Ultra-potent neutralization of GM-CSF by multispecific antibodies and uses thereof

Abstract

The present invention provides multispecific antibodies, and antigen binding fragments thereof, that potently neutralize a cytokine and that may thus be useful in the prevention and/or treatment of inflammatory and/or autoimmune diseases. In particular, the present invention provides a multispecific antibody, or an antigen binding fragment thereof, comprising at least two different domains specifically binding to at least two different, non-overlapping sites in a cytokine and an Fc moiety. The invention also relates to nucleic acids that encode such antibodies and antibody fragments and immortalized B cells and cultured plasma cells that produce such antibodies and antibody fragments. In addition, the invention relates to the use of the antibodies and antibody fragments of the invention in screening methods as well as in the diagnosis, prophylaxis and treatment of inflammatory and/or autoimmune diseases.

Claims

1. A method for prophylaxis, treatment, or attenuation of an inflammatory and/or an autoimmune disease, comprising administering to the subject an effective amount of a multispecific anti-GM-CSF antibody, or an antigen binding fragment thereof, comprising: (a) at least two epitope binding sites, wherein each of the at least two epitope binding sites is specific for a different epitope of GM-CSF, whereby the different epitopes of the GM-CSF are non-overlapping epitopes; and (b) an Fc moiety, wherein the antibody or antigen binding fragment comprises: (i) CDRH1, CDRH2, and CDRH3 amino acid sequences and CDRL1, CDRL2, and CDRL3 amino acid sequences according to SEQ ID NOs: 1-5 and 7, or functional sequence variants thereof, respectively, or according to SEQ ID NOs: 1-4 and 6-7, or functional sequence variants thereof, respectively; (ii) CDRH1, CDRH2, and CDRH3 amino acid sequences and CDRL1, CDRL2, and CDRL3 amino acid sequences according to SEQ ID NOs: 49-53 and 55, or functional sequence variants thereof, respectively, or according to SEQ ID NOs: 49-52 and 54-55, or functional sequence variants thereof, respectively; (iii) CDRH1, CDRH2, and CDRH3 amino acid sequences and light chain CDRL1, CDRL2, and CDRL3 amino acid sequences according to SEQ ID NOs: 67-71 and 73, or functional sequence variants thereof, respectively, or according to SEQ ID NOs: 67-70 and 72-73, or functional sequence variants thereof, respectively; and/or (iv) heavy chain CDRH1, CDRH2, and CDRH3 amino acid sequences and light chain CDRL1, CDRL2, and CDRL3 amino acid sequences according to SEQ ID NOs: 105-109 and 111, or functional sequence variants thereof, respectively, or according to SEQ ID NOs: 105-108 and 110-111, or functional sequence variants thereof, respectively.

2. The method according to claim 1, wherein the antibody, or the antigen binding fragment thereof, is bispecific, trispecific, tetraspecific, or pentaspecific.

3. The method according to claim 1, wherein the at least two epitope bindings sites consist of two different epitope binding sites specific for two different, non-overlapping epitopes of the GM-CSF.

4. The method according to claim 2, wherein the antibody, or the antigen binding fragment thereof, is a bispecific tetravalent antibody or a trispecific hexavalent antibody.

5. The method according to claim 1, wherein the antibody, or the antigen binding fragment thereof, further comprises: (c) at least one linker.

6. The method according to claim 5, wherein the linker comprises or consists of: (i) the amino acid sequence according to SEQ ID NO: 143, or a functional sequence variant thereof; or (ii) the amino acid sequence according to SEQ ID NO: 144, or a functional sequence variant thereof.

7. The method according to claim 1, wherein the antibody, or the antigen binding fragment thereof, comprises: (a) an IgG type, (b) an IgG1 type, (c) a heavy chain constant region of the IgG1 CH1-CH2-CH3 type and a light chain constant region of the IgG CK type, or (d) a heavy chain constant region of the IgG1 CH1-CH2-CH3 type comprising or consisting of an amino acid sequence according to SEQ ID NO: 140 or functional sequence variants thereof, and a light chain constant region of the IgG CK type comprising or consisting of an amino acid sequence according to SEQ ID NO: 141 or functional sequence variants thereof.

8. The method according to claim 1, wherein the antibody, or the antigen binding fragment thereof, is of a construct type selected from the group consisting of Bs1, Bs2, Bs3, Ts1, Ts2 and Ts3.

9. The method according to claim 8, wherein the antibody, or the antigen binding fragment thereof, comprises a construct type Ts3 or comprises a trispecific antibody according to the construct type Ts3.

10. The method according to claim 1, wherein the antibody, or the antigen binding fragment thereof, is a human antibody, a monoclonal antibody, a human monoclonal antibody, a purified antibody, or a single chain antibody, or an antigen binding fragment thereof.

11. The method according to claim 1, wherein the antibody, or the antigen binding fragment thereof, neutralizes the GM-CSF: (i) under stringent conditions with an IC.sub.90 of 150 ng/ml or less; (ii) under less stringent conditions with an IC.sub.90 of 20 ng/ml or less; (iii) under more stringent conditions with an IC.sub.90 of 160 ng/ml or less; and/or (iv) under very stringent conditions with an IC.sub.90 of 1000 ng/ml or less.

12. The method according to claim 1, wherein the antibody or antigen binding fragment comprises: (i) a VH amino acid sequence according to SEQ ID NO: 37 or a functional sequence variant thereof and a VL amino acid sequence according to SEQ ID NO: 38 or a functional sequence variant thereof; (ii) a VH amino acid sequence according to SEQ ID NO: 63 or a functional sequence variant thereof and a VL amino acid sequence according to SEQ ID NO: 64 or a functional sequence variant thereof; (iii) a VH amino acid sequence according to SEQ ID NO: 95 or a functional sequence variant thereof and a VL amino acid sequence according to SEQ ID NO: 96 or a functional sequence variant thereof; and/or (iv) a VH amino acid sequence according to SEQ ID NO: 130 or a functional sequence variant thereof and a VL amino acid sequence according to SEQ ID NO: 131 or a functional sequence variant thereof.

13. The method according to claim 1, wherein the heavy chain of the antibody or antigen binding fragment comprises a VL amino acid sequence selected from the amino acid sequences according to SEQ ID NOs: 38, 64, 96, or 131, or functional sequence variants thereof; or the heavy chain of the antibody or antigen binding fragment comprises a VL amino acid sequence according to SEQ ID NOs: 38 or 96, or functional sequence variants thereof; or the heavy chain of the antibody or antigen binding fragment comprises a VL amino acid sequence according to SEQ ID NO: 96 or a functional sequence variant thereof.

14. The method according to claim 1, wherein the antibody or antigen binding fragment: (i) is of a construct type selected from the group consisting of the construct types Bs1, Bs2, Bs3, Ts1, Ts2 and Ts3; and (ii) comprises at any of the positions A and/or C a CDRH1 amino acid sequence, a CDRH2 amino acid sequence, a CDRH3 amino acid sequence, a CDRL1 amino acid sequence, a CDRL2 amino acid sequence and a CDRL3 amino acid sequence selected from the group consisting of amino acid sequences according to SEQ ID NOs: 1-7 and 67-73, or functional sequence variants thereof, respectively.

15. The method according to claim 1, wherein the antibody, or the antigen binding fragment thereof, is according to gTs1GC1, gTs1GC2a, gTs2GC2b, gTs2GC2c, gTs3GC2d, gTs3GC2e, gBs3GC1a, gBs3GC1b, gBs2GC1c, gBs2GC1d, gBs1GC2a, gBs3GC2b, gBs1GC3a, gBs3GC3b, gBs3GC4, or gBs3GC5.

16. The method according to claim 15, wherein the antibody, or the antigen binding fragment thereof, is Ts1GC1, Ts1GC2a, Ts2GC2b, Ts2GC2c, Ts3GC2d, Ts3GC2e, Bs3GC1a, Bs3GC1b, Bs2GC1c, Bs2GC1d, Bs1GC2a, Bs3GC2b, Bs1GC3a, Bs3GC3b, Bs3GC4, or Bs3GC5.

17. The method according to claim 1, wherein the inflammatory disease is due to a physical cause, an immune reaction due to hypersensitivity, a chemical cause, or is elected from the group consisting of appendicitis, bursitis, colitis, cystitis, dermatitis, phlebitis, RSD/CRPS, rhinitis, tendonitis, tonsillitis, vasculitis, Alzheimer's disease, ankylosing spondylitis, arthritis (osteoarthritis, rheumatoid arthritis (RA), psoriatic arthritis, asthma, atherosclerosis, Crohn's disease, colitis, dermatitis, diverticulitis, fibromyalgia, hepatitis, irritable bowel syndrome (IBS), systemic lupus erythematous (SLE), nephritis, Parkinson's disease, ulcerative colitis, Acne vulgaris, autoinflammatory diseases, celiac disease, prostatitis, pulmonary alveolar proteinosis, glomerulonephritis, hypersensitivities, inflammatory bowel diseases, pelvic inflammatory disease, reperfusion injury, sarcoidosis, transplant rejection, vasculitis, interstitial cystitis, inflammatory myopathies, encephalomyelitis, in particular acute disseminated encephalomyelitis, spondylitis, in particular ankylosing spondylitis, anti synthetase syndrome, dermatitis, in particular atopic dermatitis or contact dermatitis, hepatitis, in particular autoimmune hepatitis, autoimmune peripheral neuropathy, pancreatitis, in particular autoimmune pancreatitis, Behet's disease, Bickerstaffs, encephalitis, Blau syndrome, Coeliac disease, Chagas disease, polyneuropathy, in particular chronic inflammatory demyelinating polyneuropathy, osteomyelitis, in particular chronic recurrent multifocal osteomyelitis, Churg-Strauss syndrome, Cogan syndrome, giant-cell arteritis, CREST syndrome, vasculitis, in particular cutaneous small-vessel vasculitis and urticarial vasculitis, dermatitis herpetiformis, dermatomyositis, systemic scleroderma, Dressler's syndrome, drug-induced lupus erythematosus, discoid lupus erythematosus, enthesitis, eosinophilic fasciitis, eosinophilic gastroenteritis, erythema nodosum, Idiopathic pulmonary fibrosis, gastritis, Grave's disease, Guillain-barre syndrome, Hashimoto's thyroiditis, Henoch-Schonlein purpura, hydradenitis suppurativa, Idiopathic inflammatory demyelinating diseases, myositis, in particular inclusion body myositis, cystitis, in particular interstitial cystitis, Kawasaki disease, Lichen planus, lupoid hepatitis, Majeed syndrome, Meniere's disease, microscopic polyangitis, mixed connective tissue disease, myelitis, in particular neuromyelitis optica, thyroiditis, in particular Ord's thyroiditis, rheumatism, in particular palindromic rheumatism, Parsonage-Turner syndrome, pemphigus vulgaris, perivenous encephalomyelitis, polyarteritis nodosa, polymyalgia, in particular polymyalgia rheumatica, polymyositis, cirrhosis, in particular primary biliary cirrhosis, cholangitis, in particular primary sclerosing cholangitis, progressive inflammatory neuropathy, Rasmussen's encephalitis, relapsing polychondritis, arthritis, in particular reactive arthritis (Reiter disease) and rheumatoid arthritis, rheumatic fever, sarcoidosis, Schnitzler syndrome, serum sickness, spondyloarthropathy, Takayasu's arteritis, Tolosa-Hunt syndrome, transverse myelitis, or Wegener's granulomatosis.

18. The method of claim 1, wherein the autoimmune disease is a disease of the CNS, an auto-inflammatory disease, Celiac disease, Sjogren's syndrome, systemic lupus erythematosus, Blau syndrome, Bullous pemphigoid, Cancer, Castleman's disease, Celiac disease, Chagas disease, Chronic inflammatory demyelinating polyneuropathy, Chronic recurrent multifocal osteomyelitis, chronic obstructive pulmonary disease, Churg-Strauss syndrome, Cicatricial pemphigoid, Cogan syndrome, Cold agglutinin disease, Complement component 2 deficiency, Contact dermatitis, Cranial arteritis, CREST syndrome, Crohn's disease, Cushing's Syndrome, Dercum's disease, Dermatitis herpetiformis, Dermatomyositis, Diabetes mellitus type 1, Diffuse cutaneous systemic sclerosis, Dressler's syndrome, lupus, Discoid lupus erythematosus, Eczema, Acute disseminated encephalomyelitis (ADEM), Addison's disease, Agammaglobulinemia, Amyotrophic lateral sclerosis (Also Lou Gehrig's disease; Motor Neuron Disease), Ankylosing Spondylitis Antiphospholipid syndrome, Anti synthetase syndrome, Atopic dermatitis, Autoimmune aplastic anemia, Autoimmune cardiomyopathy, Autoimmune hemolytic anemia, Autoimmune hepatitis, Autoimmune inner ear disease, Autoimmune lymphoproliferative syndrome, Autoimmune peripheral neuropathy, Autoimmune pancreatitis, Autoimmune polyendocrine syndrome, Autoimmune progesterone dermatitis, Autoimmune thrombocytopenic purpura, Autoimmune urticarial, Autoimmune uveitis, Balo disease/Balo concentric sclerosis, Behcet's disease, Berger's disease, Bickerstaff s encephalitis, Endometriosis, Enthesitis-related arthritis, Eosinophilic gastroenteritis, Epidermolysis bullosa acquisita, Erythroblastosis fetalis, Evan's syndrome, Fibrodysplasia ossificans, Fibrosing alveolitis (or Idiopathic pulmonary fibrosis), Gastritis, Glomerulonephritis, Goodpasture's syndrome, Graves' disease, Guillain-Barre syndrome, Hashimoto's encephelopathy, Hashimoto's thyroiditis, Gestational Pemphigoid, Hidradenitis suppurativa, Hypogammaglobulinemia, Idiopathic thrombocytopenic purpura (Autoimmune thrombocytopenic purpura), IgA nephropathy, Occular cicatricial pemphigoid, Inclusion body myositis, Rheumatoid arthritis, Chronic inflammatory Rheumatic fever, demyelminating polyneuropathy, Sarcoidosis, Palindromic rheumatism, Interstitial cystitis, Juvenile idiopathic Schizophrenia, PANDAS (pediatric arthritis aka Juvenile autoimmune rheumatoid arthritis), Schmidt syndrome, neuropsychiatric Kawasaki's disease another form of APS, Schnitzler syndrome, Paraneoplastic cerebellar myasthenic syndrome, Leukocytoclastic Serum Sickness, Lichen planus, Sjogren's syndrome, Lichen sclerosus, Parsonage-Turner, Linear IgA disease, Still's disease, Pemphigus vulgaris, Lupoid hepatitis, Autoimmune hepatitis, Stiff person syndrome, Pernicious anaemia, Subacute bacterial endocarditis (SBE), POEMS syndrome, Lupus erythematosus, Sweet's syndrome, Sympathetic ophthalmia, Meniere's disease, Systemic lupus, Primary biliary cirrhosis, Miller-Fisher syndrome, Takayasu's arteritis, cholangitis, Progressive inflammatory neuropathy, Mucha-Habermann disease, Psoriasis, Psoriatic arthritis, Pyoderma gangrenosum, Multiple sclerosis, Pure red cell aplasia, Rasmussen's encephalitis, Myasthenia gravis, Transverse myelitis, Raynaud phenomenon, Microscopic colitis, Ulcerative colitis, Myositis, idiopathic inflammatory bowel disease (IBD), Neuromyelitis optica, Devic's disease, or Neuromyotonia.

19. The method according to claim 1, wherein the antibody or antigen binding fragment comprises: (i) CDRH1, CDRH2, and CDRH3 amino acid sequences and CDRL1, CDRL2, and CDRL3 amino acid sequences according to SEQ ID NOs: 1-5 and 7, respectively, or according to SEQ ID NOs: 1-4 and 6-7, respectively; (ii) CDRH1, CDRH2, and CDRH3 amino acid sequences and CDRL1, CDRL2, and CDRL3 amino acid sequences according to SEQ ID NOs: 49-53 and 55, respectively, or according to SEQ ID NOs: 49-52 and 54-55, respectively; (iii) CDRH1, CDRH2, and CDRH3 amino acid sequences and light chain CDRL1, CDRL2, and CDRL3 amino acid sequences according to SEQ ID NOs: 67-71 and 73, respectively, or according to SEQ ID NOs: 67-70 and 72-73, respectively; and/or (iv) heavy chain CDRH1, CDRH2, and CDRH3 amino acid sequences and light chain CDRL1, CDRL2, and CDRL3 amino acid sequences according to SEQ ID NOs: 105-109 and 111, respectively, or according to SEQ ID NOs: 105-108 and 110-111, respectively.

20. The method according to claim 12, wherein the antibody or antigen binding fragment comprises: the VH amino acid sequence according to SEQ ID NO: 37 and the VL amino acid sequence according to SEQ ID NO: 38; (ii) the VH amino acid sequence according to SEQ ID NO: 63 and the VL amino acid sequence according to SEQ ID NO: 64; (iii) the VH amino acid sequence according to SEQ ID NO: 95 and the VL amino acid sequence according to SEQ ID NO: 96; and/or (iv) the VH amino acid sequence according to SEQ ID NO: 130 and the VL amino acid sequence according to SEQ ID NO: 131.

21. The method according to claim 14, wherein the antibody or antigen binding fragment comprises at position A a CDRH1 amino acid sequence, a CDRH2 amino acid sequence, a CDRH3 amino acid sequence, a CDRL1 amino acid sequence, a CDRL2 amino acid sequence and a CDRL3 amino acid sequence according to SEQ ID NOs: 67-71 and 73, or functional sequence variants thereof, or according to SEQ ID NOs: 67-70 and 72-73, or functional sequence variants thereof.

22. The method according to claim 1, wherein the antibody or antigen binding fragment comprises: (i) a VH amino acid sequence that is at least 80% identical to SEQ ID NO: 37 and a VL amino acid sequence that is at least 80% identical to SEQ ID NO: 38; (ii) a VH amino acid sequence that is at least 80% identical to SEQ ID NO: 63 and a VL amino acid sequence that is at least 80% identical to SEQ ID NO: 64; (iii) a VH amino acid sequence that is at least 80% identical to SEQ ID NO: 95 and a VL amino acid sequence that is at least 80% identical to SEQ ID NO: 96; and/or (iv) a VH amino acid sequence that is at least 80% identical to SEQ ID NO: 130 and a VL amino acid sequence that is at least 80% identical to SEQ ID NO: 131.

23. The method according to claim 16, wherein the antibody, or the antigen binding fragment thereof, is Bs1GC3a.

24. The method according to claim 16, wherein the antibody, or the antigen binding fragment thereof, is Ts3GC2d.

Description

DESCRIPTION OF FIGURES

(1) FIG. 1 shows (A) SPR cross-competition between GCA21, GCA7 and GCB59, (B) A multichannel chip coated with GCA21, GCA7 or site IV GCB59 was saturated with GM-CSF and serially exposed to an excess of the same antibodies. (C) SEC-HPLC profile of samples containing the three non-cross-competing antibodies, alone or with GM-CSF added in equimolar concentrations (1:1) or in 10-fold antibody excess (10:1).

(2) FIG. 2 shows the potent in vitro neutralization of GM-CSF by a combination of three antibodies. A fixed amount of GM-CSF (final concentration 50 pg ml-1) was incubated with serial dilutions of one or more antibodies, added to TF1 cells (10,000 per well) and cell proliferation was measured on day 3 by thymidine incorporation. (A) Scheme of the TF-1 bioassay. (B) Serial dilutions of single monoclonal antibodies or mixtures of two and three non-cross-competing antibodies were tested for their capacity to neutralize GM-CSF. (C) The sensitivity of the test was changed by varying the number of cells and the concentration of GM-CSF as indicated. Shown is for each experimental condition the inhibition obtained using single antibodies or a combination of three non-cross-competing antibodies.

(3) FIG. 3 shows the Fc-dependent clearance of GM-CSF immune complexes in vivo. (A) A sandwich ELISA to detect GM-CSF in the presence of specific antibodies. A fixed amount of GM-CSF was added to mouse serum together with three monoclonal antibodies (GCA21, GCA7, GCB59) added separately or in combination. The quantification of GM-CSF was performed by a sandwich ELISA using an antibody specific for site II for capture and site I for detection. Serial dilutions of serum in neutral (left) or alkaline buffer (right) were added and GM-CSF concentration was determined with reference to a GM-CSF standard. The dotted line represents the concentration of GM-CSF measured in the absence of antibodies. (B) Female Balb/c mice (5 per group) were injected with 100 μg of monoclonal antibody, either GCA21 (1 mAb) or GCA21+GCA7+GCB59 (3 mAbs) in the IgG or IgG-LALA format, or with 2 mg total IgG from a PAP patient, followed by 2 μg GM-CSF after 16 hours. Sera were collected after 1 or 5 days and GM-CSF concentrations were measured by ELISA in untreated serum and in serum treated at pH 11.6 to dissociate immune complexes. Shown is the GM-CSF concentration on day 1 and on day 5 in untreated serum (left) or alkaline-treated serum (right). (C) Proliferation of TF-1 cells in response to different dilutions of serum of mice injected 24 hours before with GM-CSF and the indicated antibodies. (D) Binding of GM-CSF immune complexes formed by one or three antibodies (in the IgG1 or IgG1-LALA format) to TZM-b1 cells expressing Fc□RIIa or Fc□□RIIb, as measured by flow cytometry using an anti-IgG Fc specific antibody.

(4) FIG. 4 shows the scheme of three bispecific construct types Bs1, Bs2 and Bs3 and of the three trispecific construct types Ts1, Ts2 and Ts3. Positions A, B, and/or C (if applicable) are shown for each of the construct types. Single chain variable domains (ScVd) of both heavy chain and light chain of different GM-CSF antibodies are added at the N-terminus and/or at the C-terminus of the heavy chain or the light chain of one GM-CSF antibody used as scaffold. Black ovals represent VH domains while dark grey ovals represent VL domains. The VH and VL of the different ScVds are joined together through specific linkers. Other linkers are used to join the ScVds between one another and to the full antibody used as scaffold. Light gray ovals represent IgG1 CH and CL domains.

(5) FIG. 5 shows that TsGC1 binds to GM-CSF with very high affinity with a very slow off-rate (A). (B-D) TsGC1 can use all the 3 different specificities for binding to GM-CSF. GCA21, GCA7 and GCB59 were immobilized on a SPR chip and were serially exposed to GM-CSF followed by GCA7, GCB59 and GCA21, respectively, and finally by TsGC1. (E) TsGC1 can form high molecular weight complexes with GM-CSF. TsGC1 was immobilized on an SPR chip and serially exposed for 3 rounds to GM-CSF followed by soluble TsGC1. Shown is the same experiment performed with GCA7 as control.

(6) FIG. 6 shows the extremely potent neutralization of GM-CSF by TsgC2d and BsGC3a as compared to single antibodies or combinations of antibodies forming TsgC2d and BsGC3a.

(7) FIG. 7 shows the binding of immune complexes formed by GM-CSF and TsGC2d or BsGC3a to TZM-b1 cells expressing FcγRIIa or FcγRIIb, as measured by flow cytometry using an anti-IgG Fc specific antibody. The binding is compared to that of single antibodies and combinations of the 2 or 3 antibodies forming BsGC3a and TsGC2d, respectively.

EXAMPLES

(8) Exemplary embodiments of the present invention are provided in the following examples. The following examples are presented only by way of illustration and to assist one of ordinary skill in using the invention. The examples are not intended in any way to otherwise limit the scope of the invention.

Example 1: Isolation and Characterization GM-CSF-Specific Antibodies

(9) Peripheral blood samples were collected from five pulmonary alveolar proteinosis (PAP) patients. IgG memory B cells were isolated from cryopreserved or fresh PBMCs by a combination of magnetic and fluorescence-activated cell sorting, in particular using anti-FITC microbeads (Miltenyi Biotec) following staining of PBMCs with CD22-FITC (BD Phamingen). IgG memory B cells were then immortalized in clonal conditions with EBV (Epstein-Barr virus) and CpG in 384 well micro-plates in the presence of feeder cells as described by Traggiai E. et al. (2004) Nat Med. 10(8):871-5 and WO 2004/076677 A2. The culture supernatants were screened for the presence of GM-CSF-specific IgG antibodies by ELISA. Four immortalized B cell clones that produced GM-CSF monoclonal antibodies were identified. cDNA was synthesized from positive cultures and the antibody V genes (heavy chain and light chain variable regions) were sequenced and analyzed using the IMGT database (http://www.imgt.org/). The V(D)J gene usage of the four PAP autoantibodies is shown in Table 5.

(10) TABLE-US-00005 TABLE 5 V(D)J gene usage of the 4 PAP autoantibodies. The antibodies use different V, D and J genes and are somatically mutated. The load of somatic mutations was comparable to that characteristic of T-cell-dependent responses against non-self-antigens, ranging from 8.8% to 16.7% in the VH gene segment and from 0% to 8.6% in the VL gene segment. mAb Heavy chain VDJ genes (% identity to GL) Light chain VJ genes (% identity to GL) GCA7 VH3-66 (91.2) D3-10 JH4 (93.8) VK4-1 (96)   JK3 (100)   GCA21 FH3-30-3 (83.3) D2-15 JH2 (84.9) VK1-5 (92.8) JK4 (97.2) GCB59 FH3-21 (86.8) D2-15 JH6 (77.4) VL3-21 (92.1) JL2 (91.9) GCE536 FH1-46 (87.9) D2-2 JH6 (85.5) FK3-20 (91.5) JK2 (92.1)

(11) All antibodies were recombinantly produced as IgG1 by transient transfection of HEK 293 Freestyle Cells (Invitrogen) using polyethylenimine (PEI). The antibodies were then tested for binding to human GM-CSF by ELISA. Binding properties and V-gene usage is shown in Table 5. The antibodies showed high affinities comparable to the antibodies MOR103 and namilumab, which are GM-CSF neutralizing monoclonal antibodies under clinical development, which serve herein as reference antibodies. The EC50 values (ng/ml) were determined by ELISA and calculated for every sample by nonlinear regression analysis using GraphPad Prism 5 software. The EC50 values ranged from 61.4 to 307.6 ng/ml. Interestingly, the antibodies did not cross-react with both mouse and rat GM-CSF.

(12) The kinetics of binding was determined by surface plasmon resonance (SPR). Briefly, for SPR Protein A (450 nM) was stabilized in 10 mM acetate buffer, pH 4.5, and immobilized onto a EDC/NHS pre-activated ProteOn sensor chip (Biorad) through amine coupling; unreacted groups were blocked by injection of ethanolamine HCl (1 M). HEPES buffered saline (HBS) (10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.005% surfactant Tween-20) was used as running buffer. All injections were made at flow rate of 100 □l/min. Monoclonal antibodies were diluted in HBS (200 nM) and injected onto the protein A coated chip for capturing, followed by injection of different concentrations of human GM-CSF (400 nM, 200 nM, 100 nM, 50 nM, 25 nM); one channel of the chip was injected with HBS and used as reference for the analysis. Injection time and dissociation time were 120 s and 600 s, respectively. Each binding interaction of mAbs with GM-CSF was assessed using a ProteON XPR36 instrument (Biorad) and data processed with ProteOn Manager Software. Ka, Kd and KD were calculated applying the Langmuir fit model. The KD determined ranged from 0.18 to 0.69 nM, consistent with the high affinity binding. However, the kinetic values were highly heterogeneous. For instance, antibodies GCA7 and GCB59 had comparable KD values (0.38 and 0.68 nM, respectively), but showed different kinetics with GCA7 being characterized by a slow on-/slow off-rate and GCB59 by a high on-/high off-rate (Table 6).

(13) TABLE-US-00006 TABLE 6 Binding properties and V-gene usage of GM-CSF autoantibodies from PAP patients. Human monoclonal antibodies from PAP patients show high affinities comparable to reference monoclonal antibodies. The table shows EC50 values determined by ELISA and Ka, Kd and KD values determined by SPR; reference monoclonal antibodies are highlighted in bold. mAb EC50 (ng/ml) Ka (1/Ms) Kd (1/s) KD (M) GCA7 186.8 2.4E+05 6.0E−05 3.8E−10 GCA21 59.4 9.5E+05 6.5E−04 6.9E−10 GCB59 307.6 1.7E+06 1.2E−03 6.8E−10 GCE536 61.4 6.6E+05 1.1E−04 1.8E−10 Clone 3092 61.0 5.9E+05 3.4E−04 5.7E−10 Clone 1089 1080.0 1.8E+05 7.9E−05 4.4E−10 MOR103 90.0 2.7E+05 1.5E−05 1.9E−10 Namilumab 75.3 3.1E+05 7.7E−05 2.4E−10

Example 2: The Four PAP Autoantibodies Recognize Distinct Sites on GM-CSF and can Form High-Molecular-Weight Immune Complexes

(14) To assess simultaneous binding of the different mABs to GM-CSF, SPR cross competition experiments were performed by SPR as described above (Example 1) whereby the different mAbs (200 nM each) were serially injected after GM-CSF capture (50 nM). Injection time and dissociation time were 60 s and 20 s, respectively. Thereby, it was found that GCA7, GCA21, GCB59 and GCE536 do not cross-compete between them for binding to GM-CSF (FIG. 1A). Interestingly, SPR experiments show that three non-cross-competing autoantibodies can bind simultaneously to a single molecule of GM-CSF (FIG. 1B). Furthermore, when GM-CSF was incubated with an excess of three antibodies, formation of high-molecular-weight immune complexes could be detected by size-exclusion chromatography (SEC-HPLC) (FIG. 1C). In the SEC-HPLC experiment three non-cross competing mAbs were diluted in PBS singularly or as a three-antibody-combination (10 □g of total antibody amount), and mixed with GM-CSF (1:1 or 10:1 molar ratios) for 1 hour, RT. Samples were analyzed by Agilent 1100 HPLC machine using TSK-GEL G3000SW columns (Tosoh, bed volume: 13 ml, void volume: 4.6 ml) with PBS as mobile phase (flow rate: 1 ml/min). A universal solvent 2 □m filter (Agilent) was put between injector and column. Detection was performed by a Variable Wavelength Detector (VWD, Agilent) with ultraviolet absorption at 220 nm.

Example 3: Potent In Vitro Neutralization of GM-CSF Requires Combinations of 3 Antibodies Binding to Non-Overlapping Sites

(15) The neutralizing activity of the autoantibodies was assessed by measuring their ability to inhibit the proliferation of TF-1 cells in response to recombinant GM-CSF (FIG. 2A). To this end, TF-1 cells (CLS, Cell Lines Service) were maintained in RPMI 1640 medium supplemented with 10% Fetal Bovine Serum (Hyclone), 1% GlutaMAX, 1% Penicillin/Streptavidin, 1% non-essential amino acids, 1% sodium pyruvate, 1‰ 2-mercaptoethanol (all from GIBCO), 5 ng/ml human GM-CSF (Gentaur), 10 ng/ml human IL-3 (ImmunoTools). Cells were grown at 37° C. in a humidified incubator with 5% CO2. A GM-CSF neutralization assay was performed by serially diluting mAbs (or combination of mAbs, total IgG, or affinity-purified antibodies) in growth medium with neither GM-CSF nor IL-3, adding GM-CSF at a concentration of 100 pg/ml, and preincubating in 96-well flat-bottom cell culture plates (Costar) at 37° C. for 1 hour. TF-1 cells were washed 5 times, diluted in growth medium with neither GM-CSF nor IL-3, and 10,000 cells per well were seeded (final GM-CSF concentration equal to 50 pg/ml). In other tests, GM-CSF was used at final concentration of 500 and 5,000 pg/ml, and 1,000 cells per wells were seeded. Cells with or without GM-CSF in absence of antibodies were used as control to determine maximum and minimum levels of cell proliferation. Plates were incubated at 37° C. in a humidified incubator with 5% CO2 for 72 hours, and cell proliferation was measured after 6-hour incubation with 0.2 □Ci/well of [3H]-thymidine (PerkinElmer). GM-CSF neutralization was calculated as percentage of inhibition of TF-1 growth with the following formula: [1-(CCPM of a single well—average CCPM of control cells grown without GM-CSF)/(average CCPM of control cells grown with GM-CSF−average CCPM of control cells grown without GM-CSF)]×100 (CCPM=corrected counts per minute). IC.sub.90 (□g/ml) was calculated for every sample by a nonlinear regression analysis using GraphPad Prism 5 software. In some experiments mouse sera were titrated in TF-1 growth medium and preincubated at 37° C. for 30 min. TF-1 cells were washed and seeded (1,000 cells per well). A titration of GM-CSF (60,000 to 0.3 ng/ml) was added as growth control. CCPM of each single well were plotted against the serum titration.

(16) Surprisingly, using this bioassay, GCA21, GCA7 and GCB59 failed to neutralize GM-CSF (FIG. 2B), even when tested at the concentration of 1 mg/ml (data not shown). The only exception was GCE536 which neutralized GM-CSF activity with an IC.sub.90 value of 2.43 μg/ml, while the therapeutic antibodies Namilumab and MOR103 showed IC.sub.90 values of 0.80 and 0.16, respectively. Interestingly, when combined together, two non-cross-competing antibodies showed enhanced neutralizing activity both in terms of dose-response and percent inhibition, the combination of GCA21 and GCB59 being the most effective (FIG. 2B). Strikingly, a combination of three non-cross-competing antibodies (GCA21, GCA7 and GCB59) led to a complete inhibition of proliferation with an IC.sub.90 value of 0.08 μg/ml (expressed as the total concentration of the three mAbs), which was lower than that of the therapeutic antibodies MOR103 and Namilumab (FIG. 2B).

(17) As expected from the law of mass action, it was found that by varying the cell number and the GM-CSF concentration the sensitivity of the assay was dramatically affected. In particular, lowering the number of TF-1 cells and the concentration of GM-CSF led to a more sensitive test that showed increased neutralization by single and multiple antibodies (FIG. 2C). In contrast, when high number of TF-1 cells and high doses of GM-CSF were used, even the most potent neutralizing antibodies MOR103 and Namilumab, failed to neutralize GM-CSF, even when present in a 400-fold molar excess. Strikingly, in all conditions, a combination of three non-cross-competing antibodies was capable of completely neutralizing GM-CSF (FIG. 2C).

Example 4: FcR-Dependent Clearance of GM-CSF Immune Complexes In Vivo

(18) Having established that GM-CSF can form complexes with three antibodies resulting in efficient in vitro neutralization of the cytokine biological activity, the effect of single versus multiple autoantibodies in vivo was investigated. To this end, groups of 6-8 week-old female BALB/c mice were injected intravenously with 100 □g of purified mAbs or 2 mg of total IgG purified from PA96 patient. After 16 hours, 2 □g of human GM-CSF were injected. Sera samples were collected on day 1 and day 5. GM-CSF was quantified by a sandwich ELISA. Briefly, 10 □g/ml of an antibody that bound to site II of GM-CSF was used to coat 96-well Maxisorp plates (Nunc), which were then blocked with PBS+10% FBS (Gibco). All sera and GM-CSF, which was used as standard (range 3.4-600,000 pg/ml), were titrated and tested in parallel under different conditions (either untreated or after alkaline treatment to dissociate the immune complexes; FIG. 3A): in one plate all samples were supplemented with 25% (vol/vol) of an alkaline dissociation buffer (2.5% Triton X100, 2 M ethanolamine, 0.15 M NaCl, pH 11.6), in the other plate all samples were supplemented with 25% (vol/vol) of PBS+10% FBS. Plates were left overnight at RT. Detection of captured GM-CSF was made with 1 □g/ml of a biotinylated antibody that bound to site I of GM-CSF for 1 h, RT, followed by binding of 0.5 □g/ml streptavidin-AP (Jackson ImmunoResearch) for 1 h, RT. Plates were then washed, substrate (p-NPP, Sigma) was added and plates were read at 405 nm.

(19) In the absence of antibodies, the injected GM-CSF disappeared rapidly from the serum and was undetectable after 24 hours (FIG. 3B). In contrast, when single antibodies (GCA21 or MOR103) were used, high levels of GM-CSF were recovered from serum on day 1 and were still present on day 5. Of note, GM-CSF detection required alkaline dissociation in the case of MOR103 but not for GCA21, consistent with the different dissociation rates of the two antibodies (Table 6). In striking contrast, when mice received three non-cross-competing antibodies (GCA21, GCA7 and GCB59) or PAP IgG, GM-CSF was rapidly cleared since only low or undetectable amounts of the cytokine could be detected in the day-1 and day-5 sera, respectively, after alkaline dissociation.

(20) To address the possible role of Fc receptors in the clearance of GM-CSF, the same antibodies were tested in a variant form, called LALA, which does not bind to C1q or to Fc-γ receptors. Similarly to the wild-type antibodies, single LALA antibodies led to an increase in GM-CSF levels in serum. However, in contrast to what was observed for three wild-type antibodies, three LALA antibodies failed to clear GM-CSF, which was quantitatively recovered in the sera following alkaline dissociation even on day 5 (FIG. 3B).

(21) To ask whether the antibody-bound GM-CSF would be bioavailable, the sera of mice were tested for their ability to support TF-1 proliferation (FIG. 3C). Sera of mice receiving GCA21 or MOR103 led to a robust proliferation of TF-1 cells, consistent with a GM-CSF dissociation rate sufficient to engage the cytokine receptor. In contrast, sera of mice receiving three wild-type antibodies or PAP IgG were not able to stimulate proliferation, consistent with clearance of the immune complexes in vivo. In addition, although containing high level of GM-CSF, sera of mice receiving three LALA antibodies were not stimulatory, a finding consistent with irreversible sequestration of GM-CSF in stable immune complexes.

(22) To further address the role of Fcγ-receptors, immune complexes formed between GM-CSF and wild-type or LALA antibodies were tested for their capacity to bind to TZM-b1 cells expressing different Fcγ-receptors. To this end, four TZM-b1 cell lines (NIH AIDS Research & Reference Reagent Program) each transfected with a specific Fcγ receptor (FcγRI, FcγRIIa, FcγRIIb or FcγRIIIa) were maintained in DMEM medium supplemented with 10% Fetal Bovine Serum (Hyclone), 0.025 M Hepes, 10 □g/ml Gentamicin and 20 □g/ml Blasticidin. Untransfected TZM-b1 cells were used as negative control and were maintained in DMEM medium supplemented with 10% Fetal Bovine Serum (Hyclone) and 2% Penicillin/Streptavidin. Cells were grown at 37° C. in a humidified incubator with 5% CO2. Expression of specific FcγRs was assessed by staining TZM-b1 cells with FITC-conjugated anti-CD64 (anti-FcγRI), anti-CD32 (anti-FcγRIIa and anti-FcγRIIb) and anti-CD16 (anti-FcγRIIIa) antibodies (all from BD Pharmingen). Untrasfected and transfected TZM-b1 cells were washed with staining buffer (PBS with 10% Fetal Bovine Serum and 2 mM EDTA) and seeded in 96-well-plates at a density of 50,000 cells per well. A single anti-GM-CSF mAb (GCA21) or a combination of three non-cross-competing mAbs (GCA21, GCA7 and GCB59) at final concentration of 2.5 □g/ml were mixed with 0.05 □g/ml GM-CSF, or staining buffer (PBS with 10% Fetal Bovine Serum and 2 mM EDTA). The LALA versions of all antibodies and a mAb with a different specificity were included as controls. Samples were incubated at 37° C. for 30 min to allow the formation of immune complexes and then cooled down to 4° C. before adding them to TZM-b1 cells for 30 min. Cells were washed twice and stained with anti-human IgG Fcγ fragment specific F(ab′)2 fragment (Jackson ImmunoResearch). Samples were analyzed on BD FACSCanto (BD Biosciences) and median intensity fluorescence was analyzed and compared between samples.

(23) Strong binding was observed only on FcγRIIa- and FcγRIIb-expressing cells and when immune complexes were formed by three wild-type, but not LALA, antibodies (FIG. 3D). Taken together, the above results indicate that single antibodies, even when potently neutralizing in vitro, increase the half-life of GM-CSF and build up a circulating pool of bioavailable cytokine. In contrast, three or more antibodies lead to the formation of immune complexes that are efficiently cleared through an Fc-dependent mechanism.

Example 5: Engineering of Multispecific Antibodies with the Highest GM-CSF Neutralizing Activity

(24) In view of the need of targeting multiple independent sites on GM-CSF to achieve efficient in vivo neutralization and clearance of the cytokine, bispecific and trispecific antibodies carrying the GCA21, GCA7, GCB59 and GCE536 epitope binding sites, specific linkers and an intact human IgG1 Fc were designed and produced (Table 7). Six different construct types were used to produce the multispecific antibodies (FIG. 4). In particular the three construct types Ts1, Ts2 and Ts3 were used to produce 6 different trispecific hexavalent antibodies, while the three construct types Bs1, Bs2 and Bs3 were used to produce 10 different bispecific tetravalent antibodies (FIG. 4 and Table 7). The construct types Bs1, Bs2, Bs3, Ts1 and Ts2 were designed according to US 2009/0155275 A1.

(25) TABLE-US-00007 TABLE 7 Description of GM-CSF multispecific antibodies Ts1GC1 trispecific antibody Construct type Ts1 Specificities.sup.1 GCA21 + GCA7 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCA21 VH 63 65 chain.sup.2 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCA7 VH 37 42 Long linker 144 150 GCA7 VL 38 47 Short linker 143 149 GCB59 VH 95 100 Long linker 144 150 GCB59 VL 96 104 Complete sequence 151 152 Light GCA21 VL 64 66 chain.sup.2 IgG CK 141 147 Complete sequence 183 184 Ts1GC2a trispecific antibody Construct type Ts1 Specificities.sup.1 GCE536 + GCA7 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCE536 VH 130 132 chain.sup.2 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCA7 VH 37 42 Long linker 144 150 GCA7 VL 38 47 Short linker 143 149 GCB59 VH 95 100 Long linker 144 150 GCB59 VL 96 104 Complete sequence 153 154 Light GCE536 VL 131 136 chain.sup.2 IgG CK 141 147 Complete sequence 189 190 Ts2GC2b trispecific antibody Construct type Ts2 Specificities.sup.1 GCE536 + GCA7 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCB59 VH 95 98 chain.sup.2 Long linker 144 150 GCB59 VL 96 102 Short linker 143 149 GCE536 VH 130 133 IgG1 CH1—CH2—CH3 140 145 Complete sequence 155 156 Light GCA7 VH 37 41 chain.sup.2 Long linker 144 150 GCA7 VL 38 46 Short linker 143 149 GCE536 VL 131 137 IgG CK 141 147 Complete sequence 157 158 Ts2GC2c trispecific antibody Construct type Ts2 Specificities.sup.1 GCE536 + GCA7 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCB59 VH 95 99 chain.sup.2 Long linker 144 150 GCB59 VL 96 103 Short linker 143 149 GCA7 VH 37 40 IgG1 CH1—CH2—CH3 140 145 Complete sequence 159 160 Light GCE536 VH 130 134 chain.sup.2 Long linker 144 150 GCE536 VL 131 138 Short linker 143 149 GCA7 VL 38 45 IgG CK 141 147 Complete sequence 161 162 Ts3GC2d trispecific antibody Construct type Ts3 Specificities.sup.1 GCE536 + GCA7 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCB59 VH 95 98 chain.sup.2 Long linker 144 150 GCB59 VL 96 102 Short linker 143 149 GCE536 VH 130 133 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCA7 VH 37 43 Long linker 144 150 GCA7 VL 38 48 Complete sequence 163 164 Light GCE536 VL 131 136 chain.sup.2 IgG CK 141 147 Complete sequence 189 190 Ts3GC2e trispecific antibody Construct type Ts3 Specificities.sup.1 GCE536 + GCA7 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCB59 VH 95 99 chain.sup.2 Long linker 144 150 GCB59 VL 96 103 Short linker 143 149 GCA7 VH 37 40 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCE536 VH 130 135 Long linker 144 150 GCE536 VL 131 139 Complete sequence 165 166 Light GCA7 VL 38 44 chain.sup.2 IgG CK 141 147 Complete sequence 181 182 Bs3GC1a bispecific antibody Construct type Bs3 Specificities.sup.1 GCE536 + GCA7 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCE536 VH 130 132 chain.sup.2 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCA7 VH 37 43 Long linker 144 150 GCA7 VL 38 48 Complete sequence 167 168 Light GCE536 VL 131 136 chain.sup.2 IgG CK 141 147 Complete sequence 189 190 Bs3GC1b bispecific antibody Construct type Bs3 Specificities.sup.1 GCE536 + GCA7 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCA7 VH 37 39 chain.sup.2 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCE536 VH 130 135 Long linker 144 150 GCE536 VL 131 139 Complete sequence 169 170 Light GCA7 VL 38 44 chain.sup.2 IgG CK 141 147 Complete sequence 181 182 Bs2GC1c bispecific antibody Construct type Bs2 Specificities.sup.1 GCE536 + GCA7 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCE536 VH 130 132 chain.sup.2 IgG1 CH1—CH2—CH3 140 145 Complete sequence 187 188 Light GCA7 VH 37 41 chain.sup.2 Long linker 144 150 GCA7 VL 38 46 Short linker 143 149 GCE536 VL 131 137 IgG CK 141 147 Complete sequence 157 158 Bs2GC1d bispecific antibody Construct type Bs2 Specificities.sup.1 GCE536 + GCA7 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCA7 VH 37 39 chain.sup.2 IgG1 CH1—CH2—CH3 140 145 Complete sequence 179 180 Light GCE536 VH 130 134 chain.sup.2 Long linker 144 150 GCE536 VL 131 138 Short linker 143 149 GCA7 VL 38 45 IgG CK 141 147 Complete sequence 161 162 Bs1GC2a bispecific antibody Construct type Bs1 Specificities.sup.1 GCE536 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCB59 VH 95 98 chain.sup.2 Long linker 144 150 GCB59 VL 96 102 Short linker 143 149 GCE536 VH 130 133 IgG1 CH1—CH2—CH3 140 145 Complete sequence 155 156 Light GCE536 VL 131 136 chain.sup.2 IgG CK 141 147 Complete sequence 189 190 Bs3GC2b bispecific antibody Construct type Bs3 Specificities.sup.1 GCE536 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCB59 VH 95 97 chain.sup.2 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCE536 VH 130 135 Long linker 144 150 GCE536 VL 131 139 Complete sequence 171 172 Light GCB59 VL 96 101 chain.sup.2 IgG CL 142 148 Complete sequence 185 186 Bs1GC3a bispecific antibody Construct type Bs1 Specificities.sup.1 GCA7 + GCB59 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCB59 VH 95 99 chain.sup.2 Long linker 144 150 GCB59 VL 96 103 Short linker 143 149 GCA7 VH 37 40 IgG1 CH1—CH2—CH3 140 145 Complete sequence 159 160 Light GCA7 VL 38 44 chain.sup.2 IgG CK 141 147 Complete sequence 181 182 Bs3GC3b bispecific antibody Construct type Bs3 Specificities.sup.1 GCA7 + GCB59 Heavy Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl chain.sup.2 GCB59 VH 95 97 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCA7 VH 37 43 Long linker 144 150 GCA7 VL 38 48 Complete sequence 173 174 Light GCB59 VL 96 101 chain.sup.2 IgG CL 142 148 Complete sequence 185 186 Bs3GC4 bispecific antibody Construct type Bs3 Specificities.sup.1 GCA21 + GCE536 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCA21 VH 63 65 chain.sup.2 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCE536 VH 130 135 Long linker 144 150 GCE536 VL 131 139 Complete sequence 175 176 Light GCA21 VL 64 66 chain.sup.2 IgG CK 141 147 Complete sequence 183 184 Bs3GC5 bispecific antibody Construct type Bs3 Specificities.sup.1 GCA21 + GCA7 Domain/Linker SEQ ID NO. aa SEQ ID NO. nucl Heavy GCA21 VH 63 65 chain.sup.2 IgG1 CH1—CH2—CH3 140 146 Short linker 143 149 GCA7 VH 37 43 Long linker 144 150 GCA7 VL 38 48 Complete sequence 177 178 Light GCA21 VL 64 66 chain.sup.2 IgG CK 141 147 Complete sequence 183 184 .sup.1The antibody used as scaffold is underlined. .sup.2From N-terminus to C-terminus.

Example 6: Evaluation of Productivity and Aggregation of Multispecific Antibodies

(26) The anti GM-CSF multispecific antibodies were produced in 293F cells and purified on protein A. Quantification was performed by Pierce bicinchoninic acid (BCA) protein assay according to the manufacturer's instructions (Thermo Scientific). The assay is a detergent-compatible formulation based on BCA for the colorimetric detection and quantitation of total protein. Productivity varied according to the different antibodies with Bs3GC1a, Bs1GC2a, Bs1GC3a, Bs3GC3b, Bs3GC5 being produced at a concentration greater than 30 μg/ml. Aggregation of multispecific antibodies (final concentration: 1 mg/ml) was analyzed by measuring their turbidity at OD 340 nm in absence or presence of GM-CSF (final concentration: 0.1 mg/ml) (Table 8).

(27) TABLE-US-00008 TABLE 8 Productivity (μg/ml) of multispecific mAbs by 293F cells and their aggregation measured by turbidity at OD 340 nm in absence or presence of GM-CSF. Low OD values indicates lower level of turbidity. Turbidity Concentration Turbidity (OD 340) Name (μg/ml) (OD 340) w/GM-CSF Ts1GC1 3.2 0.345 0.369 Ts1GC2a 3 0.292 0.424 Ts2GC2b 7.2 0.076 0.169 Ts2GC2c 7.7 0.068 0.209 Ts3GC2d 13.5 0.012 0.197 Ts3GC2e 11.1 0.108 0.246 Bs3GC1a 30.6 0.084 0.121 Bs3GC1b 17.5 0.051 0.093 Bs2GC1c 7 0.058 0.133 Bs2GC1d 5 0 0.025 Bs1GC2a 41.8 0 0 Bs3GC2b 26 0.044 0.067 Bs1GC3a 61.5 0 0 Bs3GC3b 31.7 0 0 Bs3GC4 20.9 0.553 0.396 Bs3GC5 42 0.224 0.245

Example 7: Multispecific Antibodies Bind to GM-CSF with High Affinity

(28) All multispecific antibodies were tested for binding to GM-CSF by ELISA. The binding was highly specific with a high affinity for GM-CSF as shown by EC50 values in Table 9 below. The use of the different binding sites of the multispecific antibodies was tested by SPR experiments using Ts1GC1 as model. In this experiment GM-CSF was bound by an excess of 2 of the 3 antibodies forming Ts1GC1 and subsequent binding of Ts1GC1 to GM-CSF through the 3.sup.rd specificity was revealed. Ts1GC1 has a very high affinity, with very low KD as shown in FIG. 5A. In different SPR experiments GM-CSF was complexed with 2 antibodies out of the 3 antibodies composing Ts1GC1, so that only one of the 3 GM-CSF epitopes was left free for binding by Ts1GC1 (FIG. 5B-D). In addition, when immobilized on the SPR chip, Ts1GC1, unlike conventional antibodies such as GCA7, was able to form high molecular weight complexes when the chip was sequentially exposed to multiple rounds of soluble GM-CSF and Ts1GC1 (FIG. 5E).

(29) TABLE-US-00009 TABLE 9 Binding of multispecific mAbs to GM-CSF as determined by ELISA. Name EC50 (ng/ml) Ts1GC1 27.04 Ts1GC2a 33.48 Ts2GC2b 18.38 Ts2GC2c 20.71 Ts3GC2d 29.55 Ts3GC2e 40.67 Bs3GC1a 74.33 Bs3GC1b 71.6 Bs2GC1c 12 Bs2GC1d 22.56 Bs1GC2a 16.19 Bs3GC2b 48.55 Bs1GC3a 46.8 Bs3GC3b 586.7 Bs3GC4 32.27 Bs3GC5 41.75

Example 8: Extremely Potent Neutralization of GM-CSF by Multispecific Antibodies

(30) GM-CSF neutralization was tested using the in vitro bioassay based on TF-1 cells as described above (Example 3), whereby 2 different GM-CSF concentrations (50 and 500 pg/ml) and 2 different number of cells per well (1,000 and 10,000) were tested. IC90 values are reported in Table 10. Interestingly, all multispecific antibodies completely inhibited TF-1 proliferation in all conditions tested at very low concentrations (lower than 1 ng/ml for most multispecific antibodies). Of note, using less stringent conditions MOR103 and Namilumab required a 100-fold and 690-fold greater concentration, respectively, as compared to the best multispecific antibody (Ts1GC2a). Under stringent conditions, most multispecific antibodies could neutralize GM-CSF at concentrations lower than 10 ng/ml, while MOR103 and Namilumab required concentrations greater than 1 mg/ml. Two multispecific antibodies, Ts3GC2d and Bs1GC3a, were selected for their overall properties and compared to the single antibodies or combinations of antibodies from which they derived (FIG. 6).

(31) TABLE-US-00010 TABLE 10 Extremely potent neutralization of GM-CSF by multispecific antibodies. Less stringent conditions: 50 pg/ml GM-CSF and 1,000 TF-1/well. Stringent conditions: 50 pg/ml GM- CSF and 10,000 TF-1/well. More stringent conditions: 500 pg/ml GM-CSF and 1,000 TF-1/well. Very stringent conditions: 500 pg/ml GM-CSF and 10,000 TF-1/well IC90 (ng/ml) IC90 (ng/ml) IC90 (ng/ml) less IC90 (ng/ml) more very stringent stringent stringent stringent Name conditions conditions conditions conditions Ts1GC1 0.3516 0.2828 2.965 2.813 Ts1GC2a 0.2649 0.2806 3.173 2.534 Ts2GC2b 0.5257 0.5615 4.705 5.951 Ts2GC2c 0.5465 0.6898 4.284 5.862 Ts3GC2d 0.5363 0.4132 0.7954 0.7681 Ts3GC2e 0.2963 0.2718 3.289 3.684 Bs3GC1a 0.6812 0.9625 4.721 6.235 Bs3GC1b 0.5546 0.7118 4.369 5.928 Bs2GC1c 0.8525 1.239 4.496 6.227 Bs2GC1d 3.075 6.299 9.596 15.89 Bs1GC2a 2.77 31.37 9.917 217.1 Bs3GC2b 0.5595 1.068 6.207 11.18 Bs1GC3a 0.7436 0.7371 4.636 13.3 Bs3GC3b 1.279 2.957 17.81 123.8 Bs3GC4 9.257 90.78 97.5 738.9 Bs3GC5 1.305 4.177 6.465 10.25 MOR103 24.86 139.4 172.3 1490 Namilumab 177.2 798.3 1972 19820

Example 9: Immune Complexes of GM-CSF with Multispecific Antibodies Bind to Fcγ Receptors IIa and IIb

(32) To address the engagement of Fcγ receptors, immune complexes formed between GM-CSF and multispecific antibodies were tested for their capacity to bind to TZM-b1 cells expressing FcγRIIa and FcγRIIb receptors. Multispecific antibodies were mixed with 0.05 μg/ml of GM-CSF, or staining buffer. Samples were incubated at 37° C. for 30 min to allow the formation of immune complexes and then cooled down to 4° C. before adding them to TZM-b1 cells for 30 minutes. Cells were washed twice and stained with anti-human IgG Fcγ fragment specific F(ab′)2 fragment. Samples were analyzed on BD FACSCanto (BD Biosciences) and median intensity fluorescence was analyzed and compared between samples. Strong binding was observed with immune complexes formed by GM-CSF and different multispecific antibodies on FcγRIIa- and FcγRIIb-expressing cells (Table 11). In particular Ts3GC2d and Bs1GC3a showed the strongest binding to FcγRIIa and FcγRIIb in presence of GM-CSF, while they poorly bound the same FcRs in absence of GM-CSF (FIG. 7). Taken together, the above results suggest that GM-CSF in immune complexes with a multispecific monoclonal antibody can be cleared from the human body through an Fc-dependent mechanism.

(33) TABLE-US-00011 TABLE 11 Binding of immune complexes formed by GM-CSF and different multispecific mAbs to TZM-bl cells expressing FcγRIIa or FcγRIIb, as measured by flow cytometry using an anti-IgG Fc specific antibody. Shown is the median fluorescence intensity (MFI) of multispecific antibodies alone or complexed with low (1:10 mAb:GM-CSF ratio) or high GM-CSF concentrations (1:1 mAb:GM-CSF ratio), and the average ratio of binding in presence and in absence of GM-CSF. Dark grey cells indicate the GM-CSF complexes formed with multispecific antibodies that poorly bind to FcRs in absence of GM-CSF. FcgRIIa FcgRIIa FcgRIIa FcgRIIb FcgRIIb FcgRIIb (MFI) (MFI) (MFI) Average (MFI) (MFI) (MFI) Average w/o w/ low w/ high RATIO w/o w/ low w/ high RATIO Name GM-CSF GM-CSF GM-CSF FcgRIIa GM-CSF GM-CSF GM-CSF FcgRIIb Ts1GC1 52422 117643 215686 3.18 350077 527988 1230711 2.51 Ts1GC2a 2850 4131 9872 2.46 67215 135509 397219 3.96 Ts2GC2b 11341 135974 725556 37.98 168007 852217 1262147 6.29 Ts2GC2c 9989 91425 189657 14.07 379395 896907 712072 2.12 Ts3GC2d 1313 7898 122101 49.50 28963 343854 651881 17.19 Ts3GC2e 1869 2181 189634 51.31 52524 109361 1236720 12.81 Bs3GC1a 687 1270 65292 48.44 20590 175426 510410 16.65 Bs3GC1b 3548 2937 37183 5.65 53830 60053 378584 4.07 Bs2GC1c 1590 5003 427407 135.98 107869 416283 942605 6.30 Bs2GC1d 2906 10922 319374 56.83 125493 435244 1219560 6.59 Bs1GC2a 662 9141 566165 434.52 11787 398084 1437205 77.85 Bs3GC2b 6906 7127 268634 19.97 96743 181252 1303031 7.67 Bs1GC3a 694 14400 128441 102.91 18221 98121 752715 23.35 Bs3GC3b 650 8290 54962 48.66 2627 57837 432575 93.34 Bs3GC4 6733 24478 19481 3.26 122923 137194 62612 0.81 Bs3GC5 1105 30163 468183 225.50 176650 393019 1578476 5.58

(34) TABLE-US-00012 Table of Sequences and SEQ ID Numbers SEQ ID NO Description Sequence GCA7 ANTIBODY   1 CDRH1 aa gftvstny   2 CDRH2 aa lyaggvt   3 CDRH3 aa akhydsgystidhfds   4 CDRL1 aa qsvfytsknkny   5 CDRL2 aa was   6 CDRL2 long aa LIYwasTRE   7 CDRL3 aa qqyystpft   8 CDRH1 nuc varS1 GGATTCACCGTCAGTACCAACTAC   9 CDRH1 nuc varS2 GGGTTTACTGTGTCTACAAACTAC  10 CDRH1 nuc varN1 GGCTTTACTGTCTCTACAAACTAC  11 CDRH1 nuc varC1 GGCTTCACCGTGTCAACAAACTAC  12 CDRH1 nuc varC2 GGGTTTACCGTCTCTACAAACTAC  13 CDRH2 nuc varS1 CTTTATGCCGGAGGTGTCACA  14 CDRH2 nuc CTGTACGCTGGCGGGGTGACC varS2/N1/C2  15 CDRH2 nuc varC1 CTGTACGCCGGAGGCGTGACT  16 CDRH3 nuc varS1 GCGAAACACTATGATTCGGGATATTCTACCATAGAT CACTTTGACTCC  17 CDRH3 nuc varS2 GCCAAACACTATGATAGTGGGTACTCCACTATTGAC CATTTTGACTCT  18 CDRH3 nuc varN1 GCCAAACACTATGATAGTGGGTATAGCACAATCGAC CATTTTGACAGC  19 CDRH3 nuc varC1 GCAAAACACTACGATTCTGGGTATAGTACAATTGAC CATTTTGATTCT  20 CDRH3 nuc varC2 GCCAAACACTATGATAGTGGGTACAGTACCATTGAC CATTTCGATAGC  21 CDRL1 nuc varS1 CAGAGTGTTTTCTACACCTCCAAAAATAAAAACTAC  22 CDRL1 nuc varS2 CAGTCCGTCTTCTACACCAGTAAGAACAAAAACTAT  23 CDRL1 nuc varN1 CAGAGCGTGTTCTACACCAGTAAGAACAAAAACTAT  24 CDRL1 nuc varC1 CAGTCCGTGTTCTACACTTCTAAGAACAAAAACTAT  25 CDRL1 nuc varC2 CAGAGTGTCTTCTACACCAGTAAGAACAAAAACTAT  26 CDRL2 nuc varS1 TGGGCATCT  27 CDRL2 nuc TGGGCTAGC varS2/N1/C2  28 CDRL2 nuc varC1 TGGGCCTCA  29 CDRL2 long nuc CTCATTTACTGGGCATCTACCCGGGAG varS1  30 CDRL2 long nuc CTGATCTACTGGGCTAGCACTAGAGAG varS2/N1/C2  31 CDRL2 long nuc CTGATCTACTGGGCCTCAACCCGAGAG varC1  32 CDRL3 nuc varS1 CAGCAATATTATAGTACCCCTTTCACT  33 CDRL3 nuc varS2 CAGCAGTATTATTCTACCCCCTTCACA  34 CDRL3 nuc varN1 CAGCAGTATTACAGCACCCCATTCACA  35 CDRL3 nuc varC1 CAGCAGTACTATAGCACTCCATTCACC  36 CDRL3 nuc varC2 CAGCAGTATTATTCAACACCCTTCACA  37 heavy chain GVQLVQSGGGLVQPGGSLRLSCAASgftvstnyMSWVRQAP variable domain GKGLEWVSIlyaggvtRYADSVKTRFTISRDNSKNTLFLQM (VH) aa NALSAEDTAIYYCakhydsgystidhfdsWGQGTLVTVSS  38 light chain variable DIQMTQSPDSVAVSLGERATINCKSSqsvfytsknknyLAWFQ domain (VL) aa QKPGQPPKLLIYwasTRESGVPDRFSGSGSGTDFTLTISSL RPEDVAVYYCqqyystpftFGPGTKVDIK  39 heavy chain ggggtgcaactggtgcagtctgggggaggcttggtccagccgggggggtccctgaga variable domain ctctcctgtgcagcctctGGATTCACCGTCAGTACCAACTACatga (VH) nuc varS1 gctgggtccgccaggctccagggaaggggctggagtgggtctcaattCTTTAT GCCGGAGGTGTCACAaggtacgcagactccgtgaagaccagattcaccat ctccagagacaattccaagaacactctctttcttcaaatgaacgccctgagcgccgaggac acggctatatattactgtGCGAAACACTATGATTCGGGATATTC TACCATAGATCACTTTGACTCCtggggccagggaaccctggtcacc gtctcctca  40 heavy chain GGCGTGCAGCTGGTGCAGAGCGGCGGCGGCCTGGTG variable domain CAGCCTGGAGGGTCACTGAGACTGTCATGCGCAGCAA (VH) nuc varS2 GCGGGTTTACTGTGTCTACAAACTACATGTCTTGGG TGAGGCAGGCACCTGGAAAGGGACTGGAGTGGGTCTC AATCCTGTACGCTGGCGGGGTGACCCGGTATGCAGA CAGCGTCAAGACCCGGTTCACAATTAGCAGAGATAACT CCAAAAATACTCTGTTTCTGCAGATGAATGCCCTGTCC GCTGAAGACACCGCAATCTACTATTGCGCCAAACACTA TGATAGTGGGTACTCCACTATTGACCATTTTGACTC TTGGGGGCAGGGGACTCTGGTGACTGTCTCTTCA  41 heavy chain GGCGTCCAGCTGGTGCAGAGCGGAGGGGGCCTGGTG variable domain CAGCCTGGCGGGTCCCTGAGACTGAGTTGTGCCGCAA (VH) nuc varN1 GTGGCTTTACTGTCTCTACAAACTACATGTCTTGGG TGAGGCAGGCACCTGGAAAGGGACTGGAGTGGGTCTC AATCCTGTACGCTGGCGGGGTGACCCGGTATGCAGA CAGCGTCAAGACCCGGTTCACAATTAGCAGAGATAACT CCAAAAATACTCTGTTTCTGCAGATGAATGCCCTGTCC GCTGAAGACACCGCAATCTACTATTGCGCCAAACACTA TGATAGTGGGTATAGCACAATCGACCATTTTGACAG CTGGGGACAGGGAACTCTGGTGACAGTCTCATCA  42 heavy chain GGAGTGCAGCTGGTCCAGAGCGGAGGAGGACTGGTG variable domain CAGCCAGGAGGGTCACTGAGGCTGAGCTGCGCAGCTT (VH) nuc varC1 CCGGCTTCACCGTGTCAACAAACTACATGAGCTGGGT CCGCCAGGCACCTGGGAAGGGACTGGAGTGGGTGTCC ATCCTGTACGCCGGAGGCGTGACTCGATATGCTGAC TCTGTCAAGACTCGGTTCACCATCTCTAGAGATAACAG TAAGAACACCCTGTTTCTGCAGATGAATGCACTGAGT GCCGAAGACACAGCTATCTACTATTGTGCAAAACACTA CGATTCTGGGTATAGTACAATTGACCATTTTGATTC TTGGGGCCAGGGGACACTGGTGACTGTCAGCTCC  43 heavy chain GGCGTGCAGCTGGTCCAGAGCGGAGGCGGACTGGTCC variable domain AGCCCGGCGGATCACTGAGACTGTCATGTGCCGCAAG (VH) nuc varC2 CGGGTTTACCGTCTCTACAAACTACATGTCTTGGGT GAGGCAGGCACCTGGAAAGGGACTGGAGTGGGTCTCA ATCCTGTACGCTGGCGGGGTGACCCGGTATGCAGAC AGCGTCAAGACCCGGTTCACAATTAGCAGAGATAACTC CAAAAATACTCTGTTTCTGCAGATGAATGCCCTGTCCG CTGAAGACACCGCAATCTACTATTGCGCCAAACACTAT GATAGTGGGTACAGTACCATTGACCATTTCGATAGC TGGGGGCAGGGGACTCTGGTGACCGTCTCATCA  44 light chain variable gacatccagatgacccagtctccagactccgtggctgtgtctctgggcgagagggccacc domain (VL) nuc atcaactgcaagtccagcCAGAGTGTTTTCTACACCTCCAAAAA varS1 TAAAAACTACttagcttggttccagcagaaaccaggacagcctcctaaactgctc atttacTGGGCATCTacccgggagtccggggtccctgaccgattcagtggcag cgggtctgggacagatttcactctcaccatcagcagcctgcggcctgaagatgtggcagt ttattactgtCAGCAATATTATAGTACCCCTTTCACTttcggccc tgggaccaaagtggatatcaaa  45 light chain variable GACATTCAGATGACCCAGAGTCCTGACAGCGTGGCCG domain (VL) nuc TCTCACTGGGGGAAAGGGCTACTATCAATTGTAAAAG varS2 TTCACAGTCCGTCTTCTACACCAGTAAGAACAAAAAC TATCTGGCCTGGTTTCAGCAGAAGCCAGGCCAGCCCCC TAAACTGCTGATCTACTGGGCTAGCACTAGAGAGTCT GGAGTGCCAGACAGATTCTCTGGCAGTGGGTCAGGAA CCGACTTCACCCTGACAATTAGCTCCCTGAGGCCCGAA GACGTGGCCGTCTATTATTGTCAGCAGTATTATTCTA CCCCCTTCACATTCGGACCTGGGACTAAAGTGGATAT CAAA  46 light chain variable GACATTCAGATGACCCAGAGTCCTGATTCCGTGGCTG domain (VL) nuc TCTCACTGGGGGAGCGAGCAACTATTAACTGCAAGTC varN1 TTCACAGAGCGTGTTCTACACCAGTAAGAACAAAAAC TATCTGGCCTGGTTTCAGCAGAAGCCAGGCCAGCCCCC TAAACTGCTGATCTACTGGGCTAGCACTAGAGAGTCT GGAGTGCCAGACAGATTCTCTGGCAGTGGGTCAGGAA CCGACTTCACCCTGACAATTAGCTCCCTGAGGCCCGAA GACGTGGCCGTCTACTATTGTCAGCAGTATTACAGCA CCCCATTCACATTCGGCCCTGGAACCAAAGTGGATATT AAG  47 light chain variable GACATCCAGATGACTCAGTCTCCCGATAGTGTGGCCG domain (VL) nuc TCTCCCTGGGGGAGAGGGCTACAATTAACTGCAAGAG varC1 CTCCCAGTCCGTGTTCTACACTTCTAAGAACAAAAAC TATCTGGCATGGTTTCAGCAGAAGCCTGGACAGCCCC CTAAACTGCTGATCTACTGGGCCTCAACCCGAGAGAG CGGAGTCCCAGACAGATTCTCAGGCAGCGGGTCCGGA ACAGATTTTACCCTGACAATTTCTAGTCTGCGGCCTGA AGACGTGGCTGTCTACTATTGTCAGCAGTACTATAGC ACTCCATTCACCTTTGGCCCCGGGACAAAGGTGGATA TCAAA 48 light chain variable GATATTCAGATGACCCAGAGTCCTGATTCCGTCGCTG domain (VL) nuc TCTCACTGGGAGAAAGGGCAACCATTAACTGTAAAAG varC2 CTCACAGAGTGTCTTCTACACCAGTAAGAACAAAAAC TATCTGGCCTGGTTTCAGCAGAAGCCAGGCCAGCCCCC TAAACTGCTGATCTACTGGGCTAGCACTAGAGAGTCT GGAGTGCCAGACAGATTCTCTGGCAGTGGGTCAGGAA CCGACTTCACCCTGACAATTAGCTCCCTGAGGCCCGAA GACGTGGCCGTCTACTATTGTCAGCAGTATTATTCAA CACCCTTCACATTCGGACCAGGAACAAAAGTGGATAT TAAG GCA21 ANTIBODY  49 CDRH1 aa gftfsnyp  50 CDRH2 aa ilpdgnrk  51 CDRH3 aa trdgtyysnggvyqtyrrffdf  52 CDRL1 aa qnilnw  53 CDRL2 aa kas  54 CDRL2 long aa LIYkasDLQ  55 CDRL3 aa qhynsyplt  56 CDRH1 nuc GGATTCACCTTTTCGAACTATCCT  57 CDRH2 nuc ATTTTACCTGATGGGAACAGAAAA  58 CDRH3 nuc ACGAGAGATGGCACGTATTACTCTAATGGTGGTGT TTATCAGACATATCGAAGGTTCTTCGATTTC  59 CDRL1 nuc CAGAATATCCTTAATTGG  60 CDRL2 nuc AAGGCGTCT  61 CDRL2 long nuc ctgatatatAAGGCGTCTgatttacaa  62 CDRL3 nuc CAGCATTATAATAGTTATCCTCTCACT  63 heavy chain QVQLMESGGGVVQPGRSLRLSCSAFgftfsnypMHWVRQAP variable domain GKGLEWVAIilpdgnrkNYGRSVTGRFTISRDNSNNSLYLQ (VH) aa MNNLTTEDTAMYYCtrdgtyysnggvyqtyrrffdfWGRGTLVT VSS  64 light chain variable DIQMTQSPSTLSTSVGDRVTITCRASqnilnwLAWYQQKPG domain (VL) aa NAPNLLIYkasDLQSGVPSRFSGSGSGTEFTLTISSLQPDDF ATYYCqhynsypltFGGGTKVEIK  65 heavy chain caggtgcaattgatggagtctgggggaggcgtggtccagcctgggaggtccctgcgac variable domain tctcatgcagtgcctttGGATTCACCTTTTCGAACTATCCTatgca (VH) nuc ctgggtccgccaggctccaggcaagggacttgagtgggtggctatcATTTTACC TGATGGGAACAGAAAAaactatggaaggtccgtgacgggccgattcacc atctccagagacaattccaacaacagcctttatttgcaaatgaacaacctgacgactgagga cacggctatgtactattgtACGAGAGATGGCACGTATTACTCTA ATGGTGGTGTTTATCAGACATATCGAAGGTTCTTC GATTTCtggggccgtggcaccctggtcaccgtctcctca  66 light chain variable gacatccagatgacccagtctccttccaccctgtctacatctgtgggagacagagtcaccat domain (VL) nuc cacttgccgggccagtCAGAATATCCTTAATTGGttggcctggtatcaa cagaaaccagggaacgcccctaacctcctgatatatAAGGCGTCTgatttacaaa gtggggtcccctcaagattcagcggcagtgggtctgggacagaattcactctcaccatca gcagcctgcagcctgatgattttgcaacttattactgcCAGCATTATAATAG TTATCCTCTCACTttcggcggagggaccaaggtggaaatcaaa GCB59 ANTIBODY  67 CDRH1 aa GLSFSSSG  68 CDRH2 aa ISGSQNYK  69 CDRH3 aa VGGFPYWLPPSDFSGFHV  70 CDRL1 aa NIGSKS  71 CDRL2 aa ADN  72 CDRL2 long aa VVYADNDRP  73 CDRL3 aa QVWDGNTDHVV  74 CDRH1 nuc varS1 GGATTGTCCTTCAGTAGTTCAGGC  75 CDRH1 nuc GGCCTGTCCTTCAGCTCCTCTGGC varN1/N2  76 CDRH1 nuc varC1 GGGCTGAGCTTCAGCTCCTCTGGA  77 CDRH2 nuc varS1 ATTAGTGGTAGTCAGAACTACAAA  78 CDRH2 nuc ATTAGCGGGTCCCAGAATTACAAG varN1/N2  79 CDRH2 nuc varC1 ATTTCTGGCAGTCAGAATTACAAG  80 CDRH3 nuc varS1 GTGGGAGGTTTCCCCTATTGGTTACCCCCGAGCGAC TTCTCCGGTTTCCATGTC  81 CDRH3 nuc GTCGGCGGGTTTCCCTATTGGCTGCCTCCAAGCGAC varN1/N2 TTTTCAGGGTTTCATGTC  82 CDRH3 nuc varC1 GTCGGGGGATTTCCCTATTGGCTGCCCCCTTCCGAT TTCTCTGGCTTTCACGTG  83 CDRL1 nuc varS1 AACATTGGAAGTAAAAGT  84 CDRL1 nuc AACATCGGCAGCAAGAGC varN1/N2  85 CDRL1 nuc varC1 AACATCGGGTCTAAGAGT  86 CDRL2 nuc varS1 GCTGATAAC  87 CDRL2 nuc varN1/ GCTGACAAC N2  88 CDRL2 nuc varC1 GCCGACAAT  89 CDRL2 long nuc GTCGTCTATGCTGATAACGACAGGCCC varS1  90 CDRL2 long nuc GTGGTCTATGCTGACAACGATCGGCCC varN1/N2  91 CDRL2 long nuc GTGGTCTATGCCGACAATGATCGGCCA varC1  92 CDRL3 nuc var S1 CAGGTGTGGGATGGTAATACTGATCATGTGGTC  93 CDRL3 nuc CAGGTCTGGGATGGGAATACTGACCACGTCGTC varN1/N2  94 CDRL3 nuc varC1 CAGGTCTGGGACGGGAACACAGATCATGTGGTC  95 heavy chain EVQLVESGGDLVKAGGSLRLSCAVSGLSFSSSGMNWVRQ variable domain APGKGLEWISSISGSQNYKYYADSVKGRFVVSRDNARNFL (VH) aa YLQMDSLRAEDTAVYFCVGGFPYWLPPSDFSGFHVWG QGTTVTVSS  96 light chain variable SYVLTQPPSVSVAPGQTASLTCGGTNIGSKSVHWYQQKA domain (VL) aa GQAPVLVVYADNDRPSGVPERFSGSNSGNTATLTISRVEA EDESDYFCQVWDGNTDHVVFGGGTKLTVL  97 heavy chain gaggtacaattggtggagtctgggggagacctggtcaaggcgggggggtccctgaga variable domain ctctcctgtgccgtctctggattgtccttcagtagttcaggcatgaattgggtccgccag (VH) nuc varS1 gctccagggaaggggctggagtggatctcatcgattagtggtagtcagaactacaaa tactatgcagactcagtgaagggccgattcgtcgtctccagagacaacgcccgcaactttc tatatctgcaaatggacagcctgagggccgaggatacggctgtgtatttttgtgtggga ggtttcccctattggttacccccgagcgacttctccggtttccatgtctggggccaa gggaccacggtcaccgtctcctca  98 heavy chain GAGGTGCAGCTGGTGGAAAGCGGAGGGGATCTGGTG variable domain AAAGCAGGAGGGAGCCTGAGACTGTCATGCGCCGTGA (VH) nuc var N1 GCGGGCTGTCATTCAGCTCCTCTGGCATGAACTGGG TGCGACAGGCTCCTGGAAAGGGACTGGAGTGGATCAG TTCAATTAGCGGGTCCCAGAATTACAAGTACTATGCA GACTCTGTCAAAGGAAGGTTCGTGGTCAGCCGGGATA ACGCCAGAAATTTTCTGTATCTGCAGATGGACAGCCT GCGCGCCGAAGATACCGCCGTGTACTTCTGCGTCGGC GGGTTTCCCTATTGGCTGCCTCCAAGCGATTTCAGC GGATTTCATGTCTGGGGGCAGGGAACTACAGTGACC GTCTCATCA  99 heavy chain GAGGTGCAGCTGGTGGAAAGTGGGGGCGATCTGGTC variable domain AAAGCCGGAGGGTCTCTGCGACTGTCTTGTGCTGTGA (VH) nuc var N2 GCGGCCTGTCCTTCAGCTCCTCTGGCATGAACTGGG TGCGACAGGCTCCTGGAAAGGGACTGGAGTGGATCAG TTCAATTAGCGGGTCCCAGAATTACAAGTACTATGCA GACTCTGTCAAAGGAAGGTTCGTGGTCAGCCGGGATA ACGCCAGAAATTTTCTGTATCTGCAGATGGACAGCCT GCGCGCCGAAGATACCGCCGTGTACTTCTGCGTCGGC GGGTTTCCCTATTGGCTGCCTCCAAGCGACTTTTCA GGGTTTCATGTCTGGGGGCAGGGAACTACCGTGACC GTCTCATCT 100 heavy chain GAGGTGCAGCTGGTCGAATCTGGCGGGGACCTGGTG variable domain AAGGCAGGAGGCAGTCTGAGGCTGTCATGCGCCGTCT (VH) nuc varC1 CAGGGCTGAGCTTCAGCTCCTCTGGAATGAACTGGG TGCGCCAGGCACCAGGCAAAGGACTGGAGTGGATCAG TTCAATTTCTGGCAGTCAGAATTACAAGTACTATGCT GACAGTGTGAAAGGGCGATTCGTGGTCTCCCGGGATA ACGCAAGAAATTTTCTGTATCTGCAGATGGACAGCCT GAGAGCCGAAGATACTGCTGTGTACTTCTGTGTCGG GGGATTTCCCTATTGGCTGCCCCCTTCCGATTTCTC TGGCTTTCACGTGTGGGGACAGGGCACCACAGTGACC GTCAGCTCC 101 light chain variable TCATATGTGCTGACTCAACCACCCTCGGTGTCAGTGGC domain (VL) nuc CCCAGGACAGACGGCCAGTCTAACCTGTGGGGGAACT varS1 AACATTGGAAGTAAAAGTGTTCATTGGTACCAGCAAA AGGCAGGCCAGGCCCCTGTGTTGGTCGTCTATGCTGA TAACGACAGGCCCTCAGGGGTCCCTGAGCGATTCTCT GGCTCCAACTCTGGGAACACGGCCACCCTGACCATCAG CAGGGTCGAGGCCGAGGATGAGTCCGACTATTTCTGT CAGGTGTGGGATGGTAATACTGATCATGTGGTCTT CGGCGGAGGGACCAAGCTGACCGTCCTG 102 light chain variable TCTTACGTCCTGACCCAGCCACCTAGCGTGAGCGTCGC domain (VL) nuc ACCAGGGCAGACAGCTTCACTGACTTGCGGAGGCACA var N1 AACATTGGCAGCAAGAGCGTGCACTGGTACCAGCAGA AAGCCGGACAGGCTCCCGTCCTGGTGGTCTATGCTGA CAACGATCGGCCCTCTGGCGTGCCTGAAAGATTCAGC GGCTCCAACTCTGGGAATACCGCAACACTGACCATCAG TAGGGTCGAGGCCGAAGACGAGTCAGATTACTTTTGC CAGGTGTGGGACGGCAATACTGACCATGTCGTGTTC GGCGGCGGGACCAAACTGACTGTGCTG 103 light chain variable TCCTACGTCCTGACTCAGCCACCTAGCGTGTCCGTCGC domain (VL) nuc ACCTGGGCAGACAGCATCACTGACTTGCGGGGGAACC var N2 AACATCGGCAGCAAGAGCGTGCACTGGTACCAGCAGA AAGCCGGACAGGCTCCCGTCCTGGTGGTCTATGCTGA CAACGATCGGCCCTCTGGCGTGCCTGAAAGATTCAGC GGCTCCAACTCTGGGAATACCGCAACACTGACCATCAG TAGGGTCGAGGCCGAAGACGAGTCAGATTACTTTTGC CAGGTCTGGGATGGGAATACTGACCACGTCGTCTTC GGAGGCGGAACCAAACTGACTGTCCTG 104 light chain variable TCCTACGTGCTGACTCAGCCACCTAGCGTGTCCGTCGC domain (VL) nuc ACCTGGACAGACTGCCAGCCTGACCTGCGGAGGAACAA varC1 ACATCGGGTCTAAGAGTGTGCACTGGTACCAGCAGAA AGCCGGACAGGCTCCCGTCCTGGTGGTCTATGCCGAC AATGATCGGCCATCTGGCGTGCCCGAAAGATTCTCAG GAAGCAACTCCGGCAATACCGCTACACTGACTATTTCT AGGGTGGAGGCAGAAGACGAGAGTGATTATTTCTGT CAGGTCTGGGACGGGAACACAGATCATGTGGTCTTT GGAGGCGGGACCAAGCTGACAGTGCTG GCE536 ANTIBODY 105 CDRH1 aa GYVFTSYY 106 CDRH2 aa ISPGDVNT 107 CDRH3 aa ARGPRSKPPYLYFALDV 108 CDRL1 aa QSVSSSL 109 CDRL2 aa GAS 110 CDRL2 long aa LIYGASNRA 111 CDRL3 aa QHYGSRVT 112 CDRH1 nuc varS1 GGATACGTGTTCACCTCTTACTAT 113 CDRH1 nuc varS2 GGATACGTCTTTACCTCTTACTAT 114 CDRH1 nuc GGATACGTCTTCACCTCTTACTAT varN1/C1 115 CDRH2 nuc ATCTCTCCCGGAGACGTGAACACT varS1/S2/N1/C1 116 CDRH3 nuc varS1 GCTAGGGGGCCCCGCAGCAAGCCTCCTTATCTGTAT TTTGCTCTGGATGTG 117 CDRH3 nuc varS2 GCTAGGGGGCCCCGCAGCAAGCCTCCTTATCTGTAC TTCGCTCTGGATGTC 118 CDRH3 nuc GCTAGGGGGCCCCGCAGCAAGCCTCCTTATCTGTAT varN1/C1 TTCGCTCTGGATGTC 119 CDRL1 nuc varS1 CAGAGTGTCAGCAGCAGCCTC 120 CDRL1 nuc varS2 CAGTCTGTGAGCTCCTCTCTG 121 CDRL1 nuc CAGTCCGTGAGCTCCTCTCTG varN1/C1 122 CDRL2 nuc varS1 GGTGCATCC 123 CDRL2 nuc GGCGCCTCC varS2/N1/C1 124 CDRL2 long nuc CTCATCTACGGTGCATCCAATAGGGCC varS1 125 CDRL2 long nuc CTGATCTATGGCGCCTCCAACCGCGCT varS2/N1/C1 126 CDRL3 nuc varS1 CAGCACTATGGCTCACGGGTCACT 127 CDRL3 nuc varS2 CAGCACTATGGCAGCAGGGTCACT 128 CDRL3 nuc varN1 CAGCATTATGGGTCACGGGTCACT 129 CDRL3 nuc varC1 CAGCATTATGGAAGCAGGGTCACC 130 heavy chain QLQLVQSGTEVKKPGASVKVSCKSSGYVFTSYYLVWVRQ variable domain APGQGLEWMATISPGDVNTSYEQRFQGRVTVTTDASTN (VH) aa TVDMELRSLRSEDTAVYYCARGPRSKPPYLYFALDVWG QGTAVTVSS 131 light chain variable EIVLTQSPGTLSLSPGETAILSCRASQSVSSSLLAWYQQKP domain (VL) aa GQAPRLLIYGASNRATGIRGRFSGSGSGTDFTLTISRLEPE DFVLYYCQHYGSRVTFGQGTKLEIK 132 heavy chain CAGCTGCAGCTGGTCCAGTCAGGCACAGAGGTCAAAA variable domain AGCCAGGAGCATCAGTGAAGGTGTCTTGTAAGTCATC (VH) nuc varS1 AGGATACGTGTTCACCTCTTACTATCTGGTGTGGGT CCGGCAGGCACCAGGACAGGGACTGGAGTGGATGGCC ACAATCTCTCCCGGAGACGTGAACACTAGTTACGAAC AGCGATTCCAGGGCAGAGTGACCGTCACCACAGACGCT TCAACTAATACCGTGGATATGGAGCTGCGGAGCCTGA GATCCGAAGATACAGCCGTCTACTATTGCGCTAGGGG GCCCCGCAGCAAGCCTCCTTATCTGTATTTTGCTCT GGATGTGTGGGGGCAGGGGACCGCTGTCACCGTGTC AAGC 133 heavy chain CAGCTGCAGCTGGTCCAGTCAGGCACAGAAGTCAAAAA variable domain ACCCGGCGCAAGCGTGAAGGTCTCATGTAAATCATCA (VH) nuc varS2 GGATACGTCTTTACCTCTTACTATCTGGTGTGGGTC CGGCAGGCACCAGGACAGGGACTGGAGTGGATGGCCA CAATCTCTCCCGGAGACGTGAACACTAGTTACGAACA GCGATTCCAGGGCAGAGTGACCGTCACCACAGACGCT TCAACTAATACCGTGGATATGGAGCTGCGGAGCCTGA GATCCGAAGATACAGCCGTCTACTATTGCGCTAGGGG GCCCCGCAGCAAGCCTCCTTATCTGTACTTCGCTCTG GATGTCTGGGGGCAGGGGACCGCCGTCACCGTCTCAA GC 134 heavy chain CAGCTGCAGCTGGTCCAGAGCGGCACAGAGGTGAAAA variable domain AGCCAGGAGCATCAGTCAAAGTGTCTTGTAAGTCATC (VH) nuc varN1 AGGATACGTCTTCACCTCTTACTATCTGGTGTGGGT CCGGCAGGCACCAGGACAGGGACTGGAGTGGATGGCC ACAATCTCTCCCGGAGACGTGAACACTAGTTACGAAC AGCGATTCCAGGGCAGAGTGACCGTCACCACAGACGCT TCAACTAATACCGTGGATATGGAGCTGCGGAGCCTGA GATCCGAAGATACAGCCGTCTACTATTGCGCTAGGGG GCCCCGCAGCAAGCCTCCTTATCTGTATTTCGCTCT GGATGTCTGGGGGCAGGGAACAGCAGTCACCGTCTCT TCT 135 heavy chain CAGCTGCAGCTGGTCCAGAGCGGAACCGAAGTGAAGA variable domain AACCCGGCGCAAGCGTCAAAGTCTCATGCAAATCAAGC (VH) nuc varC1 GGATACGTCTTCACCTCTTACTATCTGGTGTGGGTC CGGCAGGCACCAGGACAGGGACTGGAGTGGATGGCCA CAATCTCTCCCGGAGACGTGAACACTAGTTACGAACA GCGATTCCAGGGCAGAGTGACCGTCACCACAGACGCT TCAACTAATACCGTGGATATGGAGCTGCGGAGCCTGA GATCCGAAGATACAGCCGTCTACTATTGCGCTAGGGG GCCCCGCAGCAAGCCTCCTTATCTGTATTTCGCTCT GGATGTCTGGGGGCAGGGAACAGCAGTCACCGTCTCA AGC 136 light chain variable gaaattgtgttgacgcagtctcctggcaccctgtctttgtctccaggggaaacagccatcct domain (VL) nuc ctcctgcagggccagtcagagtgtcagcagcagcctcttagcctggtaccagcaaaaa varS1 cctggccaggctcccaggctcctcatctacggtgcatccaatagggccactggcatcaga ggcaggtttagtggcagtgggtctgggacagacttcactctcaccatcagtagattggag cctgaagattttgtactttattactgtcagcactatggctcacgggtcacttttggccag gggaccaagctggagatcaaac 137 light chain variable GAAATCGTGCTGACCCAGTCTCCTGGAACTCTGTCTCT domain (VL) nuc GTCACCTGGCGAAACCGCAATCCTGTCCTGTAGGGCAA varS2 GTCAGTCTGTGAGCTCCTCTCTGCTGGCATGGTACC AGCAGAAGCCCGGACAGGCCCCTAGGCTGCTGATCTA TGGCGCCTCCAACCGCGCTACTGGCATTCGGGGGAGA TTCAGTGGCTCAGGGAGCGGAACCGACTTTACCCTGA CAATCAGCCGGCTGGAGCCCGAAGATTTCGTGCTGTA TTACTGTCAGCACTATGGCAGCAGGGTCACTTTTGG GCAGGGGACTAAACTGGAGATTAAA 138 light chain variable GAAATCGTCCTGACCCAGTCACCTGGCACCCTGAGTCT domain (VL) nuc GAGTCCTGGCGAAACAGCAATCCTGTCTTGTCGGGCT varN1 TCACAGTCCGTGAGCTCCTCTCTGCTGGCATGGTACC AGCAGAAGCCCGGACAGGCCCCTAGGCTGCTGATCTA TGGCGCCTCCAACCGCGCTACTGGCATTCGGGGGAGA TTCAGTGGCTCAGGGAGCGGAACCGACTTTACCCTGA CAATCAGCCGGCTGGAGCCCGAAGATTTCGTGCTGTA CTACTGTCAGCATTATGGGTCACGGGTCACTTTTGG GCAGGGGACTAAACTGGAAATCAAG 139 light chain variable GAGATTGTCCTGACCCAGTCACCTGGCACCCTGAGCCT domain (VL) nuc GAGTCCTGGAGAGACCGCTATTCTGTCTTGTCGGGCA varC1 TCACAGTCCGTGAGCTCCTCTCTGCTGGCATGGTACC AGCAGAAGCCCGGACAGGCCCCTAGGCTGCTGATCTA TGGCGCCTCCAACCGCGCTACTGGCATTCGGGGGAGA TTCAGTGGCTCAGGGAGCGGAACCGACTTTACCCTGA CAATCAGCCGGCTGGAGCCCGAAGATTTCGTGCTGTA CTATTGTCAGCATTATGGAAGCAGGGTCACCTTCGG ACAGGGAACTAAACTGGAAATCAAG Constant regions 140 IgG1 CH1-CH2- ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS CH3 aa WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQT YICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGG PSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNW YVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRE EMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTP PVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHN HYTQKSLSLSPGK 141 IgG CK aa RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQ WKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYE KHKVYACEVTHQGLSSPVTKSFNRGEC 142 IgG CL aa GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTV AWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQW KSHRSYSCQVTHEGSTVEKTVAPTECS 143 Short linker aa GGGGS 144 Long linker aa GGGGSGGGGSGGGGS 145 IgG1 CH1-CH2- gcgtcgaccaagggcccatcggtcttccccctggcaccctcctccaagagcacctctggg CH3 nucl varS1 ggcacagcggccctgggctgcctggtcaaggactacttccccgaacctgtgacggtctc gtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctacagtcctc aggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacccagac ctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagttgagcc caaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctgggggga ccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacccctga ggtcacatgcgtggtggtggacgtgagccacgaGgaTcctgaggtcaagttcaactg gtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggagcagta caacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctgaatgg caaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaaaccatc tccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatcccggga ggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatcccagcga catcgccgtggagtgggagagcaatgggcagccggagaacaactacaagaccacgcct cccgtgctggactccgacggctccttcttcctctatagcaagctcaccgtggacaagagca ggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcacaaccact acacgcagaagagcctctccctgtccccgggtaaa 146 IgG1 CH1-CH2 GCGTCGACCAAGGGCCCATCGGTCTTCCCCCTGGCACC CH3 nucl varS2 CTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGG GCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACG GTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGC ACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTAC TCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTT GGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAA TCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGC ACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCC CCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCC CTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGA GGATCCTGAAGTCAAGTTCAACTGGTACGTGGATGGC GTCGAGGTGCATAATGCCAAGACAAAACCCCGGGAGG AACAGTACAACTCAACTTATAGAGTCGTGAGCGTCCTG ACCGTGCTGCATCAGGACTGGCTGAACGGCAAAGAAT ACAAGTGCAAAGTGTCTAATAAGGCCCTGCCTGCTCCA ATCGAGAAAACAATTAGCAAGGCAAAAGGGCAGCCCAG GGAACCTCAGGTGTACACTCTGCCTCCAAGCCGCGAGG AAATGACCAAGAACCAGGTCTCCCTGACATGTCTGGTG AAAGGATTCTATCCTAGTGACATTGCCGTGGAGTGGG AATCAAATGGCCAGCCAGAGAACAATTACAAGACCACA CCCCCTGTGCTGGACTCTGATGGGAGTTTCTTTCTGT ATTCCAAGCTGACCGTGGATAAATCTAGATGGCAGCA GGGAAATGTCTTTAGCTGTTCCGTGATGCATGAGGCA CTGCACAACCATTACACCCAGAAATCACTGTCACTGTC CCCAGGAAAA 147 IgG CK nucl cgTacGgtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctg gaactgcctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtg gaaggtggataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggaca gcaaggacagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgag aaacacaaagtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaag agcttcaacaggggagagtgt 148 IgG CL nucl ggtcagcccaaggctgccccctcggtcactctgttcccgccctcctctgaggagcttcaag ccaacaaggccacactggtgtgtctcataagtgacttctacccgggagccgtgacagtgg cttggaaagcagatagcagccccgtcaaggcgggagtggagaccaccacaccctccaaa caaagcaacaacaagtacgcggccagcagctatctgagcctgacgcctgagcagtggaa gtcccacagaagctacagctgccaggtcacgcatgaagggagcaccgtggagaagaca gtggcccctacagaatgttca 149 Short linker nucl GGCGGGGGAGGCTCT 150 Long linker nucl GGCGGGGGAGGCTCTGGGGGAGGCGGGAGTGGAGGC GGGGGATCA Engineered chains of multtspecific antibodies 151 Ts1GC1 heavy QVQLMESGGGVVQPGRSLRLSCSAFGFTFSNYPMHWVR chain aa QAPGKGLEWVAIILPDGNRKNYGRSVTGRFTISRDNSNN SLYLQMNNLTTEDTAMYYCTRDGTYYSNGGVYQTYRR FFDFWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGVQLV QSGGGLVQPGGSLRLSCAASGFTVSTNYMSWVRQAPGK GLEWVSILYAGGVTRYADSVKTRFTISRDNSKNTLFLQM NALSAEDTAIYYCAKHYDSGYSTIDHFDSWGQGTLVTVS SGGGGSGGGGSGGGGSDIQMTQSPDSVAVSLGERATINC KSSQSVFYTSKNKNYLAWFQQKPGQPPKLLIYWASTRES GVPDRFSGSGSGTDFTLTISSLRPEDVAVYYCQQYYSTPF TFGPGTKVDIKGGGGSEVQLVESGGDLVKAGGSLRLSCA VSGLSFSSSGMNWVRQAPGKGLEWISSISGSQNYKYYADS VKGRFVVSRDNARNFLYLQMDSLRAEDTAVYFCVGGFP YWLPPSDFSGFHVWGQGTTVTVSSGGGGSGGGGSGGG GSSYVLTQPPSVSVAPGQTASLTCGGTNIGSKSVHWYQQ KAGQAPVLVVYADNDRPSGVPERFSGSNSGNTATLTISRV EAEDESDYFCQVWDGNTDHVVFGGGTKLTVL 152 Ts1GC1 heavy CAGGTGCAATTGATGGAGTCTGGGGGAGGCGTGGTC chain nucl CAGCCTGGGAGGTCCCTGCGACTCTCATGCAGTGCCT TTGGATTCACCTTTTCGAACTATCCTATGCACTGGGT CCGCCAGGCTCCAGGCAAGGGACTTGAGTGGGTGGCT ATCATTTTACCTGATGGGAACAGAAAAAACTATGGAA GGTCCGTGACGGGCCGATTCACCATCTCCAGAGACAAT TCCAACAACAGCCTTTATTTGCAAATGAACAACCTGAC GACTGAGGACACGGCTATGTACTATTGTACGAGAGAT GGCACGTATTACTCTAATGGTGGTGTTTATCAGACAT ATCGAAGGTTCTTCGATTTCTGGGGCCGTGGCACCCT GGTCACCGTCTCCTCAGCGTCGACCAAGGGCCCATCGG TCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGG GGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACT TCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCC CTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACA GTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCG TGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGC AACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAA GAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACAT GCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCG TCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGG TGGACGTGAGCCACGAGGATCCTGAAGTCAAGTTCAA CTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAG ACAAAACCCCGGGAGGAACAGTACAACTCAACTTATAG AGTCGTGAGCGTCCTGACCGTGCTGCATCAGGACTGG CTGAACGGCAAAGAATACAAGTGCAAAGTGTCTAATA AGGCCCTGCCTGCTCCAATCGAGAAAACAATTAGCAAG GCAAAAGGGCAGCCCAGGGAACCTCAGGTGTACACTC TGCCTCCAAGCCGCGAGGAAATGACCAAGAACCAGGTC TCCCTGACATGTCTGGTGAAAGGATTCTATCCTAGTG ACATTGCCGTGGAGTGGGAATCAAATGGCCAGCCAGA GAACAATTACAAGACCACACCCCCTGTGCTGGACTCTG ATGGGAGTTTCTTTCTGTATTCCAAGCTGACCGTGGA TAAATCTAGATGGCAGCAGGGAAATGTCTTTAGCTGT TCCGTGATGCATGAGGCACTGCACAACCATTACACCCA GAAATCACTGTCACTGTCCCCAGGAAAAGGCGGGGGA GGCTCTGGAGTGCAGCTGGTCCAGAGCGGAGGAGGAC TGGTGCAGCCAGGAGGGTCACTGAGGCTGAGCTGCGC AGCTTCCGGCTTCACCGTGTCAACAAACTACATGAGCT GGGTCCGCCAGGCACCTGGGAAGGGACTGGAGTGGG TGTCCATCCTGTACGCCGGAGGCGTGACTCGATATGC TGACTCTGTCAAGACTCGGTTCACCATCTCTAGAGATA ACAGTAAGAACACCCTGTTTCTGCAGATGAATGCACTG AGTGCCGAAGACACAGCTATCTACTATTGTGCAAAACA CTACGATTCTGGGTATAGTACAATTGACCATTTTGAT TCTTGGGGCCAGGGGACACTGGTGACTGTCAGCTCCG GCGGGGGAGGCTCTGGGGGAGGCGGGAGTGGAGGCG GGGGATCAGACATCCAGATGACTCAGTCTCCCGATAG TGTGGCCGTCTCCCTGGGGGAGAGGGCTACAATTAAC TGCAAGAGCTCCCAGTCCGTGTTCTACACTTCTAAGAA CAAAAACTATCTGGCATGGTTTCAGCAGAAGCCTGGA CAGCCCCCTAAACTGCTGATCTACTGGGCCTCAACCCG AGAGAGCGGAGTCCCAGACAGATTCTCAGGCAGCGGG TCCGGAACAGATTTTACCCTGACAATTTCTAGTCTGCG GCCTGAAGACGTGGCTGTCTACTATTGTCAGCAGTAC TATAGCACTCCATTCACCTTTGGCCCCGGGACAAAGGT GGATATCAAAGGCGGGGGAGGCTCTGAGGTGCAGCT GGTCGAATCTGGCGGGGACCTGGTGAAGGCAGGAGG CAGTCTGAGGCTGTCATGCGCCGTCTCAGGGCTGAGC TTCAGCTCCTCTGGAATGAACTGGGTGCGCCAGGCAC CAGGCAAAGGACTGGAGTGGATCAGTTCAATTTCTGG CAGTCAGAATTACAAGTACTATGCTGACAGTGTGAAA GGGCGATTCGTGGTCTCCCGGGATAACGCAAGAAATT TTCTGTATCTGCAGATGGACAGCCTGAGAGCCGAAGA TACTGCTGTGTACTTCTGTGTCGGGGGATTTCCCTAT TGGCTGCCCCCTTCCGATTTCTCTGGCTTTCACGTGT GGGGACAGGGCACCACAGTGACCGTCAGCTCCGGCGG GGGAGGCTCTGGGGGAGGCGGGAGTGGAGGCGGGG GATCATCCTACGTGCTGACTCAGCCACCTAGCGTGTCC GTCGCACCTGGACAGACTGCCAGCCTGACCTGCGGAG GAACAAACATCGGGTCTAAGAGTGTGCACTGGTACCA GCAGAAAGCCGGACAGGCTCCCGTCCTGGTGGTCTAT GCCGACAATGATCGGCCATCTGGCGTGCCCGAAAGAT TCTCAGGAAGCAACTCCGGCAATACCGCTACACTGACT ATTTCTAGGGTGGAGGCAGAAGACGAGAGTGATTATT TCTGTCAGGTCTGGGACGGGAACACAGATCATGTGGT CTTTGGAGGCGGGACCAAGCTGACAGTGCTG 153 Ts1GC2a heavy QLQLVQSGTEVKKPGASVKVSCKSSGYVFTSYYLVWVRQ chain aa APGQGLEWMATISPGDVNTSYEQRFQGRVTVTTDASTN TVDMELRSLRSEDTAVYYCARGPRSKPPYLYFALDVWGQ GTAVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP SSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGKGGGGSGVQLVQSGGGL VQPGGSLRLSCAASGFTVSTNYMSWVRQAPGKGLEWVSI LYAGGVTRYADSVKTRFTISRDNSKNTLFLQMNALSAED TAIYYCAKHYDSGYSTIDHFDSWGQGTLVTVSSGGGGSG GGGSGGGGSDIQMTQSPDSVAVSLGERATINCKSSQSVFY TSKNKNYLAWFQQKPGQPPKLLIYWASTRESGVPDRFSG SGSGTDFTLTISSLRPEDVAVYYCQQYYSTPFTFGPGTKV DIKGGGGSEVQLVESGGDLVKAGGSLRLSCAVSGLSFSSS GMNWVRQAPGKGLEWISSISGSQNYKYYADSVKGRFVVS RDNARNFLYLQMDSLRAEDTAVYFCVGGFPYWLPPSDF SGFHVWGQGTTVTVSSGGGGSGGGGSGGGGSSYVLTQP PSVSVAPGQTASLTCGGTNIGSKSVHWYQQKAGQAPVLV VYADNDRPSGVPERFSGSNSGNTATLTISRVEAEDESDYF CQVWDGNTDHVVFGGGTKLTVL 154 Ts1GC2a heavy CAGCTGCAGCTGGTCCAGTCAGGCACAGAGGTCAAAA AGCCAGGAGCATCAGTGAAGGTGTCTTGTAAGTCATC AGGATACGTGTTCACCTCTTACTATCTGGTGTGGGTC CGGCAGGCACCAGGACAGGGACTGGAGTGGATGGCCA CAATCTCTCCCGGAGACGTGAACACTAGTTACGAACAG CGATTCCAGGGCAGAGTGACCGTCACCACAGACGCTTC AACTAATACCGTGGATATGGAGCTGCGGAGCCTGAGA TCCGAAGATACAGCCGTCTACTATTGCGCTAGGGGGC CCCGCAGCAAGCCTCCTTATCTGTATTTTGCTCTGGAT GTGTGGGGGCAGGGGACCGCTGTCACCGTGTCAAGCG CGTCGACCAAGGGCCCATCGGTCTTCCCCCTGGCACCC TCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCCTGG GCTGCCTGGTCAAGGACTACTTCCCCGAACCTGTGACG GTCTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGC ACACCTTCCCGGCTGTCCTACAGTCCTCAGGACTCTAC TCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCAGCTT GGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAA TCTTGTGACAAAACTCACACATGCCCACCGTGCCCAGC ACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTCTTCC CCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCC CTGAGGTCACATGCGTGGTGGTGGACGTGAGCCACGA GGATCCTGAAGTCAAGTTCAACTGGTACGTGGATGGC GTCGAGGTGCATAATGCCAAGACAAAACCCCGGGAGG AACAGTACAACTCAACTTATAGAGTCGTGAGCGTCCTG ACCGTGCTGCATCAGGACTGGCTGAACGGCAAAGAAT ACAAGTGCAAAGTGTCTAATAAGGCCCTGCCTGCTCCA ATCGAGAAAACAATTAGCAAGGCAAAAGGGCAGCCCAG GGAACCTCAGGTGTACACTCTGCCTCCAAGCCGCGAGG AAATGACCAAGAACCAGGTCTCCCTGACATGTCTGGTG AAAGGATTCTATCCTAGTGACATTGCCGTGGAGTGGG AATCAAATGGCCAGCCAGAGAACAATTACAAGACCACA CCCCCTGTGCTGGACTCTGATGGGAGTTTCTTTCTGT ATTCCAAGCTGACCGTGGATAAATCTAGATGGCAGCA GGGAAATGTCTTTAGCTGTTCCGTGATGCATGAGGCA CTGCACAACCATTACACCCAGAAATCACTGTCACTGTC CCCAGGAAAAGGCGGGGGAGGCTCTGGAGTGCAGCTG GTCCAGAGCGGAGGAGGACTGGTGCAGCCAGGAGGG TCACTGAGGCTGAGCTGCGCAGCTTCCGGCTTCACCG TGTCAACAAACTACATGAGCTGGGTCCGCCAGGCACCT GGGAAGGGACTGGAGTGGGTGTCCATCCTGTACGCCG GAGGCGTGACTCGATATGCTGACTCTGTCAAGACTCG GTTCACCATCTCTAGAGATAACAGTAAGAACACCCTGT TTCTGCAGATGAATGCACTGAGTGCCGAAGACACAGC TATCTACTATTGTGCAAAACACTACGATTCTGGGTAT AGTACAATTGACCATTTTGATTCTTGGGGCCAGGGGA CACTGGTGACTGTCAGCTCCGGCGGGGGAGGCTCTGG GGGAGGCGGGAGTGGAGGCGGGGGATCAGACATCCA GATGACTCAGTCTCCCGATAGTGTGGCCGTCTCCCTG GGGGAGAGGGCTACAATTAACTGCAAGAGCTCCCAGT CCGTGTTCTACACTTCTAAGAACAAAAACTATCTGGCA TGGTTTCAGCAGAAGCCTGGACAGCCCCCTAAACTGCT GATCTACTGGGCCTCAACCCGAGAGAGCGGAGTCCCA GACAGATTCTCAGGCAGCGGGTCCGGAACAGATTTTA CCCTGACAATTTCTAGTCTGCGGCCTGAAGACGTGGC TGTCTACTATTGTCAGCAGTACTATAGCACTCCATTCA CCTTTGGCCCCGGGACAAAGGTGGATATCAAAGGCGG GGGAGGCTCTGAGGTGCAGCTGGTCGAATCTGGCGG GGACCTGGTGAAGGCAGGAGGCAGTCTGAGGCTGTCA TGCGCCGTCTCAGGGCTGAGCTTCAGCTCCTCTGGAA TGAACTGGGTGCGCCAGGCACCAGGCAAAGGACTGGA GTGGATCAGTTCAATTTCTGGCAGTCAGAATTACAAG TACTATGCTGACAGTGTGAAAGGGCGATTCGTGGTCT CCCGGGATAACGCAAGAAATTTTCTGTATCTGCAGAT GGACAGCCTGAGAGCCGAAGATACTGCTGTGTACTTC TGTGTCGGGGGATTTCCCTATTGGCTGCCCCCTTCCG ATTTCTCTGGCTTTCACGTGTGGGGACAGGGCACCAC AGTGACCGTCAGCTCCGGCGGGGGAGGCTCTGGGGG AGGCGGGAGTGGAGGCGGGGGATCATCCTACGTGCT GACTCAGCCACCTAGCGTGTCCGTCGCACCTGGACAGA CTGCCAGCCTGACCTGCGGAGGAACAAACATCGGGTC TAAGAGTGTGCACTGGTACCAGCAGAAAGCCGGACAG GCTCCCGTCCTGGTGGTCTATGCCGACAATGATCGGC CATCTGGCGTGCCCGAAAGATTCTCAGGAAGCAACTCC GGCAATACCGCTACACTGACTATTTCTAGGGTGGAGG CAGAAGACGAGAGTGATTATTTCTGTCAGGTCTGGGA CGGGAACACAGATCATGTGGTCTTTGGAGGCGGGACC AAGCTGACAGTGCTG 155 Ts2GC2b/Bs1GC2a EVQLVESGGDLVKAGGSLRLSCAVSGLSFSSSGMNWVRQ heavy chain aa APGKGLEWISSISGSQNYKYYADSVKGRFVVSRDNARNFL YLQMDSLRAEDTAVYFCVGGFPYWLPPSDFSGFHVWGQ GTTVTVSSGGGGSGGGGSGGGGSSYVLTQPPSVSVAPGQ TASLTCGGTNIGSKSVHWYQQKAGQAPVLVVYADNDRPS GVPERFSGSNSGNTATLTISRVEAEDESDYFCQVWDGNT DHVVFGGGTKLTVLGGGGSQLQLVQSGTEVKKPGASVK VSCKSSGYVFTSYYLVWVRQAPGQGLEWMATISPGDVN TSYEQRFQGRVTVTTDASTNTVDMELRSLRSEDTAVYYC ARGPRSKPPYLYFALDVWGQGTAVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK 156 Ts2GC2b/Bs1GC2a GAGGTGCAGCTGGTGGAAAGCGGAGGGGATCTGGTG heavy chain nucl AAAGCAGGAGGGAGCCTGAGACTGTCATGCGCCGTGA GCGGGCTGTCATTCAGCTCCTCTGGCATGAACTGGG TGCGACAGGCTCCTGGAAAGGGACTGGAGTGGATCAG TTCAATTAGCGGGTCCCAGAATTACAAGTACTATGCA GACTCTGTCAAAGGAAGGTTCGTGGTCAGCCGGGATA ACGCCAGAAATTTTCTGTATCTGCAGATGGACAGCCT GCGCGCCGAAGATACCGCCGTGTACTTCTGCGTCGGC GGGTTTCCCTATTGGCTGCCTCCAAGCGATTTCAGC GGATTTCATGTCTGGGGGCAGGGAACTACAGTGACC GTCTCATCAGGCGGGGGAGGCTCTGGGGGAGGCGGG AGTGGAGGCGGGGGATCATCTTACGTCCTGACCCAGC CACCTAGCGTGAGCGTCGCACCAGGGCAGACAGCTTCA CTGACTTGCGGAGGCACAAACATTGGCAGCAAGAGCG TGCACTGGTACCAGCAGAAAGCCGGACAGGCTCCCGTC CTGGTGGTCTATGCTGACAACGATCGGCCCTCTGGCG TGCCTGAAAGATTCAGCGGCTCCAACTCTGGGAATACC GCAACACTGACCATCAGTAGGGTCGAGGCCGAAGACG AGTCAGATTACTTTTGCCAGGTGTGGGACGGCAATA CTGACCATGTCGTGTTCGGCGGCGGGACCAAACTGAC TGTGCTGGGCGGGGGAGGCTCTCAGCTGCAGCTGGT CCAGTCAGGCACAGAAGTCAAAAAACCCGGCGCAAGCG TGAAGGTCTCATGTAAATCATCAGGATACGTCTTTAC CTCTTACTATCTGGTGTGGGTCCGGCAGGCACCAGGA CAGGGACTGGAGTGGATGGCCACAATCTCTCCCGGAG ACGTGAACACTAGTTACGAACAGCGATTCCAGGGCAG AGTGACCGTCACCACAGACGCTTCAACTAATACCGTGG ATATGGAGCTGCGGAGCCTGAGATCCGAAGATACAGC CGTCTACTATTGCGCTAGGGGGCCCCGCAGCAAGCCT CCTTATCTGTACTTCGCTCTGGATGTCTGGGGGCAG GGGACCGCCGTCACCGTCTCAAGCGCGTCGACCAAGG GCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGC ACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCA AGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAAC TCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCG TGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACC TACATCTGCAACGTGAATCACAAGCCCAGCAACACCAA GGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAA ACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCT GGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACA TGCGTGGTGGTGGACGTGAGCCACGAGGATCCTGAG GTCAAGTTCAACTGGTACGTGGACGGCGTGGAGGTGC ATAATGCCAAGACAAAGCCGCGGGAGGAGCAGTACAA CAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGC ACCAGGACTGGCTGAATGGCAAGGAGTACAAGTGCAA GGTCTCCAACAAAGCCCTCCCAGCCCCCATCGAGAAAA CCATCTCCAAAGCCAAAGGGCAGCCCCGAGAACCACAG GTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAA GAACCAGGTCAGCCTGACCTGCCTGGTCAAAGGCTTCT ATCCCAGCGACATCGCCGTGGAGTGGGAGAGCAATGG GCAGCCGGAGAACAACTACAAGACCACGCCTCCCGTGC TGGACTCCGACGGCTCCTTCTTCCTCTATAGCAAGCTC ACCGTGGACAAGAGCAGGTGGCAGCAGGGGAACGTCT TCTCATGCTCCGTGATGCATGAGGCTCTGCACAACCAC TACACGCAGAAGAGCCTCTCCCTGTCCCCGGGTAAA 157 Ts2GC2b/Bs2GC1c GVQLVQSGGGLVQPGGSLRLSCAASGFTVSTNYMSWVR light chain aa QAPGKGLEWVSILYAGGVTRYADSVKTRFTISRDNSKNT LFLQMNALSAEDTAIYYCAKHYDSGYSTIDHFDSWGQGT LVTVSSGGGGSGGGGSGGGGSDIQMTQSPDSVAVSLGER ATINCKSSQSVFYTSKNKNYLAWFQQKPGQPPKLLIYWAS TRESGVPDRFSGSGSGTDFTLTISSLRPEDVAVYYCQQYY STPFTFGPGTKVDIKGGGGSEIVLTQSPGTLSLSPGETAIL SCRASQSVSSSLLAWYQQKPGQAPRLLIYGASNRATGIRG RFSGSGSGTDFTLTISRLEPEDFVLYYCQHYGSRVTFGQG TKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPR EAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLS KADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 158 Ts2GC2b/Bs2GC1c GGCGTCCAGCTGGTGCAGAGCGGAGGGGGCCTGGTG light chain nucl CAGCCTGGCGGGTCCCTGAGACTGAGTTGTGCCGCAA GTGGCTTTACTGTCTCTACAAACTACATGTCTTGGG TGAGGCAGGCACCTGGAAAGGGACTGGAGTGGGTCTC AATCCTGTACGCTGGCGGGGTGACCCGGTATGCAGA CAGCGTCAAGACCCGGTTCACAATTAGCAGAGATAACT CCAAAAATACTCTGTTTCTGCAGATGAATGCCCTGTCC GCTGAAGACACCGCAATCTACTATTGCGCCAAACACTA TGATAGTGGGTATAGCACAATCGACCATTTTGACAG CTGGGGACAGGGAACTCTGGTGACAGTCTCATCAGGC GGGGGAGGCTCTGGGGGAGGCGGGAGTGGAGGCGG GGGATCAGACATTCAGATGACCCAGAGTCCTGATTCC GTGGCTGTCTCACTGGGGGAGCGAGCAACTATTAACT GCAAGTCTTCACAGAGCGTGTTCTACACCAGTAAGAA CAAAAACTATCTGGCCTGGTTTCAGCAGAAGCCAGGC CAGCCCCCTAAACTGCTGATCTACTGGGCTAGCACTA GAGAGTCTGGAGTGCCAGACAGATTCTCTGGCAGTGG GTCAGGAACCGACTTCACCCTGACAATTAGCTCCCTGA GGCCCGAAGACGTGGCCGTCTACTATTGTCAGCAGTA TTACAGCACCCCATTCACATTCGGCCCTGGAACCAAAG TGGATATTAAGGGCGGGGGAGGCTCTGAAATCGTGC TGACCCAGTCTCCTGGAACTCTGTCTCTGTCACCTGGC GAAACCGCAATCCTGTCCTGTAGGGCAAGTCAGTCTG TGAGCTCCTCTCTGCTGGCATGGTACCAGCAGAAGCC CGGACAGGCCCCTAGGCTGCTGATCTATGGCGCCTCC AACCGCGCTACTGGCATTCGGGGGAGATTCAGTGGCT CAGGGAGCGGAACCGACTTTACCCTGACAATCAGCCGG CTGGAGCCCGAAGATTTCGTGCTGTATTACTGTCAGC ACTATGGCAGCAGGGTCACTTTTGGGCAGGGGACTA AACTGGAGATTAAAcgTacGgtggctgcaccatctgtcttcatcttcccgcc atctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatc ccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccag gagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgac gctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagg gcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt 159 Ts2GC2c/Bs1GC3a EVQLVESGGDLVKAGGSLRLSCAVSGLSFSSSGMNWVRQ heavy chain aa APGKGLEWISSISGSQNYKYYADSVKGRFVVSRDNARNFL YLQMDSLRAEDTAVYFCVGGFPYWLPPSDFSGFHVWGQ GTTVTVSSGGGGSGGGGSGGGGSSYVLTQPPSVSVAPGQ TASLTCGGTNIGSKSVHWYQQKAGQAPVLVVYADNDRPS GVPERFSGSNSGNTATLTISRVEAEDESDYFCQVWDGNT DHVVFGGGTKLTVLGGGGSGVQLVQSGGGLVQPGGSLR LSCAASGFTVSTNYMSWVRQAPGKGLEWVSILYAGGVT RYADSVKTRFTISRDNSKNTLFLQMNALSAEDTAIYYCAK HYDSGYSTIDHFDSWGQGTLVTVSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK 160 Ts2GC2c/Bs1GC3a GAGGTGCAGCTGGTGGAAAGTGGGGGCGATCTGGTC heavy chain nucl AAAGCCGGAGGGTCTCTGCGACTGTCTTGTGCTGTGA GCGGCCTGTCCTTCAGCTCCTCTGGCATGAACTGGG TGCGACAGGCTCCTGGAAAGGGACTGGAGTGGATCAG TTCAATTAGCGGGTCCCAGAATTACAAGTACTATGCA GACTCTGTCAAAGGAAGGTTCGTGGTCAGCCGGGATA ACGCCAGAAATTTTCTGTATCTGCAGATGGACAGCCT GCGCGCCGAAGATACCGCCGTGTACTTCTGCGTCGGC GGGTTTCCCTATTGGCTGCCTCCAAGCGACTTTTCA GGGTTTCATGTCTGGGGGCAGGGAACTACCGTGACC GTCTCATCTGGCGGGGGAGGCTCTGGGGGAGGCGGG AGTGGAGGCGGGGGATCATCCTACGTCCTGACTCAGC CACCTAGCGTGTCCGTCGCACCTGGGCAGACAGCATCA CTGACTTGCGGGGGAACCAACATCGGCAGCAAGAGCG TGCACTGGTACCAGCAGAAAGCCGGACAGGCTCCCGTC CTGGTGGTCTATGCTGACAACGATCGGCCCTCTGGCG TGCCTGAAAGATTCAGCGGCTCCAACTCTGGGAATACC GCAACACTGACCATCAGTAGGGTCGAGGCCGAAGACG AGTCAGATTACTTTTGCCAGGTCTGGGATGGGAATA CTGACCACGTCGTCTTCGGAGGCGGAACCAAACTGAC TGTCCTGGGCGGGGGAGGCTCTGGCGTGCAGCTGGT GCAGAGCGGCGGCGGCCTGGTGCAGCCTGGAGGGTCA CTGAGACTGTCATGCGCAGCAAGCGGGTTTACTGTG TCTACAAACTACATGTCTTGGGTGAGGCAGGCACCTG GAAAGGGACTGGAGTGGGTCTCAATCCTGTACGCTG GCGGGGTGACCCGGTATGCAGACAGCGTCAAGACCCG GTTCACAATTAGCAGAGATAACTCCAAAAATACTCTGT TTCTGCAGATGAATGCCCTGTCCGCTGAAGACACCGCA ATCTACTATTGCGCCAAACACTATGATAGTGGGTACT CCACTATTGACCATTTTGACTCTTGGGGGCAGGGGA CTCTGGTGACTGTCTCTTCAGCGTCGACCAAGGGCCCA TCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGAC TACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAG GCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTC CTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGT GACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACA TCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTG GACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCA CACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGG GACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGT GGTGGTGGACGTGAGCCACGAGGATCCTGAGGTCAAG TTCAACTGGTACGTGGACGGCGTGGAGGTGCATAATG CCAAGACAAAGCCGCGGGAGGAGCAGTACAACAGCACG TACCGTGTGGTCAGCGTCCTCACCGTCCTGCACCAGGA CTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCC AACAAAGCCCTCCCAGCCCCCATCGAGAAAACCATCTCC AAAGCCAAAGGGCAGCCCCGAGAACCACAGGTGTACAC CCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAGG TCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGC GACATCGCCGTGGAGTGGGAGAGCAATGGGCAGCCGG AGAACAACTACAAGACCACGCCTCCCGTGCTGGACTCC GACGGCTCCTTCTTCCTCTATAGCAAGCTCACCGTGGA CAAGAGCAGGTGGCAGCAGGGGAACGTCTTCTCATGC TCCGTGATGCATGAGGCTCTGCACAACCACTACACGCA GAAGAGCCTCTCCCTGTCCCCGGGTAAA 161 Ts2GC2c/Bs2GC1d QLQLVQSGTEVKKPGASVKVSCKSSGYVFTSYYLVWVRQ light chain aa APGQGLEWMATISPGDVNTSYEQRFQGRVTVTTDASTN TVDMELRSLRSEDTAVYYCARGPRSKPPYLYFALDVWGQ GTAVTVSSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPG ETAILSCRASQSVSSSLLAWYQQKPGQAPRLLIYGASNRAT GIRGRFSGSGSGTDFTLTISRLEPEDFVLYYCQHYGSRVT FGQGTKLEIKGGGGSDIQMTQSPDSVAVSLGERATINCKS SQSVFYTSKNKNYLAWFQQKPGQPPKLLIYWASTRESGV PDRFSGSGSGTDFTLTISSLRPEDVAVYYCQQYYSTPFTF GPGTKVDIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNF YPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSST LTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC 162 Ts2GC2c/Bs2GC1d CAGCTGCAGCTGGTCCAGAGCGGCACAGAGGTGAAAA light chain nucl AGCCAGGAGCATCAGTCAAAGTGTCTTGTAAGTCATC AGGATACGTCTTCACCTCTTACTATCTGGTGTGGGT CCGGCAGGCACCAGGACAGGGACTGGAGTGGATGGCC ACAATCTCTCCCGGAGACGTGAACACTAGTTACGAAC AGCGATTCCAGGGCAGAGTGACCGTCACCACAGACGCT TCAACTAATACCGTGGATATGGAGCTGCGGAGCCTGA GATCCGAAGATACAGCCGTCTACTATTGCGCTAGGGG GCCCCGCAGCAAGCCTCCTTATCTGTATTTCGCTCT GGATGTCTGGGGGCAGGGAACAGCAGTCACCGTCTCT TCTGGCGGGGGAGGCTCTGGGGGAGGCGGGAGTGGA GGCGGGGGATCAGAAATCGTCCTGACCCAGTCACCTG GCACCCTGAGTCTGAGTCCTGGCGAAACAGCAATCCTG TCTTGTCGGGCTTCACAGTCCGTGAGCTCCTCTCTGC TGGCATGGTACCAGCAGAAGCCCGGACAGGCCCCTAG GCTGCTGATCTATGGCGCCTCCAACCGCGCTACTGGC ATTCGGGGGAGATTCAGTGGCTCAGGGAGCGGAACCG ACTTTACCCTGACAATCAGCCGGCTGGAGCCCGAAGAT TTCGTGCTGTACTACTGTCAGCATTATGGGTCACGG GTCACTTTTGGGCAGGGGACTAAACTGGAAATCAAGG GCGGGGGAGGCTCTGACATTCAGATGACCCAGAGTCC TGACAGCGTGGCCGTCTCACTGGGGGAAAGGGCTACT ATCAATTGTAAAAGTTCACAGTCCGTCTTCTACACCA GTAAGAACAAAAACTATCTGGCCTGGTTTCAGCAGAA GCCAGGCCAGCCCCCTAAACTGCTGATCTACTGGGCT AGCACTAGAGAGTCTGGAGTGCCAGACAGATTCTCTG GCAGTGGGTCAGGAACCGACTTCACCCTGACAATTAGC TCCCTGAGGCCCGAAGACGTGGCCGTCTATTATTGTC AGCAGTATTATTCTACCCCCTTCACATTCGGACCTGG GACTAAAGTGGATATCAAACGTACGGTGGCTGCACCA TCTGTCTTCATCTTCCCGCCATCTGATGAGCAGTTGAA ATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAAC TTCTATCCCAGAGAGGCCAAAGTACAGTGGAAGGTGG ATAACGCCCTCCAATCGGGTAACTCCCAGGAGAGTGTC ACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTCAG CAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAAC ACAAAGTCTACGCCTGCGAAGTCACCCATCAGGGCCTG AGCTCGCCCGTCACAAAGAGCTTCAACAGGGGAGAGT GT 163 Ts3GC2d heavy EVQLVESGGDLVKAGGSLRLSCAVSGLSFSSSGMNWVRQ chain aa APGKGLEWISSISGSQNYKYYADSVKGRFVVSRDNARNFL YLQMDSLRAEDTAVYFCVGGFPYWLPPSDFSGFHVWGQ GTTVTVSSGGGGSGGGGSGGGGSSYVLTQPPSVSVAPGQ TASLTCGGTNIGSKSVHWYQQKAGQAPVLVVYADNDRPS GVPERFSGSNSGNTATLTISRVEAEDESDYFCQVWDGNT DHVVFGGGTKLTVLGGGGSQLQLVQSGTEVKKPGASVK VSCKSSGYVFTSYYLVWVRQAPGQGLEWMATISPGDVN TSYEQRFQGRVTVTTDASTNTVDMELRSLRSEDTAVYYC ARGPRSKPPYLYFALDVWGQGTAVTVSSASTKGPSVFPL APSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSN TKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPK DTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHN AKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVS NKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVS LTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGS FFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGKGGGGSGVQLVQSGGGLVQPGGSLRLSCAASGFTV STNYMSWVRQAPGKGLEWVSILYAGGVTRYADSVKTRF TISRDNSKNTLFLQMNALSAEDTAIYYCAKHYDSGYSTID HFDSWGQGTLVTVSSGGGGSGGGGSGGGGSDIQMTQSP DSVAVSLGERATINCKSSQSVFYTSKNKNYLAWFQQKPG QPPKLLIYWASTRESGVPDRFSGSGSGTDFTLTISSLRPED VAVYYCQQYYSTPFTFGPGTKVDIK 164 Ts3GC2d heavy GAGGTGCAGCTGGTGGAAAGCGGAGGGGATCTGGTG chain nucl AAAGCAGGAGGGAGCCTGAGACTGTCATGCGCCGTGA GCGGGCTGTCATTCAGCTCCTCTGGCATGAACTGGG TGCGACAGGCTCCTGGAAAGGGACTGGAGTGGATCAG TTCAATTAGCGGGTCCCAGAATTACAAGTACTATGCA GACTCTGTCAAAGGAAGGTTCGTGGTCAGCCGGGATA ACGCCAGAAATTTTCTGTATCTGCAGATGGACAGCCT GCGCGCCGAAGATACCGCCGTGTACTTCTGCGTCGGC GGGTTTCCCTATTGGCTGCCTCCAAGCGATTTCAGC GGATTTCATGTCTGGGGGCAGGGAACTACAGTGACC GTCTCATCAGGCGGGGGAGGCTCTGGGGGAGGCGGG AGTGGAGGCGGGGGATCATCTTACGTCCTGACCCAGC CACCTAGCGTGAGCGTCGCACCAGGGCAGACAGCTTCA CTGACTTGCGGAGGCACAAACATTGGCAGCAAGAGCG TGCACTGGTACCAGCAGAAAGCCGGACAGGCTCCCGTC CTGGTGGTCTATGCTGACAACGATCGGCCCTCTGGCG TGCCTGAAAGATTCAGCGGCTCCAACTCTGGGAATACC GCAACACTGACCATCAGTAGGGTCGAGGCCGAAGACG AGTCAGATTACTTTTGCCAGGTGTGGGACGGCAATA CTGACCATGTCGTGTTCGGCGGCGGGACCAAACTGAC TGTGCTGGGCGGGGGAGGCTCTCAGCTGCAGCTGGT CCAGTCAGGCACAGAAGTCAAAAAACCCGGCGCAAGCG TGAAGGTCTCATGTAAATCATCAGGATACGTCTTTAC CTCTTACTATCTGGTGTGGGTCCGGCAGGCACCAGGA CAGGGACTGGAGTGGATGGCCACAATCTCTCCCGGAG ACGTGAACACTAGTTACGAACAGCGATTCCAGGGCAG AGTGACCGTCACCACAGACGCTTCAACTAATACCGTGG ATATGGAGCTGCGGAGCCTGAGATCCGAAGATACAGC CGTCTACTATTGCGCTAGGGGGCCCCGCAGCAAGCCT CCTTATCTGTACTTCGCTCTGGATGTCTGGGGGCAG GGGACCGCCGTCACCGTCTCAAGCGCGTCGACCAAGG GCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGC ACCTCTGGGGGCACAGCGGCCCTGGGCTGCCTGGTCA AGGACTACTTCCCCGAACCTGTGACGGTCTCGTGGAAC TCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCCGGC TGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCG TGGTGACCGTGCCCTCCAGCAGCTTGGGCACCCAGACC TACATCTGCAACGTGAATCACAAGCCCAGCAACACCAA GGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAA ACTCACACATGCCCACCGTGCCCAGCACCTGAACTCCT GGGGGGACCGTCAGTCTTCCTCTTCCCCCCAAAACCCA AGGACACCCTCATGATCTCCCGGACCCCTGAGGTCACA TGCGTGGTGGTGGACGTGAGCCACGAGGATCCTGAAG TCAAGTTCAACTGGTACGTGGATGGCGTCGAGGTGCA TAATGCCAAGACAAAACCCCGGGAGGAACAGTACAACT CAACTTATAGAGTCGTGAGCGTCCTGACCGTGCTGCA TCAGGACTGGCTGAACGGCAAAGAATACAAGTGCAAA GTGTCTAATAAGGCCCTGCCTGCTCCAATCGAGAAAAC AATTAGCAAGGCAAAAGGGCAGCCCAGGGAACCTCAG GTGTACACTCTGCCTCCAAGCCGCGAGGAAATGACCAA GAACCAGGTCTCCCTGACATGTCTGGTGAAAGGATTC TATCCTAGTGACATTGCCGTGGAGTGGGAATCAAATG GCCAGCCAGAGAACAATTACAAGACCACACCCCCTGTG CTGGACTCTGATGGGAGTTTCTTTCTGTATTCCAAGC TGACCGTGGATAAATCTAGATGGCAGCAGGGAAATGT CTTTAGCTGTTCCGTGATGCATGAGGCACTGCACAAC CATTACACCCAGAAATCACTGTCACTGTCCCCAGGAAA AGGCGGGGGAGGCTCTGGCGTGCAGCTGGTCCAGAG CGGAGGCGGACTGGTCCAGCCCGGCGGATCACTGAGA CTGTCATGTGCCGCAAGCGGGTTTACCGTCTCTACAA ACTACATGTCTTGGGTGAGGCAGGCACCTGGAAAGGG ACTGGAGTGGGTCTCAATCCTGTACGCTGGCGGGGT GACCCGGTATGCAGACAGCGTCAAGACCCGGTTCACAA TTAGCAGAGATAACTCCAAAAATACTCTGTTTCTGCAG ATGAATGCCCTGTCCGCTGAAGACACCGCAATCTACTA TTGCGCCAAACACTATGATAGTGGGTACAGTACCAT TGACCATTTCGATAGCTGGGGGCAGGGGACTCTGGT GACCGTCTCATCAGGCGGGGGAGGCTCTGGGGGAGG CGGGAGTGGAGGCGGGGGATCAGATATTCAGATGACC CAGAGTCCTGATTCCGTCGCTGTCTCACTGGGAGAAA GGGCAACCATTAACTGTAAAAGCTCACAGAGTGTCTT CTACACCAGTAAGAACAAAAACTATCTGGCCTGGTTT CAGCAGAAGCCAGGCCAGCCCCCTAAACTGCTGATCTA CTGGGCTAGCACTAGAGAGTCTGGAGTGCCAGACAGA TTCTCTGGCAGTGGGTCAGGAACCGACTTCACCCTGA CAATTAGCTCCCTGAGGCCCGAAGACGTGGCCGTCTAC TATTGTCAGCAGTATTATTCAACACCCTTCACATTCG GACCAGGAACAAAAGTGGATATTAAG 165 Ts3GC2e heavy EVQLVESGGDLVKAGGSLRLSCAVSGLSFSSSGMNWVRQ chain aa APGKGLEWISSISGSQNYKYYADSVKGRFVVSRDNARNFL YLQMDSLRAEDTAVYFCVGGFPYWLPPSDFSGFHVWGQ GTTVTVSSGGGGSGGGGSGGGGSSYVLTQPPSVSVAPGQ TASLTCGGTNIGSKSVHWYQQKAGQAPVLVVYADNDRPS GVPERFSGSNSGNTATLTISRVEAEDESDYFCQVWDGNT DHVVFGGGTKLTVLGGGGSGVQLVQSGGGLVQPGGSLR LSCAASGFTVSTNYMSWVRQAPGKGLEWVSILYAGGVT RYADSVKTRFTISRDNSKNTLFLQMNALSAEDTAIYYCAK HYDSGYSTIDHFDSWGQGTLVTVSSASTKGPSVFPLAPSS KSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFP AVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLM ISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKP REEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVK GFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKL TVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGKG GGGSQLQLVQSGTEVKKPGASVKVSCKSSGYVFTSYYLV WVRQAPGQGLEWMATISPGDVNTSYEQRFQGRVTVTT DASTNTVDMELRSLRSEDTAVYYCARGPRSKPPYLYFAL DVWGQGTAVTVSSGGGGSGGGGSGGGGSEIVLTQSPGT LSLSPGETAILSCRASQSVSSSLLAWYQQKPGQAPRLLIYG ASNRATGIRGRFSGSGSGTDFTLTISRLEPEDFVLYYCQH YGSRVTFGQGTKLEIK 166 Ts3GC2e heavy GAGGTGCAGCTGGTGGAAAGTGGGGGCGATCTGGTC chain nucl AAAGCCGGAGGGTCTCTGCGACTGTCTTGTGCTGTGA GCGGCCTGTCCTTCAGCTCCTCTGGCATGAACTGGG TGCGACAGGCTCCTGGAAAGGGACTGGAGTGGATCAG TTCAATTAGCGGGTCCCAGAATTACAAGTACTATGCA GACTCTGTCAAAGGAAGGTTCGTGGTCAGCCGGGATA ACGCCAGAAATTTTCTGTATCTGCAGATGGACAGCCT GCGCGCCGAAGATACCGCCGTGTACTTCTGCGTCGGC GGGTTTCCCTATTGGCTGCCTCCAAGCGACTTTTCA GGGTTTCATGTCTGGGGGCAGGGAACTACCGTGACC GTCTCATCTGGCGGGGGAGGCTCTGGGGGAGGCGGG AGTGGAGGCGGGGGATCATCCTACGTCCTGACTCAGC CACCTAGCGTGTCCGTCGCACCTGGGCAGACAGCATCA CTGACTTGCGGGGGAACCAACATCGGCAGCAAGAGCG TGCACTGGTACCAGCAGAAAGCCGGACAGGCTCCCGTC CTGGTGGTCTATGCTGACAACGATCGGCCCTCTGGCG TGCCTGAAAGATTCAGCGGCTCCAACTCTGGGAATACC GCAACACTGACCATCAGTAGGGTCGAGGCCGAAGACG AGTCAGATTACTTTTGCCAGGTCTGGGATGGGAATA CTGACCACGTCGTCTTCGGAGGCGGAACCAAACTGAC TGTCCTGGGCGGGGGAGGCTCTGGCGTGCAGCTGGT GCAGAGCGGCGGCGGCCTGGTGCAGCCTGGAGGGTCA CTGAGACTGTCATGCGCAGCAAGCGGGTTTACTGTG TCTACAAACTACATGTCTTGGGTGAGGCAGGCACCTG GAAAGGGACTGGAGTGGGTCTCAATCCTGTACGCTG GCGGGGTGACCCGGTATGCAGACAGCGTCAAGACCCG GTTCACAATTAGCAGAGATAACTCCAAAAATACTCTGT TTCTGCAGATGAATGCCCTGTCCGCTGAAGACACCGCA ATCTACTATTGCGCCAAACACTATGATAGTGGGTACT CCACTATTGACCATTTTGACTCTTGGGGGCAGGGGA CTCTGGTGACTGTCTCTTCAGCGTCGACCAAGGGCCCA TCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGAC TACTTCCCCGAACCTGTGACGGTCTCGTGGAACTCAG GCGCCCTGACCAGCGGCGTGCACACCTTCCCGGCTGTC CTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGT GACCGTGCCCTCCAGCAGCTTGGGCACCCAGACCTACA TCTGCAACGTGAATCACAAGCCCAGCAACACCAAGGTG GACAAGAGAGTTGAGCCCAAATCTTGTGACAAAACTCA CACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGG GACCGTCAGTCTTCCTCTTCCCCCCAAAACCCAAGGAC ACCCTCATGATCTCCCGGACCCCTGAGGTCACATGCGT GGTGGTGGACGTGAGCCACGAGGATCCTGAAGTCAAG TTCAACTGGTACGTGGATGGCGTCGAGGTGCATAATG CCAAGACAAAACCCCGGGAGGAACAGTACAACTCAACT TATAGAGTCGTGAGCGTCCTGACCGTGCTGCATCAGG ACTGGCTGAACGGCAAAGAATACAAGTGCAAAGTGTC TAATAAGGCCCTGCCTGCTCCAATCGAGAAAACAATTA GCAAGGCAAAAGGGCAGCCCAGGGAACCTCAGGTGTA CACTCTGCCTCCAAGCCGCGAGGAAATGACCAAGAACC AGGTCTCCCTGACATGTCTGGTGAAAGGATTCTATCC TAGTGACATTGCCGTGGAGTGGGAATCAAATGGCCAG CCAGAGAACAATTACAAGACCACACCCCCTGTGCTGGA CTCTGATGGGAGTTTCTTTCTGTATTCCAAGCTGACC GTGGATAAATCTAGATGGCAGCAGGGAAATGTCTTTA GCTGTTCCGTGATGCATGAGGCACTGCACAACCATTAC ACCCAGAAATCACTGTCACTGTCCCCAGGAAAAGGCGG GGGAGGCTCTCAGCTGCAGCTGGTCCAGAGCGGAACC GAAGTGAAGAAACCCGGCGCAAGCGTCAAAGTCTCAT GCAAATCAAGCGGATACGTCTTCACCTCTTACTATCT GGTGTGGGTCCGGCAGGCACCAGGACAGGGACTGGA GTGGATGGCCACAATCTCTCCCGGAGACGTGAACACT AGTTACGAACAGCGATTCCAGGGCAGAGTGACCGTCA CCACAGACGCTTCAACTAATACCGTGGATATGGAGCTG CGGAGCCTGAGATCCGAAGATACAGCCGTCTACTATT GCGCTAGGGGGCCCCGCAGCAAGCCTCCTTATCTGT ATTTCGCTCTGGATGTCTGGGGGCAGGGAACAGCAG TCACCGTCTCAAGCGGCGGGGGAGGCTCTGGGGGAG GCGGGAGTGGAGGCGGGGGATCAGAGATTGTCCTGA CCCAGTCACCTGGCACCCTGAGCCTGAGTCCTGGAGAG ACCGCTATTCTGTCTTGTCGGGCATCACAGTCCGTGA GCTCCTCTCTGCTGGCATGGTACCAGCAGAAGCCCGG ACAGGCCCCTAGGCTGCTGATCTATGGCGCCTCCAAC CGCGCTACTGGCATTCGGGGGAGATTCAGTGGCTCAG GGAGCGGAACCGACTTTACCCTGACAATCAGCCGGCT GGAGCCCGAAGATTTCGTGCTGTACTATTGTCAGCAT TATGGAAGCAGGGTCACCTTCGGACAGGGAACTAAAC TGGAAATCAAG 167 Bs3GC1a heavy QLQLVQSGTEVKKPGASVKVSCKSSGYVFTSYYLVWVRQ chain aa APGQGLEWMATISPGDVNTSYEQRFQGRVTVTTDASTN TVDMELRSLRSEDTAVYYCARGPRSKPPYLYFALDVWGQ GTAVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP SSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGKGGGGSGVQLVQSGGGL VQPGGSLRLSCAASGFTVSTNYMSWVRQAPGKGLEWVSI LYAGGVTRYADSVKTRFTISRDNSKNTLFLQMNALSAED TAIYYCAKHYDSGYSTIDHFDSWGQGTLVTVSSGGGGSG GGGSGGGGSDIQMTQSPDSVAVSLGERATINCKSSQSVFY TSKNKNYLAWFQQKPGQPPKLLIYWASTRESGVPDRFSG SGSGTDFTLTISSLRPEDVAVYYCQQYYSTPFTFGPGTKV DIK 168 Bs3GC1a heavy CAGCTGCAGCTGGTCCAGTCAGGCACAGAGGTCAAAA chain nucl AGCCAGGAGCATCAGTGAAGGTGTCTTGTAAGTCATC AGGATACGTGTTCACCTCTTACTATCTGGTGTGGGT CCGGCAGGCACCAGGACAGGGACTGGAGTGGATGGCC ACAATCTCTCCCGGAGACGTGAACACTAGTTACGAAC AGCGATTCCAGGGCAGAGTGACCGTCACCACAGACGCT TCAACTAATACCGTGGATATGGAGCTGCGGAGCCTGA GATCCGAAGATACAGCCGTCTACTATTGCGCTAGGGG GCCCCGCAGCAAGCCTCCTTATCTGTATTTTGCTCT GGATGTGTGGGGGCAGGGGACCGCTGTCACCGTGTC AAGCGCGTCGACCAAGGGCCCATCGGTCTTCCCCCTGG CACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCC CTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGT GACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGC GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACT CTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCA GCTTGGGCACCCAGACCTACATCTGCAACGTGAATCAC AAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCC CAAATCTTGTGACAAAACTCACACATGCCCACCGTGCC CAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTC TTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGG ACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCC ACGAGGATCCTGAAGTCAAGTTCAACTGGTACGTGGA TGGCGTCGAGGTGCATAATGCCAAGACAAAACCCCGG GAGGAACAGTACAACTCAACTTATAGAGTCGTGAGCG TCCTGACCGTGCTGCATCAGGACTGGCTGAACGGCAA AGAATACAAGTGCAAAGTGTCTAATAAGGCCCTGCCT GCTCCAATCGAGAAAACAATTAGCAAGGCAAAAGGGCA GCCCAGGGAACCTCAGGTGTACACTCTGCCTCCAAGCC GCGAGGAAATGACCAAGAACCAGGTCTCCCTGACATG TCTGGTGAAAGGATTCTATCCTAGTGACATTGCCGTG GAGTGGGAATCAAATGGCCAGCCAGAGAACAATTACA AGACCACACCCCCTGTGCTGGACTCTGATGGGAGTTT CTTTCTGTATTCCAAGCTGACCGTGGATAAATCTAGA TGGCAGCAGGGAAATGTCTTTAGCTGTTCCGTGATGC ATGAGGCACTGCACAACCATTACACCCAGAAATCACTG TCACTGTCCCCAGGAAAAGGCGGGGGAGGCTCTGGCG TGCAGCTGGTCCAGAGCGGAGGCGGACTGGTCCAGCC CGGCGGATCACTGAGACTGTCATGTGCCGCAAGCGGG TTTACCGTCTCTACAAACTACATGTCTTGGGTGAGGC AGGCACCTGGAAAGGGACTGGAGTGGGTCTCAATCCT GTACGCTGGCGGGGTGACCCGGTATGCAGACAGCGT CAAGACCCGGTTCACAATTAGCAGAGATAACTCCAAAA ATACTCTGTTTCTGCAGATGAATGCCCTGTCCGCTGA AGACACCGCAATCTACTATTGCGCCAAACACTATGATA GTGGGTACAGTACCATTGACCATTTCGATAGCTGGG GGCAGGGGACTCTGGTGACCGTCTCATCAGGCGGGGG AGGCTCTGGGGGAGGCGGGAGTGGAGGCGGGGGATC AGATATTCAGATGACCCAGAGTCCTGATTCCGTCGCT GTCTCACTGGGAGAAAGGGCAACCATTAACTGTAAAA GCTCACAGAGTGTCTTCTACACCAGTAAGAACAAAAA CTATCTGGCCTGGTTTCAGCAGAAGCCAGGCCAGCCCC CTAAACTGCTGATCTACTGGGCTAGCACTAGAGAGTC TGGAGTGCCAGACAGATTCTCTGGCAGTGGGTCAGGA ACCGACTTCACCCTGACAATTAGCTCCCTGAGGCCCGA AGACGTGGCCGTCTACTATTGTCAGCAGTATTATTCA ACACCCTTCACATTCGGACCAGGAACAAAAGTGGATA TTAAG 169 Bs3GC1b heavy GVQLVQSGGGLVQPGGSLRLSCAASGFTVSTNYMSWVR chain aa QAPGKGLEWVSILYAGGVTRYADSVKTRFTISRDNSKNT LFLQMNALSAEDTAIYYCAKHYDSGYSTIDHFDSWGQGT LVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPE PVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSL GTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPCPAP ELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPE VKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLH QDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVY TLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPE NNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGKGGGGSQLQLVQSGTEVKKP GASVKVSCKSSGYVFTSYYLVWVRQAPGQGLEWMATISP GDVNTSYEQRFQGRVTVTTDASTNTVDMELRSLRSEDT AVYYCARGPRSKPPYLYFALDVWGQGTAVTVSSGGGGS GGGGSGGGGSEIVLTQSPGTLSLSPGETAILSCRASQSVSS SLLAWYQQKPGQAPRLLIYGASNRATGIRGRFSGSGSGT DFTLTISRLEPEDFVLYYCQHYGSRVTFGQGTKLEIK 170 Bs3GC1b heavy ggggtgcaactggtgcagtctgggggaggcttggtccagccgggggggtccctgaga chain nucl ctctcctgtgcagcctctGGATTCACCGTCAGTACCAACTACatga gctgggtccgccaggctccagggaaggggctggagtgggtctcaattCTTTAT GCCGGAGGTGTCACAaggtacgcagactccgtgaagaccagattcaccat ctccagagacaattccaagaacactctctttcttcaaatgaacgccctgagcgccgaggac acggctatatattactgtGCGAAACACTATGATTCGGGATATTC TACCATAGATCACTTTGACTCCtggggccagggaaccctggtcacc gtctcctcaGCGTCGACCAAGGGCCCATCGGTCTTCCCCCTG GCACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGC CCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTG TGACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGG CGTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGAC TCTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGC AGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCA CAAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGC CCAAATCTTGTGACAAAACTCACACATGCCCACCGTGC CCAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCT CTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCG GACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGC CACGAGGATCCTGAAGTCAAGTTCAACTGGTACGTGG ATGGCGTCGAGGTGCATAATGCCAAGACAAAACCCCG GGAGGAACAGTACAACTCAACTTATAGAGTCGTGAGC GTCCTGACCGTGCTGCATCAGGACTGGCTGAACGGCA AAGAATACAAGTGCAAAGTGTCTAATAAGGCCCTGCCT GCTCCAATCGAGAAAACAATTAGCAAGGCAAAAGGGCA GCCCAGGGAACCTCAGGTGTACACTCTGCCTCCAAGCC GCGAGGAAATGACCAAGAACCAGGTCTCCCTGACATG TCTGGTGAAAGGATTCTATCCTAGTGACATTGCCGTG GAGTGGGAATCAAATGGCCAGCCAGAGAACAATTACA AGACCACACCCCCTGTGCTGGACTCTGATGGGAGTTT CTTTCTGTATTCCAAGCTGACCGTGGATAAATCTAGA TGGCAGCAGGGAAATGTCTTTAGCTGTTCCGTGATGC ATGAGGCACTGCACAACCATTACACCCAGAAATCACTG TCACTGTCCCCAGGAAAAGGCGGGGGAGGCTCTCAGC TGCAGCTGGTCCAGAGCGGAACCGAAGTGAAGAAACC CGGCGCAAGCGTCAAAGTCTCATGCAAATCAAGCGGA TACGTCTTCACCTCTTACTATCTGGTGTGGGTCCGGC AGGCACCAGGACAGGGACTGGAGTGGATGGCCACAAT CTCTCCCGGAGACGTGAACACTAGTTACGAACAGCGA TTCCAGGGCAGAGTGACCGTCACCACAGACGCTTCAAC TAATACCGTGGATATGGAGCTGCGGAGCCTGAGATCC GAAGATACAGCCGTCTACTATTGCGCTAGGGGGCCCC GCAGCAAGCCTCCTTATCTGTATTTCGCTCTGGATG TCTGGGGGCAGGGAACAGCAGTCACCGTCTCAAGCGG CGGGGGAGGCTCTGGGGGAGGCGGGAGTGGAGGCGG GGGATCAGAGATTGTCCTGACCCAGTCACCTGGCACCC TGAGCCTGAGTCCTGGAGAGACCGCTATTCTGTCTTG TCGGGCATCACAGTCCGTGAGCTCCTCTCTGCTGGCA TGGTACCAGCAGAAGCCCGGACAGGCCCCTAGGCTGC TGATCTATGGCGCCTCCAACCGCGCTACTGGCATTCG GGGGAGATTCAGTGGCTCAGGGAGCGGAACCGACTTT ACCCTGACAATCAGCCGGCTGGAGCCCGAAGATTTCGT GCTGTACTATTGTCAGCATTATGGAAGCAGGGTCAC CTTCGGACAGGGAACTAAACTGGAAATCAAG 171 Bs3GC2b heavy EVQLVESGGDLVKAGGSLRLSCAVSGLSFSSSGMNWVRQ chain aa APGKGLEWISSISGSQNYKYYADSVKGRFVVSRDNARNFL YLQMDSLRAEDTAVYFCVGGFPYWLPPSDFSGFHVWGQ GTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP SSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGKGGGGSQLQLVQSGTEV KKPGASVKVSCKSSGYVFTSYYLVWVRQAPGQGLEWMA TISPGDVNTSYEQRFQGRVTVTTDASTNTVDMELRSLRS EDTAVYYCARGPRSKPPYLYFALDVWGQGTAVTVSSGG GGSGGGGSGGGGSEIVLTQSPGTLSLSPGETAILSCRASQS VSSSLLAWYQQKPGQAPRLLIYGASNRATGIRGRFSGSGS GTDFTLTISRLEPEDFVLYYCQHYGSRVTFGQGTKLEIK 172 Bs3GC2b heavy GAGGTACAATTGGTGGAGTCTGGGGGAGACCTGGTC chain nucl AAGGCGGGGGGGTCCCTGAGACTCTCCTGTGCCGTCT CTGGATTGTCCTTCAGTAGTTCAGGCATGAATTGGGT CCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCTCA TCGATTAGTGGTAGTCAGAACTACAAATACTATGCAG ACTCAGTGAAGGGCCGATTCGTCGTCTCCAGAGACAAC GCCCGCAACTTTCTATATCTGCAAATGGACAGCCTGAG GGCCGAGGATACGGCTGTGTATTTTTGTGTGGGAGG TTTCCCCTATTGGTTACCCCCGAGCGACTTCTCCGGTT TCCATGTCTGGGGCCAAGGGACCACGGTCACCGTCTCC TCAGCGTCGACCAAGGGCCCATCGGTCTTCCCCCTGGC ACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCC TGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGT GACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGC GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACT CTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCA GCTTGGGCACCCAGACCTACATCTGCAACGTGAATCAC AAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCC CAAATCTTGTGACAAAACTCACACATGCCCACCGTGCC CAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTC TTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGG ACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCC ACGAGGATCCTGAAGTCAAGTTCAACTGGTACGTGGA TGGCGTCGAGGTGCATAATGCCAAGACAAAACCCCGG GAGGAACAGTACAACTCAACTTATAGAGTCGTGAGCG TCCTGACCGTGCTGCATCAGGACTGGCTGAACGGCAA AGAATACAAGTGCAAAGTGTCTAATAAGGCCCTGCCT GCTCCAATCGAGAAAACAATTAGCAAGGCAAAAGGGCA GCCCAGGGAACCTCAGGTGTACACTCTGCCTCCAAGCC GCGAGGAAATGACCAAGAACCAGGTCTCCCTGACATG TCTGGTGAAAGGATTCTATCCTAGTGACATTGCCGTG GAGTGGGAATCAAATGGCCAGCCAGAGAACAATTACA AGACCACACCCCCTGTGCTGGACTCTGATGGGAGTTT CTTTCTGTATTCCAAGCTGACCGTGGATAAATCTAGA TGGCAGCAGGGAAATGTCTTTAGCTGTTCCGTGATGC ATGAGGCACTGCACAACCATTACACCCAGAAATCACTG TCACTGTCCCCAGGAAAAGGCGGGGGAGGCTCTCAGC TGCAGCTGGTCCAGAGCGGAACCGAAGTGAAGAAACC CGGCGCAAGCGTCAAAGTCTCATGCAAATCAAGCGGA TACGTCTTCACCTCTTACTATCTGGTGTGGGTCCGGC AGGCACCAGGACAGGGACTGGAGTGGATGGCCACAAT CTCTCCCGGAGACGTGAACACTAGTTACGAACAGCGA TTCCAGGGCAGAGTGACCGTCACCACAGACGCTTCAAC TAATACCGTGGATATGGAGCTGCGGAGCCTGAGATCC GAAGATACAGCCGTCTACTATTGCGCTAGGGGGCCCC GCAGCAAGCCTCCTTATCTGTATTTCGCTCTGGATG TCTGGGGGCAGGGAACAGCAGTCACCGTCTCAAGCGG CGGGGGAGGCTCTGGGGGAGGCGGGAGTGGAGGCGG GGGATCAGAGATTGTCCTGACCCAGTCACCTGGCACCC TGAGCCTGAGTCCTGGAGAGACCGCTATTCTGTCTTG TCGGGCATCACAGTCCGTGAGCTCCTCTCTGCTGGCA TGGTACCAGCAGAAGCCCGGACAGGCCCCTAGGCTGC TGATCTATGGCGCCTCCAACCGCGCTACTGGCATTCG GGGGAGATTCAGTGGCTCAGGGAGCGGAACCGACTTT ACCCTGACAATCAGCCGGCTGGAGCCCGAAGATTTCGT GCTGTACTATTGTCAGCATTATGGAAGCAGGGTCAC CTTCGGACAGGGAACTAAACTGGAAATCAAG 173 Bs3GC3b heavy EVQLVESGGDLVKAGGSLRLSCAVSGLSFSSSGMNWVRQ chain aa APGKGLEWISSISGSQNYKYYADSVKGRFVVSRDNARNFL YLQMDSLRAEDTAVYFCVGGFPYWLPPSDFSGFHVWGQ GTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDY FPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP SSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPPC PAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHE DPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLT VLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREP QVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNG QPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFS CSVMHEALHNHYTQKSLSLSPGKGGGGSGVQLVQSGGGL VQPGGSLRLSCAASGFTVSTNYMSWVRQAPGKGLEWVSI LYAGGVTRYADSVKTRFTISRDNSKNTLFLQMNALSAED TAIYYCAKHYDSGYSTIDHFDSWGQGTLVTVSSGGGGSG GGGSGGGGSDIQMTQSPDSVAVSLGERATINCKSSQSVFY TSKNKNYLAWFQQKPGQPPKLLIYWASTRESGVPDRFSG SGSGTDFTLTISSLRPEDVAVYYCQQYYSTPFTFGPGTKV DIK 174 Bs3GC3b heavy GAGGTACAATTGGTGGAGTCTGGGGGAGACCTGGTC chain nucl AAGGCGGGGGGGTCCCTGAGACTCTCCTGTGCCGTCT CTGGATTGTCCTTCAGTAGTTCAGGCATGAATTGGGT CCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGATCTCA TCGATTAGTGGTAGTCAGAACTACAAATACTATGCAG ACTCAGTGAAGGGCCGATTCGTCGTCTCCAGAGACAAC GCCCGCAACTTTCTATATCTGCAAATGGACAGCCTGAG GGCCGAGGATACGGCTGTGTATTTTTGTGTGGGAGG TTTCCCCTATTGGTTACCCCCGAGCGACTTCTCCGGTT TCCATGTCTGGGGCCAAGGGACCACGGTCACCGTCTCC TCAGCGTCGACCAAGGGCCCATCGGTCTTCCCCCTGGC ACCCTCCTCCAAGAGCACCTCTGGGGGCACAGCGGCCC TGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCTGT GACGGTCTCGTGGAACTCAGGCGCCCTGACCAGCGGC GTGCACACCTTCCCGGCTGTCCTACAGTCCTCAGGACT CTACTCCCTCAGCAGCGTGGTGACCGTGCCCTCCAGCA GCTTGGGCACCCAGACCTACATCTGCAACGTGAATCAC AAGCCCAGCAACACCAAGGTGGACAAGAGAGTTGAGCC CAAATCTTGTGACAAAACTCACACATGCCCACCGTGCC CAGCACCTGAACTCCTGGGGGGACCGTCAGTCTTCCTC TTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGG ACCCCTGAGGTCACATGCGTGGTGGTGGACGTGAGCC ACGAGGATCCTGAAGTCAAGTTCAACTGGTACGTGGA TGGCGTCGAGGTGCATAATGCCAAGACAAAACCCCGG GAGGAACAGTACAACTCAACTTATAGAGTCGTGAGCG TCCTGACCGTGCTGCATCAGGACTGGCTGAACGGCAA AGAATACAAGTGCAAAGTGTCTAATAAGGCCCTGCCT GCTCCAATCGAGAAAACAATTAGCAAGGCAAAAGGGCA GCCCAGGGAACCTCAGGTGTACACTCTGCCTCCAAGCC GCGAGGAAATGACCAAGAACCAGGTCTCCCTGACATG TCTGGTGAAAGGATTCTATCCTAGTGACATTGCCGTG GAGTGGGAATCAAATGGCCAGCCAGAGAACAATTACA AGACCACACCCCCTGTGCTGGACTCTGATGGGAGTTT CTTTCTGTATTCCAAGCTGACCGTGGATAAATCTAGA TGGCAGCAGGGAAATGTCTTTAGCTGTTCCGTGATGC ATGAGGCACTGCACAACCATTACACCCAGAAATCACTG TCACTGTCCCCAGGAAAAGGCGGGGGAGGCTCTGGCG TGCAGCTGGTCCAGAGCGGAGGCGGACTGGTCCAGCC CGGCGGATCACTGAGACTGTCATGTGCCGCAAGCGGG TTTACCGTCTCTACAAACTACATGTCTTGGGTGAGGC AGGCACCTGGAAAGGGACTGGAGTGGGTCTCAATCCT GTACGCTGGCGGGGTGACCCGGTATGCAGACAGCGT CAAGACCCGGTTCACAATTAGCAGAGATAACTCCAAAA ATACTCTGTTTCTGCAGATGAATGCCCTGTCCGCTGA AGACACCGCAATCTACTATTGCGCCAAACACTATGATA GTGGGTACAGTACCATTGACCATTTCGATAGCTGGG GGCAGGGGACTCTGGTGACCGTCTCATCAGGCGGGGG AGGCTCTGGGGGAGGCGGGAGTGGAGGCGGGGGATC AGATATTCAGATGACCCAGAGTCCTGATTCCGTCGCT GTCTCACTGGGAGAAAGGGCAACCATTAACTGTAAAA GCTCACAGAGTGTCTTCTACACCAGTAAGAACAAAAA CTATCTGGCCTGGTTTCAGCAGAAGCCAGGCCAGCCCC CTAAACTGCTGATCTACTGGGCTAGCACTAGAGAGTC TGGAGTGCCAGACAGATTCTCTGGCAGTGGGTCAGGA ACCGACTTCACCCTGACAATTAGCTCCCTGAGGCCCGA AGACGTGGCCGTCTACTATTGTCAGCAGTATTATTCA ACACCCTTCACATTCGGACCAGGAACAAAAGTGGATA TTAAG 175 Bs3GC4 heavy QVQLMESGGGVVQPGRSLRLSCSAFGFTFSNYPMHWVR chain aa QAPGKGLEWVAIILPDGNRKNYGRSVTGRFTISRDNSNN SLYLQMNNLTTEDTAMYYCTRDGTYYSNGGVYQTYRR FFDFWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSQLQLV QSGTEVKKPGASVKVSCKSSGYVFTSYYLVWVRQAPGQG LEWMATISPGDVNTSYEQRFQGRVTVTTDASTNTVDME LRSLRSEDTAVYYCARGPRSKPPYLYFALDVWGQGTAVT VSSGGGGSGGGGSGGGGSEIVLTQSPGTLSLSPGETAILS CRASQSVSSSLLAWYQQKPGQAPRLLIYGASNRATGIRGR FSGSGSGTDFTLTISRLEPEDFVLYYCQHYGSRVTFGQGT KLEIK 176 BsGC4 heavy CAGGTGCAATTGATGGAGTCTGGGGGAGGCGTGGTC chain nucl CAGCCTGGGAGGTCCCTGCGACTCTCATGCAGTGCCT TTGGATTCACCTTTTCGAACTATCCTATGCACTGGGT CCGCCAGGCTCCAGGCAAGGGACTTGAGTGGGTGGCT ATCATTTTACCTGATGGGAACAGAAAAAACTATGGAA GGTCCGTGACGGGCCGATTCACCATCTCCAGAGACAAT TCCAACAACAGCCTTTATTTGCAAATGAACAACCTGAC GACTGAGGACACGGCTATGTACTATTGTACGAGAGAT GGCACGTATTACTCTAATGGTGGTGTTTATCAGACAT ATCGAAGGTTCTTCGATTTCTGGGGCCGTGGCACCCT GGTCACCGTCTCCTCAGCGTCGACCAAGGGCCCATCGG TCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCTGGG GGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACT TCCCCGAACCTGTGACGGTCTCGTGGAACTCAGGCGCC CTGACCAGCGGCGTGCACACCTTCCCGGCTGTCCTACA GTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCG TGCCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGC AACGTGAATCACAAGCCCAGCAACACCAAGGTGGACAA GAGAGTTGAGCCCAAATCTTGTGACAAAACTCACACAT GCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCG TCAGTCTTCCTCTTCCCCCCAAAACCCAAGGACACCCTC ATGATCTCCCGGACCCCTGAGGTCACATGCGTGGTGG TGGACGTGAGCCACGAGGATCCTGAAGTCAAGTTCAA CTGGTACGTGGATGGCGTCGAGGTGCATAATGCCAAG ACAAAACCCCGGGAGGAACAGTACAACTCAACTTATAG AGTCGTGAGCGTCCTGACCGTGCTGCATCAGGACTGG CTGAACGGCAAAGAATACAAGTGCAAAGTGTCTAATA AGGCCCTGCCTGCTCCAATCGAGAAAACAATTAGCAAG GCAAAAGGGCAGCCCAGGGAACCTCAGGTGTACACTC TGCCTCCAAGCCGCGAGGAAATGACCAAGAACCAGGTC TCCCTGACATGTCTGGTGAAAGGATTCTATCCTAGTG ACATTGCCGTGGAGTGGGAATCAAATGGCCAGCCAGA GAACAATTACAAGACCACACCCCCTGTGCTGGACTCTG ATGGGAGTTTCTTTCTGTATTCCAAGCTGACCGTGGA TAAATCTAGATGGCAGCAGGGAAATGTCTTTAGCTGT TCCGTGATGCATGAGGCACTGCACAACCATTACACCCA GAAATCACTGTCACTGTCCCCAGGAAAAGGCGGGGG AGGCTCTCAGCTGCAGCTGGTCCAGAGCGGAACC GAAGTGAAGAAACCCGGCGCAAGCGTCAAAGTCT CATGCAAATCAAGCGGATACGTCTTCACCTCTTA CTATCTGGTGTGGGTCCGGCAGGCACCAGGACAG GGACTGGAGTGGATGGCCACAATCTCTCCCGGA GACGTGAACACTAGTTACGAACAGCGATTCCAGG GCAGAGTGACCGTCACCACAGACGCTTCAACTAAT ACCGTGGATATGGAGCTGCGGAGCCTGAGATCCG AAGATACAGCCGTCTACTATTGCGCTAGGGGGCC CCGCAGCAAGCCTCCTTATCTGTATTTCGCTCT GGATGTCTGGGGGCAGGGAACAGCAGTCACCGT CTCAAGCGGCGGGGGAGGCTCTGGGGGAGGCGG GAGTGGAGGCGGGGGATCAGAGATTGTCCTGAC CCAGTCACCTGGCACCCTGAGCCTGAGTCCTGGA GAGACCGCTATTCTGTCTTGTCGGGCATCACAGT CCGTGAGCTCCTCTCTGCTGGCATGGTACCAGCA GAAGCCCGGACAGGCCCCTAGGCTGCTGATCTAT GGCGCCTCCAACCGCGCTACTGGCATTCGGGGGA GATTCAGTGGCTCAGGGAGCGGAACCGACTTTAC CCTGACAATCAGCCGGCTGGAGCCCGAAGATTTC GTGCTGTACTATTGTCAGCATTATGGAAGCAGG GTCACCTTCGGACAGGGAACTAAACTGGAAATCA AG 177 Bs3GC5 heavy QVQLMESGGGVVQPGRSLRLSCSAFGFTFSNYPMHWVR chain aa QAPGKGLEWVAIILPDGNRKNYGRSVTGRFTISRDNSNN SLYLQMNNLTTEDTAMYYCTRDGTYYSNGGVYQTYRR FFDFWGRGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAAL GCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCV VVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTY RVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAK GQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQ QGNVFSCSVMHEALHNHYTQKSLSLSPGKGGGGSGVQLV QSGGGLVQPGGSLRLSCAASGFTVSTNYMSWVRQAPGK GLEWVSILYAGGVTRYADSVKTRFTISRDNSKNTLFLQM NALSAEDTAIYYCAKHYDSGYSTIDHFDSWGQGTLVTVS SGGGGSGGGGSGGGGSDIQMTQSPDSVAVSLGERATINC KSSQSVFYTSKNKNYLAWFQQKPGQPPKLLIYWASTRES GVPDRFSGSGSGTDFTLTISSLRPEDVAVYYCQQYYSTPF TFGPGTKVDIK 178 Bs3GC5 heavy CAGGTGCAATTGATGGAGTCTGGGGGAGGCGTG chain nucl GTCCAGCCTGGGAGGTCCCTGCGACTCTCATGCA GTGCCTTTGGATTCACCTTTTCGAACTATCCTAT GCACTGGGTCCGCCAGGCTCCAGGCAAGGGACTT GAGTGGGTGGCTATCATTTTACCTGATGGGAACA GAAAAAACTATGGAAGGTCCGTGACGGGCCGATT CACCATCTCCAGAGACAATTCCAACAACAGCCTTT ATTTGCAAATGAACAACCTGACGACTGAGGACAC GGCTATGTACTATTGTACGAGAGATGGCACGTAT TACTCTAATGGTGGTGTTTATCAGACATATCGAA GGTTCTTCGATTTCTGGGGCCGTGGCACCCTGGT CACCGTCTCCTCAGCGTCGACCAAGGGCCCATCGG TCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTCT GGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGG ACTACTTCCCCGAACCTGTGACGGTCTCGTGGAAC TCAGGCGCCCTGACCAGCGGCGTGCACACCTTCCC GGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCA GCAGCGTGGTGACCGTGCCCTCCAGCAGCTTGGG CACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCC CAAATCTTGTGACAAAACTCACACATGCCCACCGT GCCCAGCACCTGAACTCCTGGGGGGACCGTCAGT CTTCCTCTTCCCCCCAAAACCCAAGGACACCCTCAT GATCTCCCGGACCCCTGAGGTCACATGCGTGGTG GTGGACGTGAGCCACGAGGATCCTGAAGTCAAGT TCAACTGGTACGTGGATGGCGTCGAGGTGCATAA TGCCAAGACAAAACCCCGGGAGGAACAGTACAACT CAACTTATAGAGTCGTGAGCGTCCTGACCGTGCT GCATCAGGACTGGCTGAACGGCAAAGAATACAAG TGCAAAGTGTCTAATAAGGCCCTGCCTGCTCCAA TCGAGAAAACAATTAGCAAGGCAAAAGGGCAGCC CAGGGAACCTCAGGTGTACACTCTGCCTCCAAGCC GCGAGGAAATGACCAAGAACCAGGTCTCCCTGAC ATGTCTGGTGAAAGGATTCTATCCTAGTGACATT GCCGTGGAGTGGGAATCAAATGGCCAGCCAGAGA ACAATTACAAGACCACACCCCCTGTGCTGGACTCT GATGGGAGTTTCTTTCTGTATTCCAAGCTGACCG TGGATAAATCTAGATGGCAGCAGGGAAATGTCTT TAGCTGTTCCGTGATGCATGAGGCACTGCACAAC CATTACACCCAGAAATCACTGTCACTGTCCCCAGG AAAAGGCGGGGGAGGCTCTGGCGTGCAGCTGGTCCA GAGCGGAGGCGGACTGGTCCAGCCCGGCGGATCACTG AGACTGTCATGTGCCGCAAGCGGGTTTACCGTCTCTA CAAACTACATGTCTTGGGTGAGGCAGGCACCTGGAAA GGGACTGGAGTGGGTCTCAATCCTGTACGCTGGCGG GGTGACCCGGTATGCAGACAGCGTCAAGACCCGGTTC ACAATTAGCAGAGATAACTCCAAAAATACTCTGTTTCT GCAGATGAATGCCCTGTCCGCTGAAGACACCGCAATCT ACTATTGCGCCAAACACTATGATAGTGGGTACAGTA CCATTGACCATTTCGATAGCTGGGGGCAGGGGACTC TGGTGACCGTCTCATCAGGCGGGGGAGGCTCTGGGG GAGGCGGGAGTGGAGGCGGGGGATCAGATATTCAGA TGACCCAGAGTCCTGATTCCGTCGCTGTCTCACTGGG AGAAAGGGCAACCATTAACTGTAAAAGCTCACAGAGT GTCTTCTACACCAGTAAGAACAAAAACTATCTGGCCT GGTTTCAGCAGAAGCCAGGCCAGCCCCCTAAACTGCTG ATCTACTGGGCTAGCACTAGAGAGTCTGGAGTGCCAG ACAGATTCTCTGGCAGTGGGTCAGGAACCGACTTCAC CCTGACAATTAGCTCCCTGAGGCCCGAAGACGTGGCCG TCTACTATTGTCAGCAGTATTATTCAACACCCTTCAC ATTCGGACCAGGAACAAAAGTGGATATTAAG Non-engineered chains of multispecific antibodies 179 GCA7 heavy chain GVQLVQSGGGLVQPGGSLRLSCAASgftvstnyMSWVRQAP aa GKGLEWVSIlyaggvtRYADSVKTRFTISRDNSKNTLFLQM NALSAEDTAIYYCakhydsgystidhfdsWGQGTLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSG ALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNV NHKPSNTKVDKRVEPKSCDKTHTCPPCPAPELLGGPSVFL FPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDG VEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMT KNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVL DSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHY TQKSLSLSPGK 180 GCA7 heavy chain ggggtgcaactggtgcagtctgggggaggcttggtccagccgggggggtccctgaga nucl ctctcctgtgcagcctctGGATTCACCGTCAGTACCAACTACatga gctgggtccgccaggctccagggaaggggctggagtgggtctcaattCTTTAT GCCGGAGGTGTCACAaggtacgcagactccgtgaagaccagattcaccat ctccagagacaattccaagaacactctctttcttcaaatgaacgccctgagcgccgaggac acggctatatattactgtGCGAAACACTATGATTCGGGATATTC TACCATAGATCACTTTGACTCCtggggccagggaaccctggtcacc gtctcctcagcgtcgaccaagggcccatcggtcttccccctggcaccctcctccaagagca cctctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaacctgtg acggtctcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtccta cagtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggc acccagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagaga gttgagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcct ggggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccgg acccctgaggtcacatgcgtggtggtggacgtgagccacgaGgaTcctgaggtcaag ttcaactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggagg agcagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggc tgaatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgaga aaaccatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccat cccgggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatc ccagcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagac cacgcctcccgtgctggactccgacggctccttcttcctctatagcaagctcaccgtggaca agagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcac aaccactacacgcagaagagcctctccctgtccccgggtaaa 181 GCA7 light chain DIQMTQSPDSVAVSLGERATINCKSSqsvfytsknknyLAWFQ aa QKPGQPPKLLIYwasTRESGVPDRFSGSGSGTDFTLTISSL RPEDVAVYYCqqyystpftFGPGTKVDIKRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQ ESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQG LSSPVTKSFNRGEC 182 GCA7 light chain gacatccagatgacccagtctccagactccgtggctgtgtctctgggcgagagggccacc nucl atcaactgcaagtccagcCAGAGTGTTTTCTACACCTCCAAAAA TAAAAACTACttagcttggttccagcagaaaccaggacagcctcctaaactgctc atttacTGGGCATCTacccgggagtccggggtccctgaccgattcagtggcag cgggtctgggacagatttcactctcaccatcagcagcctgcggcctgaagatgtggcagt ttattactgtCAGCAATATTATAGTACCCCTTTCACTttcggccc tgggaccaaagtggatatcaaacgTacGgtggctgcaccatctgtcttcatcttcccgcc atctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatc ccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccag gagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgac gctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagg gcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt 183 GCA21 light chain DIQMTQSPSTLSTSVGDRVTITCRASqnilnwLAWYQQKPG aa NAPNLLIYkasDLQSGVPSRFSGSGSGTEFTLTISSLQPDDF ATYYCqhynsypltFGGGTKVEIKRTVAAPSVFIFPPSDEQLK SGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTE QDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPV TKSFNRGEC 184 GCA21 light chain gacatccagatgacccagtctccttccaccctgtctacatctgtgggagacagagtcaccat nucl cacttgccgggccagtCAGAATATCCTTAATTGGttggcctggtatcaa cagaaaccagggaacgcccctaacctcctgatatatAAGGCGTCTgatttacaaa gtggggtcccctcaagattcagcggcagtgggtctgggacagaattcactctcaccatca gcagcctgcagcctgatgattttgcaacttattactgcCAGCATTATAATAG TTATCCTCTCACTttcggcggagggaccaaggtggaaatcaaacgTacG gtggctgcaccatctgtcttcatcttcccgccatctgatgagcagttgaaatctggaactgc ctctgttgtgtgcctgctgaataacttctatcccagagaggccaaagtacagtggaaggtg gataacgccctccaatcgggtaactcccaggagagtgtcacagagcaggacagcaagga cagcacctacagcctcagcagcaccctgacgctgagcaaagcagactacgagaaacacaa agtctacgcctgcgaagtcacccatcagggcctgagctcgcccgtcacaaagagcttcaa caggggagagtgt 185 GCB59 light chain SYVLTQPPSVSVAPGQTASLTCGGTNIGSKSVHWYQQKA aa GQAPVLVVYADNDRPSGVPERFSGSNSGNTATLTISRVEA EDESDYFCQVWDGNTDHVVFGGGTKLTVLGQPKAAPS VTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSP VKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQ VTHEGSTVEKTVAPTECS 186 GCB59 light chain TCATATGTGCTGACTCAACCACCCTCGGTGTCAGTGGC nucl CCCAGGACAGACGGCCAGTCTAACCTGTGGGGGAACT AACATTGGAAGTAAAAGTGTTCATTGGTACCAGCAAA AGGCAGGCCAGGCCCCTGTGTTGGTCGTCTATGCTGA TAACGACAGGCCCTCAGGGGTCCCTGAGCGATTCTCT GGCTCCAACTCTGGGAACACGGCCACCCTGACCATCAG CAGGGTCGAGGCCGAGGATGAGTCCGACTATTTCTGT CAGGTGTGGGATGGTAATACTGATCATGTGGTCTT CGGCGGAGGGACCAAGCTGACCGTCCTGggtcagcccaaggct gccccctcggtcactctgttcccgccctcctctgaggagcttcaagccaacaaggccacact ggtgtgtctcataagtgacttctacccgggagccgtgacagtggcttggaaagcagata gcagccccgtcaaggcgggagtggagaccaccacaccctccaaacaaagcaacaacaag tacgcggccagcagctatctgagcctgacgcctgagcagtggaagtcccacagaagcta cagctgccaggtcacgcatgaagggagcaccgtggagaagacagtggcccctacagaa tgttca 187 GCE536 heavy QLQLVQSGTEVKKPGASVKVSCKSSGYVFTSYYLVWVRQ chain aa APGQGLEWMATISPGDVNTSYEQRFQGRVTVTTDASTN TVDMELRSLRSEDTAVYYCARGPRSKPPYLYFALDVWG QGTAVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKD YFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTV PSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHTCPP CPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVL TVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPRE PQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESN GQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVF SCSVMHEALHNHYTQKSLSLSPGK 188 GCE536 heavy CAGCTGCAGCTGGTCCAGTCAGGCACAGAGGTCAAAA chain nucl AGCCAGGAGCATCAGTGAAGGTGTCTTGTAAGTCATC AGGATACGTGTTCACCTCTTACTATCTGGTGTGGGT CCGGCAGGCACCAGGACAGGGACTGGAGTGGATGGCC ACAATCTCTCCCGGAGACGTGAACACTAGTTACGAAC AGCGATTCCAGGGCAGAGTGACCGTCACCACAGACGCT TCAACTAATACCGTGGATATGGAGCTGCGGAGCCTGA GATCCGAAGATACAGCCGTCTACTATTGCGCTAGGGG GCCCCGCAGCAAGCCTCCTTATCTGTATTTTGCTCT GGATGTGTGGGGGCAGGGGACCGCTGTCACCGTGTC AAGCgcgtcgaccaagggcccatcggtcttccccctggcaccctcctccaagagcacc tctgggggcacagcggccctgggctgcctggtcaaggactacttccccgaacctgtgac ggtctcgtggaactcaggcgccctgaccagcggcgtgcacaccttcccggctgtcctaca gtcctcaggactctactccctcagcagcgtggtgaccgtgccctccagcagcttgggcacc cagacctacatctgcaacgtgaatcacaagcccagcaacaccaaggtggacaagagagtt gagcccaaatcttgtgacaaaactcacacatgcccaccgtgcccagcacctgaactcctgg ggggaccgtcagtcttcctcttccccccaaaacccaaggacaccctcatgatctcccggacc cctgaggtcacatgcgtggtggtggacgtgagccacgaGgaTcctgaggtcaagttc aactggtacgtggacggcgtggaggtgcataatgccaagacaaagccgcgggaggag cagtacaacagcacgtaccgtgtggtcagcgtcctcaccgtcctgcaccaggactggctg aatggcaaggagtacaagtgcaaggtctccaacaaagccctcccagcccccatcgagaaa accatctccaaagccaaagggcagccccgagaaccacaggtgtacaccctgcccccatccc gggaggagatgaccaagaaccaggtcagcctgacctgcctggtcaaaggcttctatccc agcgacatcgccgtggagtgggagagcaatgggcagccggagaacaactacaagacca cgcctcccgtgctggactccgacggctccttcttcctctatagcaagctcaccgtggacaa gagcaggtggcagcaggggaacgtcttctcatgctccgtgatgcatgaggctctgcaca accactacacgcagaagagcctctccctgtccccgggtaaa 189 GCE536 light EIVLTQSPGTLSLSPGETAILSCRASQSVSSSLLAWYQQKP chain aa GQAPRLLIYGASNRATGIRGRFSGSGSGTDFTLTISRLEPE DFVLYYCQHYGSRVTFGQGTKLEIKRTVAAPSVFIFPPSD EQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQE SVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGL SSPVTKSFNRGEC 190 GCE536 light gaaattgtgttgacgcagtctcctggcaccctgtctttgtctccaggggaaacagccatcct chain nucl ctcctgcagggccagtcagagtgtcagcagcagcctcttagcctggtaccagcaaaaa cctggccaggctcccaggctcctcatctacggtgcatccaatagggccactggcatcaga ggcaggtttagtggcagtgggtctgggacagacttcactctcaccatcagtagattggag cctgaagattttgtactttattactgtcagcactatggctcacgggtcacttttggccag gggaccaagctggagatcaaacgTacGgtggctgcaccatctgtcttcatcttcccgcc atctgatgagcagttgaaatctggaactgcctctgttgtgtgcctgctgaataacttctatc ccagagaggccaaagtacagtggaaggtggataacgccctccaatcgggtaactcccag gagagtgtcacagagcaggacagcaaggacagcacctacagcctcagcagcaccctgac gctgagcaaagcagactacgagaaacacaaagtctacgcctgcgaagtcacccatcagg gcctgagctcgcccgtcacaaagagcttcaacaggggagagtgt *the sequences highlighted in bold are CDR regions (nucleotide or aa) and the underlined residues are mutated residues as compared to the ″germline″ sequence.