD20. [0559] 55. The anti-CD22 construct of any one of embodiments 44 to 50, wherein the extracellular domain further comprises an antibody moiety that specifically binds to CD19 and an antibody moiety that specifically binds to CD20. [0560] 56. The anti-CD22 construct of any one of embodiments 44 to 55, wherein the caTCR is expressed in combination with a chimeric signaling receptor (CSR). [0561] 57. The anti-CD22 construct of embodiment 56, wherein the CSR comprises an anti-CD22 antibody moiety. [0562] 58. The anti-CD22 construct of embodiment 56, wherein the CSR comprises an antibody moiety that specifically binds a non-CD22 antigen. [0563] 59. The anti-CD22 construct of any one of embodiments 1 to 24, wherein the anti-CD22 construct is a chimeric signaling receptor (CSR). [0564] 60. The anti-CD22 construct of embodiment 59, wherein the CSR comprises

[0565] (a) an anti-CD22 antibody moiety;

[0566] (b) a transmembrane module; and

[0567] (c) a co-stimulatory immune cell signaling module that is capable of providing a co-stimulatory signal to the immune cell,

[0568] wherein the CSR lacks a functional primary immune cell signaling domain. [0569] 61. The anti-CD22 construct of embodiment 60, wherein the anti-CD22 antibody moiety comprises the LC-CDR1, the LC-CDR2, and the LC-CDR3 having the sequences of SEQ ID NOS: 214-216, respectively, the HC-CDR1, the HC-CDR2, and the HC-CDR3 having the sequences of SEQ ID NOS: 209, 210, and 217, respectively. [0570] 62. The anti-CD22 construct of embodiment 60, wherein the anti-CD22 antibody moiety comprises the LC-CDR1, the LC-CDR2, and the LC-CDR3 having the sequences of SEQ ID NOS: 206-208, respectively, the HC-CDR1, the HC-CDR2, and the HC-CDR3 having the sequences of SEQ ID NOS: 209-211, respectively. [0571] 63. The anti-CD22 construct of any one of embodiments 59 to 62, wherein the CSR is expressed in combination with a caTCR or CAR. [0572] 64. The anti-CD22 construct of embodiment 63, wherein the caTCR or CAR specifically targets CD22. [0573] 65. The anti-CD22 construct of embodiment 63, wherein the caTCR or CAR does not specifically target CD22. [0574] 66. The anti-CD22 construct of any one of embodiments 59 to 65, wherein the CSR further comprises at least one additional antibody moiety that specifically binds to at least one non-CD22 antigen. [0575] 67. The anti-CD22 construct of any one of embodiments 56 to 66, wherein the CSR further comprises an antibody moiety that specifically binds to CD19. [0576] 68. The anti-CD22 construct of any one of embodiments 56 to 67, wherein the CSR further comprises an antibody moiety that specifically binds to CD20. [0577] 69. The anti-CD22 construct of any one of embodiments 56 to 68, wherein the CSR comprises a transmembrane fragment and an intracellular fragment that are from the same molecule. [0578] 70. The anti-CD22 construct of embodiment 69, wherein the molecule is selected from the group consisting of CD28, 4-1BB (CD137), OX40, CD30, CD27, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, and a ligand that specifically binds with CD83. [0579] 71. The anti-CD22 construct of embodiment 70, wherein the molecule is selected from the group consisting of CD28, 4-1BB (CD137), OX40, CD30, and CD27. [0580] 72. The anti-CD22 construct of any one of embodiments 56 to 68, wherein the CSR comprises a transmembrane fragment and an intracellular fragment that are from different molecules. [0581] 73. The anti-CD22 construct of embodiment 72, wherein the CSR comprises a transmembrane fragment of a molecule selected from the group consisting of the , , , , or chain of the T-cell receptor, CD28, CD3, CD3, CD45, CD4, CDS, CD8, CD9, CD16, CD22, CD33, CD37, CD64, CD80, CD86, CD134, CD137, and CD154. [0582] 74. The anti-CD22 construct of embodiment 74, wherein the CSR comprises a transmembrane fragment of CD8, 4-1BB, CD27, CD28, CD30, or OX40. [0583] 75. The anti-CD22 construct of any one of embodiments 72 to 75, wherein the transmembrane fragment comprises a sequence of any one of SEQ ID NOS: 145-150. [0584] 76. The anti-CD22 construct of any one of embodiments 72 to 75, wherein the CSR comprises an intracellular fragment of a molecule selected from the group consisting of CD28, 4-1BB (CD137), OX40, CD30, CD27, CD40, PD-1, ICOS, lymphocyte function-associated antigen-1 (LFA-1), CD2, CD7, LIGHT, NKG2C, B7-H3, and a ligand that specifically binds with CD83. [0585] 77. The anti-CD22 construct of embodiment 76, wherein the CSR comprises an intracellular fragment of a molecule selected from the group consisting of CD28, 4-1BB (CD137), OX40, CD30, and CD27. [0586] 78. The anti-CD22 construct of embodiment 76, wherein the intracellular fragment comprises a sequence of any one of SEQ ID NOS: 151-155. [0587] 79. The anti-CD22 construct of any one of embodiments 56 to 78, wherein the CSR comprises a sequence of any one of SEQ ID NOS: 156-171. [0588] 80. The anti-CD22 construct of any one of embodiments 1 to 24, wherein the anti-CD22 construct is an immunoconjugate comprising the antibody moiety and an effector molecule. [0589] 81. The anti-CD22 construct of embodiment 80, wherein the effector molecule is a therapeutic agent selected from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid. [0590] 82. The anti-CD22 construct of embodiment 81, wherein the therapeutic agent is a drug or a toxin. [0591] 83. The anti-CD22 construct of embodiment 80, wherein the effector molecule is a label. [0592] 84. A nucleic acid molecule encoding one or more polypeptides contained in the anti-CD22 construct of any one of embodiments 1 to 83. [0593] 85. The nucleic acid molecule of embodiment 84, wherein the nucleic acid molecule encodes all of the polypeptides contained in the anti-CD22 construct of any one of embodiments 1 to 83. [0594] 86. The nucleic acid molecule of embodiment 85, wherein: [0595] the anti-CD22 construct is a caTCR and is expressed in combination with a CSR, and wherein the nucleic acid molecule encodes all of the polypeptides contained in the caTCR and the polypeptide of the CSR; or [0596] the anti-CD22 construct is a CSR and is expressed in combination with a caTCR or CAR, and wherein the nucleic acid molecule encodes the polypeptide of the CSR and all of the polypeptides contained in the caTCR or CAR. [0597] 87. A set of nucleic acid molecules encoding all of the polypeptides contained in the anti-CD22 construct of any one of embodiments 1 to 83 separately. [0598] 88. The set of nucleic acid molecules of embodiment 87, wherein: [0599] the anti-CD22 construct is a caTCR and is expressed in combination with a CSR, and wherein the set of nucleic acid molecules encode all of the polypeptides contained in the caTCR and the polypeptide of the CSR; or [0600] the anti-CD22 construct is a CSR and is expressed in combination with a caTCR or CAR, and wherein the set of nucleic acid molecules encode the polypeptide of the CSR and all of the polypeptides contained in the caTCR or CAR. [0601] 89. An expression cassette comprising the nucleic acid molecule of any one of embodiments 84 to 86. [0602] 90. A set of expression cassettes comprising nucleic acid molecules encoding all of the polypeptides contained in the anti-CD22 construct of any one of embodiments 1 to 83 separately. [0603] 91. The set of expression cassettes of embodiment 90, wherein the set of expression cassettes comprise the set of nucleic acid molecules of embodiment 87 or 88. [0604] 92. A host cell comprising the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, or the set of expression cassettes of embodiment 90 or 91. [0605] 93. A host cell expressing the anti-CD22 construct of any one of embodiments 1 to 83. [0606] 94. The host cell of embodiment 93, wherein the host cell comprises the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, or the set of expression cassettes of embodiment 90 or 91. [0607] 95. A method of preparing an anti-CD22 construct of any one of embodiments 1 to 83, wherein said method comprising:

[0608] (a) providing a host cell comprising the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, or the set of expression cassettes of embodiment 90 or 91, and

[0609] (b) expressing the nucleic acid molecule(s) or expression cassette(s) in the host cell under conditions that allow for the formation of the anti-CD22 construct. [0610] 96. A pharmaceutical composition comprising a therapeutically effective amount of the anti-CD22 construct of any one of embodiments 1 to 83, the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, the set of expression cassettes of embodiment 90 or 91, or the host cell of any one of embodiments 92 to 94, and one or more pharmaceutically acceptable carriers or excipients. [0611] 97. A method of treating a B-cell malignancy in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the anti-CD22 construct of any one of embodiments 1 to 83, the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, the set of expression cassettes of embodiment 90 or 91, the host cell of any one of embodiments 92 to 94, or the pharmaceutical composition of embodiment 96. [0612] 98. A method of treating a disease or disorder characterized by CD22 overexpression in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the anti-CD22 construct of any one of embodiments 1 to 83, the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, the set of expression cassettes of embodiment 90 or 91, the host cell of any one of embodiments 92 to 94, or the pharmaceutical composition of embodiment 96. In some embodiments of this method, the method is a method of treating a disease. In some embodiments, the method is a method of treating a disorder. In some embodiments, the disease or disorder characterized by CD22 overexpression is cancer (e.g., B-cell malignancy). [0613] 99. A method of treatment comprising introducing the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, or the set of expression cassettes of embodiment 90 or 91 into one or more primary cells isolated from a subject and administering cells comprising the nucleic acid molecule, the set of nucleic acid molecules, the expression cassette, or the set of expression cassettes to the subject. [0614] 100. The method of embodiment 99, further comprising expanding the cells prior to administering the cells to the subject. [0615] 101. The method embodiment 99 or 100, wherein the primary cells are lymphocytes. [0616] 102. The method of embodiment 101, wherein the primary cells are T cells. [0617] 103. A method of detecting CD22 in a sample, comprising: (a) contacting the sample with the anti-CD22 construct of any one of embodiments 1 to 26; and (b) detecting the binding, directly or indirectly, between the anti-CD22 construct and any CD22 in the sample. [0618] 104. The method of embodiment 103, wherein the anti-CD22 construct is conjugated to a detectable label. [0619] 105. The method of embodiment 104, wherein the detectable label is a chromogenic, enzymatic, radioisotopic, isotopic, fluorescent, toxic, chemiluminescent, nuclear magnetic resonance contrast agent. [0620] 106. The method of embodiment 104 or 105, wherein the binding between the anti-CD22 construct and any CD22 in the sample is detected directly by detecting the detectable label. [0621] 107. The method of embodiment 103, wherein the binding between the anti-CD22 construct and any CD22 in the sample is detected indirectly using a secondary antibody. [0622] 108. A method of diagnosing a subject suspected of having a CD22-associated disease or disorder, comprising:

[0623] a) administering an effective amount of the anti-CD22 construct of any one of embodiments 1 to 26 to the subject; and

[0624] b) determining the level of the binding, directly or indirectly, between the anti-CD22 construct and any CD22 in the subject, wherein a level of the binding above a threshold level indicates that the subject has the CD22-associated disease or disorder. [0625] 109. The method of embodiment 108, wherein the CD22-associated disease or disorder is cancer. [0626] 110. The method of embodiment 109, wherein the cancer is a B-cell malignancy. [0627] 111. A method of diagnosing an subject having a B-cell malignancy, comprising: [0628] (a) contacting a sample derived from the subject with the anti-CD22 construct of any one of embodiments 1 to 26 ; and [0629] (b) determining the number of cells bound with the anti-CD22 construct in the sample,

[0630] wherein a value for the number of cells bound with the anti-CD22 construct above a threshold level indicates that the subject has the B-cell malignancy. [0631] 112. The method of embodiment 111, wherein the B-cell malignancy is a CD22+B-cell malignancy. [0632] 113. The method of any one of embodiments 98, 108, and 111, wherein the disease, disorder, or B-cell malignancy is a B-cell lymphoma or a B-cell leukemia. [0633] 114. The method of any one of embodiments 97 to 113, wherein the subject is a human. [0634] 115. Use of the anti-CD22 construct of any one of embodiments 1 to 83, the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, the set of expression cassettes of embodiment 90 or 91, the host cell of any one of embodiments 92 to 94, or the pharmaceutical composition of embodiment 96 for the treatment of a disease or disorder associated with positive CD22 expression. [0635] 116. Use of the anti-CD22 construct of any one of embodiments 1 to 83, the nucleic acid molecule of any one of embodiments 84 to 86, the set of nucleic acid molecules of embodiment 87 or 88, the expression cassette of embodiment 89, the set of expression cassettes of embodiment 90 or 91, the host cell of any one of embodiments 92 to 94, or the pharmaceutical composition of embodiment 96 in the manufacture of a medicament for the treatment of a disease or disorder associated with positive CD22 expression. [0636] 117. Use of the anti-CD22 construct of any one of embodiments 1 to 26 for the diagnosis of a disease or disorder associated with positive CD22 expression. [0637] 118. The use of any one of embodiments 115 to 117, wherein the disease or disorder associated with positive CD22 expression is a cancer.

INFORMAL SEQUENCE LISTING

[0638]

TABLE-US-00009 SEQ ID NO. Sequence Notes 1 METDTLLLWVLLLWVPGSTGEVQLVQSGAEVKKPGESLKISCKGSGYSFTS Construct YWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQ combination1 WSSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSS (anti-CD19-caTCR+ ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHT anti-CD19-CSR_1) FPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEV KTDSTDHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAK TVAVNFLLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETD TLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWY QQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYC QVWDSSSDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLIS DFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKS HRSYSCQVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQL TNTSAYYMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQ AGDVEENPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITC GGNNIGSKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTI SRVEAGDEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGG GSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGL EWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCA RQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSAAAIEVMYPPPYLDN EKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFW VRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 2 METDTLLLWVLLLWVPGSTGQVQLVESGGGLVQPGGSLRLSCAASGFTFSN Construct2 YAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQ (anti-CD22-caTCR) MNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPS SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS SVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKET ENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLF FLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPG STGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYA ASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTR LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVV YFAIITCCLLRRTAFCCNGEKS 3 METDTLLLWVLLLWVPGSTGQVQLVESGGGLVQPGGSLRLSCAASGFTFSN Construct YAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQ combination3 MNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPS (anti-CD22-caTCR+ SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS anti-CD19-CSR_1) SVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKET ENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLF FLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPG STGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYA ASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTR LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVV YFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLL WVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKP GQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVW DSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAE VKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSP SFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQGGMYPRSN WWYNLDSWGQGTLVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPS PLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMT PRRPGPTRKHYQPYAPPRDFAAYRS 4 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct4 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-scFv- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG anti-CD22-caTCR) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKS 5 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination5 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR_1) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLVV YDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVVF GGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKI SCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISA DKSISTAYLQWSSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSW GQGTLVTVSSEQKLISEEDLAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPS PLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMT PRRPGPTRKHYQPYAPPRDFAAYRS 6 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination6 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_1) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVG GVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPP RDFAAYRS 7 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination7 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_2) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDI YIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCS CRFPEEEEGGCEL 8 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination8 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_3) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAATGPADLSPGASSVTPPAPAREPGHSPQIISFFLALTSTALLFLLFFLTLR FSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL 9 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination9 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_4) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDI YIWAPLAGTCGVLLLSLVITLYCQRRKYRSNKGESPVEPAEPCRYSCPREEEG STIPIQEDYRKPEPACSP 10 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination10 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_5) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAATGPTHLPYVSEMLEARTAGHMQTLADFRQLPARTLSTHWPPQRSLC SSDFIRILVIFSGMFLVFTLAGALFLHQRRKYRSNKGESPVEPAEPCRYSCPRE EEGSTIPIQEDYRKPEPACSP 11 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination11 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_6) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDI YIWAPLAGTCGVLLLSLVITLYCHRRACRKRIRQKLHLCYPVQTSQPKLELV DSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDA SPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLA GPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASG K 12 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination12 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_7) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAATGAPPLGTQPDCNPTPENGEAPASTSPTQSLLVDSQASKTLPIPTSAP VALSSTGKPVLDAGPVLFWVILVLVVVVGSSAFLLCHRRACRKRIRQKLHLC YPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSV GAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVG TVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVE EEGKEDPLPTAASGK 13 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination13 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_8) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDI YIWAPLAGTCGVLLLSLVITLYCALYLLRRDQRLPPDAHKPPGGGSFRTPIQE EQADAHSTLAKI 14 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination14 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-scFv- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD22-caTCR+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR_9) QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKSGSGATNFSLLKQAGDVEENPGPMETDTLLLWVLLLWVP GSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIY AASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGT RLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAA SGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSK NTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSEQKLISEE DLAAATGDRDPPATQPQETQGPPARPITVQPTEAWPRTSQGPSTRPVEVPGG RAVAAILGLGLVLGLLGPLAILLALYLLRRDQRLPPDAHKPPGGGSFRTPIQE EQADAHSTLAKI 15 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct15 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-caTCR-1) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY (anti-cd22-scFv+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG GGGGS+anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD19-caTCR) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKS 16 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct16 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-caTCR-2) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY (anti-CD22-scFv+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG 2xGGGGS+anti- QGTLVTVSSGGGGSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSY CD19-caTCR) WIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQW SSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSA STKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTF PAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEV KTDSTDHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAK TVAVNFLLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETD TLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWY QQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYC QVWDSSSDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLIS DFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKS HRSYSCQVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQL TNTSAYYMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKS 17 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct17 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-caTCR-3) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY (anti-CD22-scFv+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG IgCH1+TCRdelta+ QGTLVTVSSTPLGDTTHTSGASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYF anti-CD19-scFv+ PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVN IgCL+ HKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSKSCHKPKAIVHTE TCRgamma)_upper KVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSGSGAPVKQTLNF hingelinker DLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGK TARITCGGNNIGSKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGN TATLTISRVEAGDEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGG GGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQ MPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDT AMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSTPLGDTTHT SGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKA GVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP TECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFA IITCCLLRRTAFCCNGEKS 18 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct18 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-caTCR-4) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY (anti-CD22-scFv+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG IgCH1+TCRdelta+ QGTLVTVSSGGGGSGGGGSGGGGSASTKGPSVFPLAPSSKSTSGGTAALGCL anti-CD19-scFv+ VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT IgCL+ YICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSKSCHKP TCRgamma)_ KAIVHIEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSGSGAPV 3xGGGGSlinker KQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSV SVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFS GSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGG GGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWI GWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSL KASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSGGG GSGGGGSGGGGSQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVA WKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLL LLLKSVVYFAIITCCLLRRTAFCCNGEKS 19 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct19 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-caTCR-5) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY (anti-CD22-scFv+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG IgCH1(S64E, QGTLVTVSSGGGGSGGGGSGGGGSASTKGPSVFPLAPSSKSTSGGTAALGCL S66V)+TCRdelta+ VKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYELVSVVTVPSSSLGTQT anti-CD19-scFv+ YICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSKSCHKP IgCLkappa(S69L, KAIVHIEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSGSGAPV T71S)+ KQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSV TCRgamma)_ SVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFS 3xGGGGSlinker GSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGG GGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWI GWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSL KASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSGGG GSGGGGSGGGGSTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLLSSLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYL LLLLKSVVYFAIITCCLLRRTAFCCNGEKS 20 METDTLLLWVLLLWVPGSTGEVQLVQSGAEVKKPGESLKISCKGSGYSFTS Construct20 YWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQ (anti-CD22-anti- WSSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSS CD19-caTCR-6) GGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQA (anti-CD19VH+ PGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTA anti-CD22VH+ VYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL IgCH1+TCRdelta+ GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG anti-CD19VL+ TQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSKSC anti-CD22VL+ HKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSGSG IgCLkappa+ APVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGLPVLTQP TCRgamma)_ PSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLVVYDDSDRPSGIPE 2xGGGGSlinker RFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVVFGGGTKLTVLGG GGGSGGGGSDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKA PNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLT FGQGTRLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWK VDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLL LLLKSVVYFAIITCCLLRRTAFCCNGEKS 21 METDTLLLWVLLLWVPGSTGQVQLVESGGGLVQPGGSLRLSCAASGFTFSN Construct21 YAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQ (anti-CD22-anti- MNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSGGGGSGGGGSEVQ CD19-caTCR-7) LVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPG (anti-CD22VH+ DSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQ anti-CD19VH+ GGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL IgCH1+TCRdelta+ GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLG anti-CD22VL+ TQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSKSC anti-CD19VL+ HKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSGSG IgCL+ APVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLTQS TCRgamma)_ PSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTGVP 2xGGGGSlinker SRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRGGGG SGGGGSLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVL VVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYV VFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVA WKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHE GSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLL LLLKSVVYFAIITCCLLRRTAFCCNGEKS 22 METDTLLLWVLLLWVPGSTGEVQLVQSGAEVKKPGESLKISCKGSGYSFTS Construct22 YWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQ (anti-CD19-caTCR) WSSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSS ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHT FPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEV KTDSTDHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAK TVAVNFLLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETD TLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWY QQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYC QVWDSSSDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLIS DFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKS HRSYSCQVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQL TNTSAYYMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKS 23 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination23 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR+anti- YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG CD19-CSR) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19scFv+myc DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS tag+truncated VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD28) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (anti-CD22-anti- HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD19-caTCR-1+ LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV anti-CD19-CSR-1A) LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ (construct15+anti- APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS CD19-CSR-1A) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA (construct15+anti- VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC CD19scFv+ QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY truncatedCD28) YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAAIEVMYPPPY LDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFI IFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 24 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct24 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-anti-CD20- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY caTCR-1) YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG (anti-CD22-scFv+ QGTLVTVSSGGGGSLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQK anti-CD19scFv+ PGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVW anti-CD20VH+ DSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAE IgCH1+TCRdelta+ VKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGDSDTRYSP anti-CD20VL+ SFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQGGMYPRSN IgCLkappa+ WWYNLDSWGQGTLVTVSSGGGGSQVQLQQPGAELVKPGASVKMSCKASG TCRgamma+ YTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSS FLAGtag) TAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSSASTKGP SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQQKPGSSP KPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQWTSNPP TFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQW KVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQ GLSSPVTKSFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYL LLLLKSVVYFAIITCCLLRRTAFCCNGEKSDYKDHDGDYKDHDIDYKDDDD K 25 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct25 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-anti-CD20- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY caTCR-2) YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG (anti-CD22scFv+ QGTLVTVSSGGGGSGGGGSQVQLQQPGAELVKPGASVKMSCKASGYTFTS anti-CD20VH+ YNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYM IgCH1+TCRdelta+ QLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSSASTKGPSVFPL anti-CD19scFv+ APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY anti-CD20VL+ SLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKP IgCLkappa+ KETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTA TCRgamma+ KLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLW FLAGtag) VPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLV VYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVV FGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESL KISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGDSDTRYSPSFQGQVTI SADKSISTAYLQWSSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLD SWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASSSVS YIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAA TYYCQQWTSNPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLN NFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEK HKVYACEVTHQGLSSPVTKSFNRGECPIKTDVITMDPKDNCSKDANDTLLLQ LTNTSAYYMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSDYKDHDGDYK DHDIDYKDDDDK 26 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct26 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-anti-CD20- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY caTCR-3) YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG (anti-CD22scFv+ QGTLVTVSSGGGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKMSCKASG anti-CD20VH+ YTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSS IgCH1+TCRdelta+ TAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSSASTKGP anti-CD19scFv+ SVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQ anti-CD20VL+ SSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD IgCLkappa+ HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF TCRgamma+ LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV FLAGtag) LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKK PGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQ GQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQGGMYPRSNWW YNLDSWGQGTLVTVSSGGGGSGGGGSGGGGSQIVLSQSPAILSASPGEKVT MTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLT ISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKS GTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPIKTDVITMDPKDNCS KDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSD YKDHDGDYKDHDIDYKDDDDK 27 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct27 NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (anti-CD22-anti- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-anti-CD20- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY caTCR-4) YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG (anti-CD22scFv+ QGTLVTVSSGGGGSQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHW anti-CD20VH+ VKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLT anti-CD19VH+ SEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSSGGGGSGGGGSEVQLV IgCH1+TCRdelta+ QSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGDS anti-CD20VL+ DTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQGG anti-CD19VL+ MYPRSNWWYNLDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGC IgCL+TCRgamma+ LVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQ FLAGtag) TYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKETENTKQPSKSCHKP KAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSGSGAPV KQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGQIVLSQSPAIL SASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSG SGSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRGGGGSGGG GSLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLVVYD DSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVVFGG GTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKAD GSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTV EKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLK SVVYFAIITCCLLRRTAFCCNGEKSDYKDHDGDYKDHDIDYKDDDDK 28 METDTLLLWVLLLWVPGSTGQVQLQQPGAELVKPGASVKMSCKASGYTFT Construct28 SYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAY (anti-CD20-caTCR) MQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSSASTKGPSVFP (anti-CD20VH+ LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL IgCH1+TCRdelta+ YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVK anti-CD20VL+ PKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLT IgCLkappa+ AKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLL TCRgamma+ WVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPW FLAGtag) IYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGG GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLK SVVYFAIITCCLLRRTAFCCNGEKSDYKDHDGDYKDHDIDYKDDDDK 29 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination29 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-1B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-1B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncatedCD28) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSAAAIEVMYPPPYLDNEKSNGTI IHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSR LLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 30 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination30 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-2A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+ RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-2A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (constmct15+ VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFc+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated4- HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF 1BB) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAATGPADLSPG ASSVTPPAPAREPGHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYI FKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL 31 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination31 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-2B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-2B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncated4-1BB) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSAAATGPADLSPGASSVTPPAPA REPGHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPV QTTQEEDGCSCRFPEEEEGGCEL 32 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination32 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-3A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-3A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated HVKPKETENTKQPSKSCHKPKAIVHtEKVNMMSLTVLGLRMLFAKTVAVNF CD27) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPtECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAATGPTHLPYV SEMLEARTAGHMQTLADFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFL VFTLAGALFLHQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKP EPACSP 33 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination33 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-3B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-3B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncatedCD27) HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSAAATGPTHLPYVSEMLEARTA GHMQTLADFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALF LHQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 34 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination34 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-4A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-4A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD30) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAATGAPPLGTQ PDCNPTPENGEAPASTSPTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGP VLFWVILVLVVVVGSSAFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDS RPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASP AGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGP AEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 35 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination35 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-4B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-4B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncatedCD30) HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSAAATGAPPLGTQPDCNPTPEN GEAPASTSPTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILV LVVVVGSSAFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQ LRSGASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPR DLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEE LEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 36 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination36 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-5A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-5A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF OX40) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAATGDRDPPAT QPQETQGPPARPITVQPTEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLG LLGPLAILLALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 37 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination37 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-5B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-5B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncatedOX40) HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSAAATGDRDPPATQPQETQGPP ARPITVQPTEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLA LYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 38 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination38 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-6A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-6A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+CD8TMand HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD27IC) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAATGTTTPAPR PPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLL LSLVITLYCQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPA CSP 39 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination39 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-6B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-6B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+CD8 SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD TMandCD27IC) HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSAAATGTTTPAPRPPTPAPTIAS QPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYC QRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 40 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination40 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-7A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-7A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+CD8TMand HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD30IC) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAATGTTTPAPR PPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLL LSLVITLYCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGA SVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEP RVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADH TPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 41 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination41 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-7B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-7B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+CD8 SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD TMandCD30IC) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSAAATGTTTPAPRPPTPAPTIAS QPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYC HRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAE ERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTN NKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQE TEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 42 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination42 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-8A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-8A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+CD8TMand HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF OX40IC) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAATGTTTPAPR PPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLL LSLVITLYCALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 43 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination43 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-CSR-8B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD19-CSR-8B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD19scFv+CD8 SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD TMandOX40IC) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSAAATGTTTPAPRPPTPAPTIAS QPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYC ALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 44 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination44 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-1A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-1A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD28) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSEQKLISEEDLAAAIEVMYPPPYLDNEKSNGTIIH VKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRL LHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 45 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination45 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-1B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-1B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncatedCD28) HVKPKEIENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPS PLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMT PRRPGPTRKHYQPYAPPRDFAAYRS 46 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination46 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-2A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-2A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated4- HVKPKEIENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF 1BB) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSEQKLISEEDLAAATGPADLSPGASSVTPPAPAR EPGHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQT TQEEDGCSCRFPEEEEGGCEL 47 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination47 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-2B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-2B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncated4-1BB) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSAAATGPADLSPGASSVTPPAPAREPGHSPQIISF FLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSC RFPEEEEGGCEL 48 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination48 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-3A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-3A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD27) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSEQKLISEEDLAAATGPTHLPYVSEMLEARTAG HMQTLADFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFL HQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 49 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination49 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-3B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-3B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncatedCD27) HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSAAATGPTHLPYVSEMLEARTAGHMQTLADFR QLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLHQRRKYRSN KGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 50 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination50 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-4A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-4A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD30) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSEQKLISEEDLAAATGAPPLGTQPDCNPTPENGE APASTSPTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLV VVVGSSAFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLR SGASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDL PEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELE ADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 51 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination51 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-4B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-4B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncatedCD30) HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSAAATGAPPLGTQPDCNPTPENGEAPASTSPTQS LLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSSAFLL CHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVA EERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHT NNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQ ETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 52 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination52 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-5A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD229-CSR-5A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+truncated HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF OX40) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSEQKLISEEDLAAATGDRDPPATQPQETQGPPAR PITVQPTEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALY LLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 53 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination53 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-5B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-5B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+ SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD truncatedOX40) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSAAATGDRDPPATQPQETQGPPARPITVQPILA WPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALYLLRRDQRL PPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 54 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination54 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-6A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-6A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+CD8TMand HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD27IC) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQP LSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQR RKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 55 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination55 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-6B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-6B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+CD8 SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD TMandCD27IC) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRP AAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSNKGES PVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 56 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination56 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-7A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-7A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+CD8TMand HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD30IC) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQP LSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHR RACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEER GLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNK IEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETE PPLGSCSDVMLSVEEEGKEDPLPTAASGK 57 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination57 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-7B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-7B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+CD8 SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD TMandCD30IC) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRP AAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRACRKRIRQK LHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMET CHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADT VIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDVM LSVEEEGKEDPLPTAASGK 58 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination58 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-caTCR-1 GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY (anti-CD22-anti- YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-8A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-8A) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+myc SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD tag+CD8TMand HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF OX40IC) LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQP LSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCAL YLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 59 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination59 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD22-CSR-8B) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (construct15+anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS CD22-CSR-8B) DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS (construct15+anti- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CD22scFv+CD8 SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD TMandOX40IC) HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDI RNYLNWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPE DIATYYCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQ LVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGS GGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAW MDSWGQGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRP AAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRDQRLP PDAHKPPGGGSFRTPIQEEQADAHSTLAKI 60 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination60 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-1A) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-1A) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+myc LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ tag+truncated APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS CD28) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAAIEVMYPPP YLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVA FIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 61 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination61 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-1B) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-1B) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+ LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ truncatedCD28) APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSAAAIEVMYPPPYLDNEKSNG TIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFBFWVRSKR SRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 62 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination62 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-2A) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-2A) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+myc LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ tag+truncated4- APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS 1BB) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGPADLS PGASSVTPPAPAREPGHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLL YIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL 63 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination63 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-2B) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-2B) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+ LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ truncated4-1BB) APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSAAATGPADLSPGASSVTPPA PAREPGHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRP VQTTQEEDGCSCRFPEEEEGGCEL 64 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination64 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-3A) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-3A) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+myc LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ tag+truncated APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS CD27) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGPTHLP YVSEMLEARTAGHMQTLADFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGM FLVFTLAGALFLHQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYR KPEPACSP 65 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination65 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-3B) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-3B) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+ LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ truncatedCD27) APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSAAATGPTHLPYVSEMLEART AGHMQTLADFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGAL FLHQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 66 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination66 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-4A) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-4A) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+myc LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ tag+truncated APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS CD30) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGAPPLG TQPDCNPTPENGEAPASTSPTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDA GPVLFWVILVLVVVVGSSAFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELV DSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDA SPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLA GPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASG K 67 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination67 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-4B) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-4B) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+ LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ truncatedCD30) APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSAAATGAPPLGTQPDCNPTPE NGEAPASTSPTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVIL VLVVVVGSSAFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSST QLRSGASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSP RDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEE ELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 68 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination68 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-5A) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-5A) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKEIENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+myc LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ tag+truncated APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS OX40) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGDRDPP ATQPQETQGPPARPITVQPTEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLV LGLLGPLAILLALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 69 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination69 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-5B) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-5B) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKEIENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+ LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ truncatedOX40) APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSAAATGDRDPPATQPQETQGP PARPITVQPTEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILL ALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 70 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination70 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-6A) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-6A) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+myc LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ tag+CD8TMand APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS CD27IC) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGTTTPAP RPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVL LLSLVITLYCQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEP ACSP 71 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination71 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-6B) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-6B) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+CD8 LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ TMandCD27IC) APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSAAATGTTTPAPRPPTPAPTIA SQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYC QRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 72 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination72 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-7A) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-7A) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKEIENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+myc LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ tag+CD8TMand APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS CD30IC) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGTTTPAP RPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVL LLSLVITLYCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSG ASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPE PRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEAD HTPHYPEQEIEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 73 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination73 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-7B) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-7B) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+CD8 LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ TMandCD30IC) APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSAAATGTTTPAPRPPTPAPTIA SQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYC HRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAE ERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTN NKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQE TEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 74 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination74 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD19-caTCR-1+ GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY anti-CD19-anti- YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG CD20-CSR-8A) QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV (anti-CD22-anti- RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-8A) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWV CD20scFv+myc LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ tag+CD8TMand APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS OX40IC) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDS AVYYCARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGTTTPAP RPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVL LLSLVITLYCALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 75 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Construct NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY combination75 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG (anti-CD22-anti- GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY CD19-caTCR-1+ YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG anti-CD19-anti- QGTLVTVSSGGGGSEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWV CD20-CSR-8B) RQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKAS (construct15+anti- DTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSSASTKGPS CD19-anti-CD20- VFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQS CSR-8B) SGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTD (construct15+anti- HVKPKEIENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLFAKTVAVNF CD19scFv+anti- LLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQ CD20scFv+CD8 APVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSS TMandOX40IC) SDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGA VTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSC QVTHEGSTVEKTVAPTECSPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAY YMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKSGSGATNFSLLKQAGDVEE NPGPMETDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIG SKSVHWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAG DEADYYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEM AEVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGI IYPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG WQGGMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILS ASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGS GSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGG GGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQ TPGRGLEW IGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARS TYYGGDWYFNVWGAGTTVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPE ACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRD QRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 76 METDTLLLWVLLLWVPGSTGEVQLVQSGAEVKKPGESLKISCKGSGYSFTS Anti-CD19-caTCR YWIGWVRQMPGKGLEWMGIIYPGDSDTRYSPSFQGQVTISADKSISTAYLQ (SP-anti-CD19VH- WSSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQGTLVTVSS IgCH1-TCRdelta- ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHT F2Apeptide-SP- FPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEV anti-CD19VL- KTDSTDHVKPKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAK IgCL-TCRgamma) TVAVNFLLTAKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETD TLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWY QQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYC QVWDSSSDYVVFGGGTKLTVLGQPKANPTVTLFPPSSEELQANKATLVCLIS DFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNKYAASSYLSLTPEQWKS HRSYSCQVTHEGSTVEKTVAP1ECSPIKTDVITMDPKDNCSKDANDTLLLQL TNTSAYYMYLLLLLKSVVYFAIITCCLLRRTAFCCNGEKS 77 METDTLLLWVLLLWVPGSTGQVQLVESGGGLVQPGGSLRLSCAASGFTFSN Anti-CD22- YAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQ caTCR_monovalent MNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPS (SP-anti-CD22VH- SKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLS IgCH1-TCRdelta- SVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKET F2Apeptide-SP- ENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLF anti-CD22VL- FLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPG IgCLkappa- STGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYA TCRgamma) ASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTR LEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTK SFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVV YFAIITCCLLRRTAFCCNGEKS 78 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22- NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY caTCR_bivalent YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWV RQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAE DTAVYYCARYGSAAWMDSWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGT AALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSS SLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVKPKEIENTKQPSK SCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLTAKLFFLRAKRSG SGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLLWVPGSTGDIQLT QSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASNLQTG VPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESV TEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPI KTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITCCL LRRTAFCCNGEKS 79 METDTLLLWVLLLWVPGSTGQVQLQQPGAELVKPGASVKMSCKASGYTFT Anti-CD20-caTCR SYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAY (SP-anti-CD20VH- MQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSSASTKGPSVFP IgCH1-TCRdelta- LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL F2Apeptide-SP- YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVK anti-CD20VL- PKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLT IgCLkappa- AKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLL TCRgamma)_no WVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPW FLAGtag IYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGG GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLK SVVYFAIITCCLLRRTAFCCNGEKS 80 METDTLLLWVLLLWVPGSTGQVQLQQPGAELVKPGASVKMSCKASGYTFT Anti-CD20-caTCR SYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAY (SP-anti-CD20VH- MQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSSASTKGPSVFP IgCH1-TCRdelta- LAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGL F2Apeptide-SP- YSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCEVKTDSTDHVK anti-CD20VL- PKETENTKQPSKSCHKPKAIVHTEKVNMMSLTVLGLRMLFAKTVAVNFLLT IgCLkappa- AKLFFLRAKRSGSGAPVKQTLNFDLLKLAGDVESNPGPMETDTLLLWVLLL TCRgamma-FLAG WVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPW tag) IYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGG GTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDN ALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSP VTKSFNRGECPIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLK SVVYFAIITCCLLRRTAFCCNGEKSDYKDHDGDYKDHDIDYKDDDDK 81 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-1A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncatedCD28) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGPADLSPGASSVTPPAPAREPGHSP QIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEED GCSCRFPEEEEGGCEL 82 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-1B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD28) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGPADLSPGASSVTPPAPAREPGHSPQIISFFLALTS TALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEE EGGCEL 83 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-2A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncated4-1BB) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGPTHLPYVSEMLEARTAGHMQTLA DFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLHQRRKY RSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 84 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-2B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (truncated4-1BB) YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGPTHLPYVSEMLEARTAGHMQTLADFRQLPART LSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLHQRRKYRSNKGESPVE PAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 85 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-3A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncatedCD27) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGAPPLGTQPDCNPTPENGEAPASTS PTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSS AFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVT EPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVS TEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPH YPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 86 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-3B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD27) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGAPPLGTQPDCNPTPENGEAPASTSPTQSLLVDSQ ASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSSAFLLCHRRAC RKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASV1EPVAEERGLM SQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKI YIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLG SCSDVMLSVEEEGKEDPLPTAASGK 87 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-4A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncatedCD30) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGDRDPPATQPQETQGPPARPITVQP TEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALYLLRRD QRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 88 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-4B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD30) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGDRDPPATQPQETQGPPARPITVQPTEAWPRTSQ GPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALYLLRRDQRLPPDAHK PPGGGSFRTPIQEEQADAHSTLAKI 89 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-5A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncatedOX40) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPE ACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSN KGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 90 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-5B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG OX40) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGA VHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSNKGESPVEPA EPCRYSCPREEEGSTIPIQEDYRKPEPACSP 91 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-6A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+CD8 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG TMandCD27IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPE ACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRACRK RIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQP LMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIM KADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCS DVMLSVEEEGKEDPLPTAASGK 92 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-6B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withCD8TMand YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD27IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGA VHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRACRKRIRQKLHLCY PVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVG AAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGT VKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEE EGKEDPLPTAASGK 93 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-7A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+CD8 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG TMandCD30IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPE ACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRD QRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 94 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD19-CSR-7B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withCD8TMand YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD30IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGA VHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRDQRLPPDAHK PPGGGSFRTPIQEEQADAHSTLAKI 95 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-CSR-8A HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD (withmyctag+CD8 YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL TMandOX40IC) VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSEQKLISEEDLAAATGTTTPAPRPPTP APTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLV ITLYCALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 96 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-CSR-8B HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD (withCD8TMand YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL OX40IC) VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSAAATGTTTPAPRPPTPAPTIASQPLS LRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYL LRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 97 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-1A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncatedCD28) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAAIEVMYPPPYLDNEKSNGTIIHVKGKH LCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDY MNMTPRRPGPTRKHYQPYAPPRDFAAYRS 98 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-1B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD28) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGP SKPFWVLVVVGGVLACYSLLVTVAFBFWVRSKRSRLLHSDYMNMTPRRPG PTRKHYQPYAPPRDFAAYRS 99 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-2A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncated4-1BB) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGPADLSPGASSVTPPAPAREPGHSP QIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEED GCSCRFPEEEEGGCEL 100 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-2B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated4- YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG 1BB) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGPADLSPGASSVTPPAPAREPGHSPQIISFFLALTS TALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEE EGGCEL 101 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-3A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncatedCD27) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGPTHLPYVSEMLEARTAGHMQTLA DFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLHQRRKY RSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 102 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-3B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD27) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGPTHLPYVSEMLEARTAGHMQTLADFRQLPART LSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLHQRRKYRSNKGESPVE PAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 103 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-4A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncatedCD30) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGAPPLGTQPDCNPTPENGEAPASTS PTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSS AFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVT EPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVS TEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPH YPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 104 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-4B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD30) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGAPPLGTQPDCNPTPENGEAPASTSPTQSLLVDSQ ASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSSAFLLCHRRAC RKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLM SQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKI YIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLG SCSDVMLSVEEEGKEDPLPTAASGK 105 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-5A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+ YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG truncatedOX40) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGDRDPPATQPQETQGPPARPITVQP TEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALYLLRRD QRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 106 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-5B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withtruncated YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG OX40) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGDRDPPATQPQETQGPPARPITVQPTEAWPRTSQ GPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALYLLRRDQRLPPDAHK PPGGGSFRTPIQEEQADAHSTLAKI 107 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-6A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+CD8 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG TMandCD27IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPE ACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSN KGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 108 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-6B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withCD8TMand YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD27IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGA VHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSNKGESPVEPA EPCRYSCPREEEGSTIPIQEDYRKPEPACSP 109 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-7A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+CD8 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG TMandCD30IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPE ACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRACRK RIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQP LMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIM KADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCS DVMLSVEEEGKEDPLPTAASGK 110 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-7B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withCD8TMand YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG CD30IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGA VHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRACRKRIRQKLHLCY PVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVG AAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGT VKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEE EGKEDPLPTAASGK 111 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-8A NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withmyctag+CD8 YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG TMandOX40IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPE ACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRD QRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 112 METDTLLLWVLLLWVPGSTGDIQLTQSPSSLSTSVGDRVTITCQASHDIRNYL Anti-CD22-CSR-8B NWYQQKPGKAPNLLIYAASNLQTGVPSRFSGRGSGTDFTLTISSLQPEDIATY (withCD8TMand YCQQYDGLPLTFGQGTRLEIKRSRGGGGSGGGGSGGGGSLEMAQVQLVESG OX40IC) GGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSISGSGGSTY YADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSAAWMDSWG QGTLVTVSSVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGA VHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRDQRLPPDAHK PPGGGSFRTPIQEEQADAHSTLAKI 113 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-1A WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withmyctag+ YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE truncatedCD28) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSEQKLISEEDLAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLC PSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMN MTPRRPGPTRKHYQPYAPPRDFAAYRS 114 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-1B WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withtruncated YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE CD28) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKP FWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTR KHYQPYAPPRDFAAYRS 115 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-2A WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withmyctag+ YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE truncated4-1BB) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSEQKLISEEDLAAATGPADLSPGASSVTPPAPAREPGHSPQIIS FFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCS CRFPEEEEGGCEL 116 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-2B WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withtruncated4- YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE 1BB) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSAAATGPADLSPGASSVTPPAPAREPGHSPQIISFFLALTSTA LLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEG GCEL 117 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-3A WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (wimmyctag+ YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE truncatedCD27) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSEQKLISEEDLAAATGPTHLPYVSEMLEARTAGHMQTLADF RQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLHQRRKYRS NKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 118 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-3B WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withtruncated YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE CD27) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSAAATGPTHLPYVSEMLEARTAGHMQTLADFRQLPARTLST HWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLHQRRKYRSNKGESPVEPA EPCRYSCPREEEGSTIPIQEDYRKPEPACSP 119 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-4A WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withmyctag+ YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE truncatedCD30) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSEQKLISEEDLAAATGAPPLGTQPDCNPTPENGEAPASTSPT QSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSSAF LLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEP VAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTE HTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYP EQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 120 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-4B WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withtruncated YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE CD30) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSAAATGAPPLGTQPDCNPTPENGEAPASTSPTQSLLVDSQAS KTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSSAFLLCHRRACR KRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMS QPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIY IMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGS CSDVMLSVEEEGKEDPLPTAASGK 121 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-5A WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withmyctag+ YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE truncatedOX40) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSEQKLISEEDLAAATGDRDPPATQPQETQGPPARPITVQPTE AWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALYLLRRDQR LPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 122 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-5B WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withtruncated YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE OX40) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSAAATGDRDPPATQPQETQGPPARPITVQPTEAWPRTSQGPS TRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALYLLRRDQRLPPDAHKPPG GGSFRTPIQEEQADAHSTLAKI 123 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-6A WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withmyctag+CD8 YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE TMandCD27IC) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACR PAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSNKGE SPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 124 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-6B WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withCD8TMand YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE CD27IC) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVH TRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSNKGESPVEPAEPC RYSCPREEEGSTIPIQEDYRKPEPACSP 125 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-7A WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withmyctag+CD8 YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE TMandCD30IC) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACR PAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRACRKRIRQ KLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLME TCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKAD TVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDV MLSVEEEGKEDPLPTAASGK 126 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-7B WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withCD8TMand YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE CD30IC) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVH TRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRACRKRIRQKLHLCYPV QTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVGAA YLESLPLQDASPAGGPSSPRDLPEPRVS1EHTNNKIEKIYIMKADTVIVGTVK AELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEEEG KEDPLPTAASGK 127 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-8A WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withmyctag+CD8 YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE TMandOX40IC) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSEQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACR PAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRDQRL PPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 128 METDTLLLWVLLLWVPGSTGQIVLSQSPAILSASPGEKVTMTCRASSSVSYIH Anti-CD20-CSR-8B WFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATY (withCD8TMand YCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAE OX40IC) LVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSY NQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNV WGAGTTVTVSSAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVH TRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRDQRLPPDAHKPP GGGSFRTPIQEEQADAHSTLAKI 129 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-1A YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withmyctag+ VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD truncatedCD28) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAAIEVMYPPPYLDN EKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFW VRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 130 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-1B YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withtruncated VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD CD28) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSAAAIEVMYPPPYLDNEKSNGTIIHV KGKHLCPSPLFPGPSKPFWVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLL HSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS 131 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-2A YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withmyctag+ VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD truncated4-1BB) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGPADLSPGASS VTPPAPAREPGHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQ PFMRPVQTTQEEDGCSCRFPEEEEGGCEL 132 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-2B YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withtruncated4- VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD 1BB) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSAAATGPADLSPGASSVTPPAPAREP GHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTT QEEDGCSCRFPEEEEGGCEL 133 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-3A YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withmyctag+ VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD truncatedCD27) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGPTHLPYVSEM LEARTAGHMQTLADFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFT LAGALFLHQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPA CSP 134 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-3B YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withtruncated VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD CD27) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSAAATGPTHLPYVSEMLEARTAGH MQTLADFRQLPARTLSTHWPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLH QRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 135 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-4A YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withmyctag+ VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD truncatedCD30) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGAPPLGTQPDC NPTPENGEAPASTSPTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLF WVILVLVVVVGSSAFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPR RSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGG PSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEP ELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 136 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-4B YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withtruncated VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD CD30) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSAAATGAPPLGTQPDCNPTPENGEA PASTSPTQSLLVDSQASKTLPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVV VVGSSAFLLCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRS GASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLP EPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEA DHTPHYPEQE1EPPLGSCSDVMLSVEEEGKEDPLPTAASGK 137 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-5A YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withmyctag+ VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD truncatedOX40) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGDRDPPATQPQ ETQGPPARPITVQPTEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGP LAILLALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 138 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-5B YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withtruncated VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD OX40) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSAAATGDRDPPATQPQETQGPPARPI TVQPTEAWPRTSQGPSTRPVEVPGGRAVAAILGLGLVLGLLGPLAILLALYL LRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 139 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-6A YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withmyctag+CD8 VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD TMandCD27IC) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGTTTPAPRPPT PAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSL VITLYCQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 140 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-6B YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withCD8TMand VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD CD27IC) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSAAATGTTTPAPRPPTPAPTIASQPLS LRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRK YRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP 141 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-7A YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withmyctag+CD8 VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD TMandCD30IC) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGTTTPAPRPPT PAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSL VITLYCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVT EPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVS TEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPH YPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 142 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-7B YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withCD8TMand VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD CD30IC) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSAAATGTTTPAPRPPTPAPTIASQPLS LRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRA CRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGL MSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIE KIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQE1EPP LGSCSDVMLSVEEEGKEDPLPTAASGK 143 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-8A YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withmyctag+CD8 VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD TMandOX40IC) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSEQKLISEEDLAAATGTTTPAPRPPT PAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSL VITLYCALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 144 METDTLLLWVLLLWVPGSTGLPVLTQPPSVSVAPGKTARITCGGNNIGSKSV Anti-CD19-anti- HWYQQKPGQAPVLVVYDDSDRPSGIPERFSGSNSGNTATLTISRVEAGDEAD CD20-CSR-8B YYCQVWDSSSDYVVFGGGTKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQL (withCD8TMand VQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGITYPGD OX40IC) SDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWGWQG GMYPRSNWWYNLDSWGQGTLVTVSSGGGGSGGGGSQIVLSQSPAILSASPG EKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNLASGVPVRFSGSGSGT SYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIKRSRGGGGSGGGGSG GGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRG LEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYY CARSTYYGGDWYFNVWGAGTTVTVSSAAATGTTTPAPRPPTPAPTIASQPLS LRPEACRPAAGGAVHTRGLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYL LRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI 145 IYIWAPLAGTCGVLLLSLVIT CD8transmembrane (TM)sequence 146 IISFFLALTSTALLFLLFFLTLRFSVV 4-1BBTMsequence 147 ILVIFSGMFLVFTLAGALFLH CD27TMsequence 148 FWVLVVVGGVLACYSLLVTVAFIIFWV CD28TMsequence 149 PVLDAGPVLFWVILVLVVVVGSSAFLLC CD30TMsequence 150 VAAILGLGLVLGLLGPLAILL OX40TMsequence 151 KRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL 4-1BBICsignaling sequence 152 QRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQEDYRKPEPACSP CD27ICsignaling sequence 153 RSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAAYRS CD28ICsignaling sequence 154 HRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAE CD30ICsignaling ERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTN sequence NKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQE TEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 155 ALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHSTLAKI OX40ICsignaling sequence 156 EQKLISEEDLAAAIEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPF myctag+truncated WVLVVVGGVLACYSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRK CD28-1A HYQPYAPPRDFAAYRS 157 IEVMYPPPYLDNEKSNGTIIHVKGKHLCPSPLFPGPSKPFWVLVVVGGVLAC truncatedCD28-1B YSLLVTVAFIIFWVRSKRSRLLHSDYMNMTPRRPGPTRKHYQPYAPPRDFAA YRS 158 EQKLISEEDLAAATGPADLSPGASSVTPPAPAREPGHSPQIISFFLALTSTALLF myctag+truncated LLFFLTLRFSVVKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCE 4-1BB-2A L 159 PADLSPGASSVTPPAPAREPGHSPQIISFFLALTSTALLFLLFFLTLRFSVVKRG truncated4-1BB-2B RKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEEGGCEL 160 EQKLISEEDLAAATGPTHLPYVSEMLEARTAGHMQTLADFRQLPARTLSTH myctag+truncated WPPQRSLCSSDFIRILVIFSGMFLVFTLAGALFLHQRRKYRSNKGESPVEPAEP CD27-3A CRYSCPREEEGSTIPIQEDYRKPEPACSP 161 PTHLPYVSEMLEARTAGHMQTLADFRQLPARTLSTHWPPQRSLCSSDFIRILV truncatedCD27-3B IFSGMFLVFTLAGALFLHQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQ EDYRKPEPACSP 162 EQKLISEEDLAAATGAPPLGTQPDCNPTPENGEAPASTSPTQSLLVDSQASKT myctag+truncated LPIPTSAPVALSSTGKPVLDAGPVLFWVILVLVVVVGSSAFLLCHRRACRKRI CD30-4A RQKLHLCYPVQTSQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPL METCHSVGAAYLESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMK ADTVIVGTVKAELPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSD VMLSVEEEGKEDPLPTAASGK 163 APPLGTQPDCNPTPENGEAPASTSPTQSLLVDSQASKTLPIPTSAPVALSSTGK truncatedCD30-4B PVLDAGPVLFWVILVLVVVVGSSAFLLCHRRACRKRIRQKLHLCYPVQTSQP KLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVGAAYLESL PLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPE GRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLP TAASGK 164 EQKLISEEDLAAATGDRDPPATQPQETQGPPARPITVQPTEAWPRTSQGPSTR myctag+truncated PVEVPGGRAVAAILGLGLVLGLLGPLAILLALYLLRRDQRLPPDAHKPPGGG OX40-5A SFRTPIQEEQADAHSTLAKI 165 DRDPPATQPQETQGPPARPITVQP1EAWPRTSQGPSTRPVEVPGGRAVAAILG truncatedOX40-5B LGLVLGLLGPLAILLALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHST LAKI 166 EQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTR myctag+CD8TM GLDFACDIYIWAPLAGTCGVLLLSLVITLYCQRRKYRSNKGESPVEPAEPCR ICandCD27-6A YSCPREEEGSTIPIQEDYRKPEPACSP 167 TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLA CD8TMandCD27 GTCGVLLLSLVITLYCQRRKYRSNKGESPVEPAEPCRYSCPREEEGSTIPIQED IC6B YRKPEPACSP 168 EQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTR myctag+CD8TM GLDFACDIYIWAPLAGTCGVLLLSLVITLYCHRRACRKRIRQKLHLCYPVQT andCD30IC-7A SQPKLELVDSRPRRSSTQLRSGASVTEPVAEERGLMSQPLMETCHSVGAAYL ESLPLQDASPAGGPSSPRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAE LPEGRGLAGPAEPELEEELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKE DPLPTAASGK 169 TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLA CD8TMandCD30 GTCGVLLLSLVITLYCHRRACRKRIRQKLHLCYPVQTSQPKLELVDSRPRRSS 1C-7B TQLRSGASVTEPVAEERGLMSQPLMETCHSVGAAYLESLPLQDASPAGGPSS PRDLPEPRVSTEHTNNKIEKIYIMKADTVIVGTVKAELPEGRGLAGPAEPELE EELEADHTPHYPEQETEPPLGSCSDVMLSVEEEGKEDPLPTAASGK 170 EQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTR myctag+CD8TM GLDFACDIYIWAPLAGTCGVLLLSLVITLYCALYLLRRDQRLPPDAHKPPGG ICandOX40-8A GSFRTPIQEEQADAHSTLAKI 171 TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLA CD8TMandOX40 GTCGVLLLSLVITLYCALYLLRRDQRLPPDAHKPPGGGSFRTPIQEEQADAHS IC-8B TLAKI 172 EQKLISEEDLAAATGTTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTR myctag+CD8TM GLDFACDIYIWAPLAGTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTT sequenceand4-1BB QEEDGCSCRFPEEEEGGCEL ICsignaling sequence 173 TTTPAPRPPTPAPTIASQPLSLRPEACRPAAGGAVHTRGLDFACDIYIWAPLA CD8TMsequence GTCGVLLLSLVITLYCKRGRKKLLYIFKQPFMRPVQTTQEEDGCSCRFPEEEE and4-1BBIC GGCEL signalingsequence 174 EVQLVQSGAEVKKPGESLKISCKGSGYSFTSYWIGWVRQMPGKGLEWMGII YPGDSDTRYSPSFQGQVTISADKSISTAYLQWSSLKASDTAMYYCARQVWG Anti-CD19VH WQGGMYPRSNWWYNLDSWGQGTLVTVSS 175 LPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLVVYDDS Anti-CD19VL DRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVVFGGGT KLTVL 176 QVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSI Anti-CD22VH SGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSA AWMDSWGQGTLVTVSS 177 DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASN Anti-CD22VL LQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIK R 178 QVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGA Anti-CD20VH IYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTY YGGDWYFNVWGAGTTVTVSS 179 QIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNL Anti-CD20VL ASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIK R 180 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHT IgCH1 FPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC 181 ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHT IgCH1(S64E, FPAVLQSSGLYELVSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSC S66V) GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKA 182 GVETTKPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAP IgCL TECS 183 TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS IgCLkappa QESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR GEC 184 TVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNS IgCLkappa(S69L, QESVTEQDSKDSTYSLLSSLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNR T71S) GEC 185 LPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLVVYDDS Anti-CD19scFv DRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVVFGGGT (anti-CD19VL+ KLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKG linker+anti-CD19 SGYSFTSYWIGWVRQMPGKGLEWMGITYPGDSDTRYSPSFQGQVTISADKSI VH) STAYLQWSSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQG TLVTVSS 186 DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASN Anti-CD22scFv LQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIK (anti-CD22VL+ RSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAASGFTF linkeranti-CD22 SNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLY VH) LQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSS 187 QIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQQKPGSSPKPWIYATSNL Anti-CD20say ASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQWTSNPPTFGGGTKLEIK (anti-CD20VL+ RSRGGGGSGGGGSGGGGSLEQVQLQQPGAELVKPGASVKMSCKASGYTFT linker+anti-CD20 SYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAY VH) MQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAGTTVTVSS 188 LPVLTQPPSVSVAPGKTARITCGGNNIGSKSVHWYQQKPGQAPVLVVYDDS Anti-CD19scFv- DRPSGIPERFSGSNSGNTATLTISRVEAGDEADYYCQVWDSSSDYVVFGGGT linker-anti-CD20 KLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVQSGAEVKKPGESLKISCKG scFv SGYSFTSYWIGWVRQMPGKGLEWMGITYPGDSDTRYSPSFQGQVTISADKSI STAYLQWSSLKASDTAMYYCARQVWGWQGGMYPRSNWWYNLDSWGQG TLVTVSSGGGGSGGGGSQIVLSQSPAILSASPGEKVTMTCRASSSVSYIHWFQ QKPGSSPKPWIYATSNLASGVPVRFSGSGSGTSYSLTISRVEAEDAATYYCQQ WTSNPPTFGGGTKLEIKRSRGGGGSGGGGSGGGGSLEQVQLQQPGAELVKP GASVKMSCKASGYTFTSYNMHWVKQTPGRGLEWIGAIYPGNGDTSYNQKF KGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARSTYYGGDWYFNVWGAG TTVTVSS 189 METDTLLLWVLLLWVPGSTG Signalpeptide(SP) 190 RAKRSGSGAPVKQTLNFDLLKLAGDVESNPGP F2Apeptide 191 GSGATNFSLLKQAGDVEENPGP P2A 192 EVKTDSTDHVKPKETENTKQPSKSCHKPKAIVHIEKVNMMSLTVLGLRMLF TCRdelta AKTVAVNFLLTAKLFFL 193 PIKTDVITMDPKDNCSKDANDTLLLQLTNTSAYYMYLLLLLKSVVYFAIITC CLLRRTAFCCNGEKS TCRgamma 194 EQKLISEEDL Myctag 195 DYKDHDGDYKDHDIDYKDDDDK FLAGtag 196 GSRGGGGSGGGGSGGGGSLEMA Linker 197 SRGGGGSGGGGSGGGGSLEMA Linker 198 GGGGS Linker 199 GGGGSGGGGS Linker 200 GGGGSGGGGSGGGGS Linker 201 AAATG Linker 202 AAA Linker 203 TPLGDTTHTSG Linker(IgG3upper hinge 204 MHLLGPWLLLLVLEYLAFSDSSKWVFEHPETLYAWEGACVWIPCTYRALD full-lengthhuman GDLESFILFHNPEYNKNTSKFDGTRLYESTKDGKVPSEQKRVQFLGDKNKNC CD22 TLSIHPVHLNDSGQLGLRMESKTEKWMERIHLNVSERPFPPHIQLPPEIQESQE VTLTCLLNFSCYGYPIQLQWLLEGVPMRQAAVTSTSLTIKSVFTRSELKFSPQ WSHHGKIVTCQLQDADGKFLSNDTVQLNVKHTPKLEIKVTPSDAIVREGDS VTMTCEVSSSNPEYTTVSWLKDGTSLKKQNTFTLNLREVTKDQSGKYCCQV SNDVGPGRSEEVFLQVQYAPEPSTVQILHSPAVEGSQVEFLCMSLANPLPTN YTWYHNGKEMQGRTEEKVHIPKILPWHAGTYSCVAENILGTGQRGPGAELD VQYPPKKVTTVIQNPMPIREGDTVTLSCNYNSSNPSVTRYEWKPHGAWEEPS LGVLKIQNVGWDNTTIACAACNSWCSWASPVALNVQYAPRDVRVRKIKPLS EIHSGNSVSLQCDFSSSHPKEVQFFWEKNGRLLGKESQLNFDSISPEDAGSYS CWVNNSIGQTASKAWTLEVLYAPRRLRVSMSPGDQVMEGKSATLTCESDA NPPVSHYTWFDWNNQSLPYHSQKLRLEPVKVQHSGAYWCQGTNSVGKGRS PLSTLTVYYSPETIGRRVAVGLGSCLAILILAICGLKLQRRWKRTQSQQGLQE NSSGQSFFVRNKKVRRAPLSEGPHSLGCYNPMMEDGISYTTLRFPEMNIPRT GDAESSEMQRPPPDCDDTVTYSALHKRQVGDYENVIPDFPEDEGIHYSELIQF GVGERPQAQENVDYVILKH 205 DVQYPPKKVTTVIQNPMPIREGDTVTLSCNYNSSNPSVTRYEWKPHGAWEE extracellularregion PSLGVLKIQNVGWDNTTIACAACNSWCSWASPVALNVQYAPRDVRVRKIKP containingdomains LSEIHSGNSVSLQCDFSSSHPKEVQFFWEKNGRLLGKESQLNFDSISPEDAGS 5-7ofCD22 YSCWVNNSIGQTASKAWTLEVLYAPRRLRVSMSPGDQVMEGKSATLTCES DANPPVSHYTWFDWNNQSLPYHSQKLRLEPVKVQHSGAYWCQGTNSVGK GRSPLSTLTVYYSPETIGRR 206 SSNIGNNY Clone1LC-CDR1 207 ENN Clone1LC-CDR2 208 GTWDSSLSAGAV Clone1LC-CDR3 209 GFTFSNYA Clone1HC-CDR1 210 ISGSGGST Clone1HC-CDR2 211 ARPYYDD Clone1HC-CDR3 212 QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLIYENN KRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLSAGAVFGGG Clone1LCvariable TKLTVLG region 213 EVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSAI SGSGGSTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCARPYYD Clone1HCvariable DWGQGTLVTVSS region 214 HDIRNY Clone2LC-CDR1 215 AAS Clone2LC-CDR2 216 QQYDGLPLT Clone2LC-CDR3 217 ARYGSAAWMDS Clone2HC-CDR3 218 DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASN Clone2LCvariable LQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIK region R 219 QVQLVESGGGLVQPGGSLRLSCAASGFTFSNYAMSWVRQAPGKGLEWVSSI Clone2HCvariable SGSGGSTYYADSVKGRFTISRDTSKNTLYLQMNSLRAEDTAVYYCARYGSA region AWMDSWGQGTLVTVSS 220 GGGGS Linker 221 GGSG Linker 222 SGGG Linker 223 GSGS Linker 224 GSGSGS Linker 225 GSGSGSGS Linker 226 GSGSGSGSGS Linker 227 GGSGGS Linker 228 GGSGGSGGS Linker 229 GGSGGSGGSGGS Linker 230 GGSG Linker 231 GGSGGGSG Linker 232 GGSGGGSGGGSG Linker 233 SRGGGGSGGGGSGGGGSLEMA Linker 234 HHHHHH Histag 235 YPYDVPDYA HApeptide 236 YPYDVPDYAS HApeptide 237 DYKDDDDK FLAGpeptide 238 EQKLISEEDL Mycpeptide 239 QSVVTQPPSVSAAPGQKVTISCSGSSSNIGNNYVSWYQQLPGTAPKLLIYENN anti-CD22scFv KRPSGIPDRFSGSKSGTSATLGITGLQTGDEADYYCGTWDSSLSAGAVFGGG antibody TKLTVLGSRGGGGSGGGGSGGGGSLEMAEVQLVESGGGLVQPGGSLRLSCA ASGFTFSNYAMSWVRQAPGKGLEWVSAISGSGGSTYYADSVKGRFTISRDN SKNTLYLQMNSLRAEDTAVYYCARPYYDDWGQGTLVTVSSTSGQAGQHH HHHHGAYPYDVPDYAS 240 DIQLTQSPSSLSTSVGDRVTITCQASHDIRNYLNWYQQKPGKAPNLLIYAASN anti-CD22scFv LQTGVPSRFSGRGSGTDFTLTISSLQPEDIATYYCQQYDGLPLTFGQGTRLEIK antibody RSRGGGGSGGGGSGGGGSLEMAQVQLVESGGGLVQPGGSLRLSCAASGFTF SNYAMSWVRQAPGKGLEWVSSISGSGGSTYYADSVKGRFTISRDTSKNTLY LQMNSLRAEDTAVYYCARYGSAAWMDSWGQGTLVTVSSTSGQAGQHHHH HHGAYPYDVPDYAS

[0639] One or more features from any embodiments described herein or in the figures may be combined with one or more features of any other embodiment described herein in the figures without departing from the scope of the disclosure.

[0640] All publications, patents and patent applications cited in this specification are herein incorporated by reference as if each individual publication or patent application were specifically and individually indicated to be incorporated by reference. Although the foregoing disclosure has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be readily apparent to those of ordinary skill in the art in light of the teachings of this disclosure that certain changes and modifications may be made thereto without departing from the spirit or scope of the appended claims.