HUMAN TUMOR NECROSIS FACTOR ALPHA ANTIBODIES

Abstract

The present disclosure relates to antibodies that specifically bind soluble and membrane forms of human TNFα, compositions comprising such TNFα antibodies, and methods of using such TNFα antibodies and compositions.

Claims

1. An antibody that binds human TNFα, wherein the antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein: the HCDR1 comprises SEQ ID NO: 1; the HCDR2 comprises SEQ ID NO: 2; the HCDR3 comprises SEQ ID NO: 3; the LCDR1 comprises SEQ ID NO: 4; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 6.

2. The antibody claim 1, wherein the VH comprises SEQ ID NO: 7 and the VL comprises SEQ ID NO: 8.

3. The antibody of claim 1, wherein the antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises SEQ ID NO: 9 and the LC comprises SEQ ID NO: 10.

4. An antibody that binds human TNFα, wherein the antibody comprises a heavy chain variable region (VH) and a light chain variable region (VL), wherein the VH comprises heavy chain complementarity determining regions HCDR1, HCDR2, and HCDR3, and the VL comprises light chain complementarity determining regions LCDR1, LCDR2, and LCDR3, wherein: a. the HCDR1 comprises SEQ ID NO: 22; the HCDR2 comprises SEQ ID NO: 23; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 4, 14, or 46; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 6; b. the HCDR1 comprises SEQ ID NO: 22; the HCDR2 comprises SEQ ID NO: 23; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 14; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 15 or 47; c. the HCDR1 comprises SEQ ID NO: 1; the HCDR2 comprises SEQ ID NO: 2; the HCDR3 comprises SEQ ID NO: 30; the LCDR1 comprises SEQ ID NO: 31; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 32; or d. the HCDR1 comprises SEQ ID NO: 1; the HCDR2 comprises SEQ ID NO: 2; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 14; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 15.

5. The antibody of claim 4, wherein: the HCDR1 comprises SEQ ID NO: 22; the HCDR2 comprises SEQ ID NO: 23; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 4; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 6.

6. The antibody of claim 4, wherein: the HCDR1 comprises SEQ ID NO: 22; the HCDR2 comprises SEQ ID NO: 23; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 14; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 6.

7. The antibody of claim 4, wherein: the HCDR1 comprises SEQ ID NO: 22; the HCDR2 comprises SEQ ID NO: 23; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 14; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 15.

8. The antibody of claim 4, wherein: the HCDR1 comprises SEQ ID NO: 22; the HCDR2 comprises SEQ ID NO: 23; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 46; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 6.

9. The antibody of claim 4, wherein: the HCDR1 comprises SEQ ID NO: 22; the HCDR2 comprises SEQ ID NO: 23; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 14; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 47.

10. The antibody of claim 4, wherein the VH comprises SEQ ID NO: 24 and the VL comprises SEQ ID NO: 8, 17, or 27.

11. The antibody of claim 4, wherein the antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises SEQ ID NO: 25 and the LC comprises SEQ ID NO: 10, 19, or 28.

12. The antibody of claim 4, wherein: the HCDR1 comprises SEQ ID NO: 1; the HCDR2 comprises SEQ ID NO: 2; the HCDR3 comprises SEQ ID NO: 30; the LCDR1 comprises SEQ ID NO: 31; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 32.

13. The antibody of claim 12, wherein the VH comprises SEQ ID NO: 33 and the VL comprises SEQ ID NO: 34.

14. The antibody of claim 12, wherein the antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises SEQ ID NO: 35 and the LC comprises SEQ ID NO: 36.

15. The antibody of claim 4, wherein: the HCDR1 comprises SEQ ID NO: 1; the HCDR2 comprises SEQ ID NO: 2; the HCDR3 comprises SEQ ID NO: 13; the LCDR1 comprises SEQ ID NO: 14; the LCDR2 comprises SEQ ID NO: 5; and the LCDR3 comprises SEQ ID NO: 15.

16. The antibody of claim 15, wherein the VH comprises SEQ ID NO: 16 and the VL comprises SEQ ID NO: 17.

17. The antibody of claim 15, wherein the antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC comprises SEQ ID NO: 18 and the LC comprises SEQ ID NO: 19.

18. The antibody of claim 1, wherein the antibody comprises a light chain and a heavy chain, wherein the heavy chain comprises: a cysteine at amino acid residue 124 (EU numbering); a cysteine at amino acid residue 378 (EU numbering); or a cysteine at amino acid residue 124 (EU numbering) and a cysteine at amino acid residue 378 (EU numbering).

19. The antibody of claim 4, wherein the antibody comprises a light chain and a heavy chain, wherein the heavy chain comprises: a cysteine at amino acid residue 124 (EU numbering); a cysteine at amino acid residue 378 (EU numbering); or a cysteine at amino acid residue 124 (EU numbering) and a cysteine at amino acid residue 378 (EU numbering).

20. The antibody of claim 1, wherein the antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC is human IgG1 isotype.

21. The antibody of claim 4, wherein the antibody comprises a heavy chain (HC) and a light chain (LC), wherein the HC is human IgG1 isotype.

22. A nucleic acid comprising a sequence encoding SEQ ID NO: 9, 10, 18, 19, 25, 28, 35 or 36.

23. A vector comprising the nucleic acid of claim 22.

24. The vector of claim 23, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 9, 18, 25, or 35 and a second nucleic acid sequence encoding SEQ ID NO: 10, 19, 28, or 36.

25. The vector of claim 23, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 9 and a second nucleic acid sequence encoding SEQ ID NO: 10.

26. The vector of claim 23, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 18 and a second nucleic acid sequence encoding SEQ ID NO: 19.

27. The vector of claim 23, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 25 and a second nucleic acid sequence encoding SEQ ID NO: 10, 19, or 28.

28. The vector of claim 23, wherein the vector comprises a first nucleic acid sequence encoding SEQ ID NO: 35 and a second nucleic acid sequence encoding SEQ ID NO: 36.

29. A composition comprising a first vector comprising a nucleic acid sequence encoding SEQ ID NO: 9, 18, 25, or 35 and a second vector comprising a nucleic acid sequence encoding SEQ ID NO: 10, 19, 28, or 36.

30. The composition of claim 29, wherein the first vector comprises a nucleic acid sequence encoding SEQ ID NO: 9 and the second vector comprises a nucleic acid sequence encoding SEQ ID NO: 10.

31. The composition of claim 29, wherein the first vector comprises a nucleic acid sequence encoding SEQ ID NO: 18, and the second vector comprises a nucleic acid sequence encoding SEQ ID NO: 19.

32. The composition of claim 29, wherein the first vector comprises a nucleic acid sequence encoding SEQ ID NO: 25, and the second vector comprises a nucleic acid sequence encoding SEQ ID NO: 10, 19, or. 28.

33. The composition of claim 29, wherein the first vector comprises a nucleic acid sequence encoding SEQ ID NO: 35, and the second vector comprises a nucleic acid sequence encoding SEQ ID NO: 36.

34. A cell comprising the vector of claim 23.

35. The cell of claim 34, wherein the cell is a mammalian cell.

36. A process of producing an antibody comprising culturing the cell of claim 34, under conditions such that the antibody is expressed and recovering the expressed antibody from the culture medium.

37. An antibody produced by the process of claim 36.

38. An antibody drug conjugate comprising the antibody of claim 1.

39. An antibody drug conjugate comprising the antibody of claim 4.

40. A pharmaceutical composition comprising the antibody of claim 1, and a pharmaceutically acceptable excipient, diluent, or carrier.

41. A pharmaceutical composition comprising the antibody of claim 4, and a pharmaceutically acceptable excipient, diluent, or carrier.

42. A method of treating a TNFα associated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the antibody of claim 1.

43. The method of claim 42, wherein the TNFα associated disorder is a chronic autoinflammatory immune disorder.

44. The method of claim 43, wherein the chronic autoinflammatory immune disorder is selected from Rheumatoid Arthritis (RA), Juvenile Idiopathic Arthritis, Psoriatic Arthritis (PsA), Ankylosing Spondylitis (AS), Crohn's Disease (CD), Ulcerative Colitis, Plaque Psoriasis (PS), Hidradenitis Suppurativa, Uveitis, Non-Infectious Intermediate, Posterior, Pan Uveitis, or Behcet's Disease.

45. The method claim 42, wherein the subject being administered the therapeutically effective amount of the antibody received a prior treatment with other anti-TNFα therapeutic, and wherein the subject developed anti-drug antibodies against the other anti-TNFα therapeutic.

46. The method of claim 45, wherein the other anti-TNFα therapeutic is selected from Adalimumab, Infliximab, Golimumab, Certolizumab, or Etanercept.

47. The method of claim 45, wherein the antibody has low to no binding to anti-drug antibodies against Adalimumab.

48. A method of treating a TNFα associated disorder in a subject in need thereof, comprising administering to the subject a therapeutically effective amount of the antibody of claim 4.

49. The method of claim 48, wherein the TNFα associated disorder is a chronic autoinflammatory immune disorder.

50. The method of claim 49, wherein the chronic autoinflammatory immune disorder is selected from Rheumatoid Arthritis (RA), Juvenile Idiopathic Arthritis, Psoriatic Arthritis (PsA), Ankylosing Spondylitis (AS), Crohn's Disease (CD), Ulcerative Colitis, Plaque Psoriasis (PS), Hidradenitis Suppurativa, Uveitis, Non-Infectious Intermediate, Posterior, Pan Uveitis, or Behcet's Disease.

51. The method claim 48, wherein the subject being administered the therapeutically effective amount of the antibody received a prior treatment with other anti-TNFα therapeutic, and wherein the subject developed anti-drug antibodies against the other anti-TNFα therapeutic.

52. The method of claim 51, wherein the other anti-TNFα therapeutic is selected from Adalimumab, Infliximab, Golimumab, Certolizumab, or Etanercept.

53. The method of claim 51, wherein the antibody has low to no binding to anti-drug antibodies against Adalimumab.

54. The antibody of claim 1, wherein the antibody neutralizes human TNFα.

55. The antibody of claim 4, wherein the antibody neutralizes human TNFα.

56. The antibody of claim 1, wherein the antibody is an internalizing antibody.

57. The antibody of claim 4, wherein the antibody is an internalizing antibody.

58. The antibody of claim 1, wherein the antibody has low immunogenicity.

59. The antibody of claim 4, wherein the antibody has low immunogenicity.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0089] FIG. 1 shows that exemplified anti-human TNFα antibody Ab6 has low to no binding to anti-drug antibodies against Adalimumab formed in cynomolgus monkeys hyperimmunized with Adalimumab.

[0090] FIG. 2 shows that exemplified anti-human TNFα antibody Ab6 has low to no binding to anti-drug antibodies against Adalimumab formed in human patients treated with Adalimumab.

EXAMPLES

[0091] The following examples are offered to illustrate, but not to limit, the claimed invention.

Example 1: Generation and Engineering of Anti-Human TNFα Antibodies

[0092] Antibody generation: To develop antibodies specific to human TNFα, transgenic mice with human immunoglobulin variable regions were immunized with recombinant human TNFα. Screening was done with human TNFα and the cross reactivity with other TNFα species was tested. Antibodies cross reactive to cynomolgus monkey were cloned, expressed, and purified by standard procedures, and tested for neutralization in a TNFα induced cytotoxicity assay. Antibodies were selected and engineered in their CDRs, variable domain framework regions, and IgG isotype to improve binding affinities and developability properties such as, stability, solubility, viscosity, hydrophobicity, and aggregation.

[0093] The amino acid sequence of human TNFα is provided as SEQ ID NO: 39, the amino acid sequence of cynomolgus monkey TNFα is provided as SEQ ID NO: 45.

[0094] The TNFα antibodies can be synthesized and purified by well-known methods. An appropriate host cell, such as Chinese hamster ovarian cells (CHO), can be either transiently or stably transfected with an expression system for secreting antibodies using a predetermined HC:LC vector ratio if two vectors are used, or a single vector system encoding both heavy chain and light chain. Clarified media, into which the antibody has been secreted, can be purified using the commonly used techniques.

Antibody engineering to improve viscosity: The parental TNFα antibody lineage was found to have high viscosity upon concentration. Viscosity is a key developability criteria for assessing feasibility of delivery of a therapeutic antibody via autoinjector. Mutagenesis analysis of the antibody required a fine balancing of improving biophysical properties and retaining desirable affinity and potency without increasing immunogenicity risk. In-silico modeling of the parental antibody was used to identify regions of charge imbalance in the surface comprised of the 6 complementary determining regions (CDRs). The antibodies generated from the mutagenesis were screened for TNFα binding, and those antibodies which retained or improved target binding as compared to the parental mAb (as determined by ELISA) and had desirable viscosity and other developability properties were selected for further development.
Antibody engineering to reduce the risk of immunogenicity: The exemplified anti-human TNFα antibodies were further tested in an MHC-associated peptide proteomics (MAPPS) assay to determine immunogenicity risk. Briefly, major histocompatibility complex (MHC) bound peptides were identified for antibodies with specific CDR sequences. A CDR library having mutations identified as potentially reducing immunogenicity was constructed and screened for TNFα binding. Antibodies were screened and selected to optimize by engineering for low immunogenicity risk whilst balancing maintaining desirable binding affinity to TNFα and other desirable developability properties.

[0095] Tables 1 and 2 show the exemplified anti-human TNFα antibody sequences engineered to balance reduced viscosity, low immunogenicity risk and other desirable developability properties while retaining desirable binding affinity to human TNFα.

TABLE-US-00001 TABLE 1 CDR amino acid sequences of exemplified anti-human TNFa antibodies TNFα CDR Sequence Antibody HCDR1 HCDR2 HCDR3 LCDR1 LCDR2 LCDR3 Ab1 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 1  NO: 2  NO: 3  NO: 4  NO: 5 NO: 6  Ab2 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 1  NO: 2  NO: 13 NO: 14 NO: 5 NO: 15 Ab3 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 22 NO: 23 NO: 13 NO: 4 NO: 5 NO: 6  Ab4 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 22 NO: 23 NO: 13 NO: 14 NO: 5 NO: 15 Ab5 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 22 NO: 23 NO: 13 NO: 14 NO: 5 NO: 6  Ab6 SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID SEQ ID NO: 1  NO: 2  NO: 30 NO: 31 NO: 5 NO: 32

TABLE-US-00002 TABLE 2 Amino Acid sequences of exemplified anti-human TNFα antibodies TNFα Antibody VH VL HC LC Ab1 SEQ ID SEQ ID SEQ ID SEQ ID NO: 7 NO: 8 NO: 9 NO: 10 Ab2 SEQ ID SEQ ID SEQ ID SEQ ID NO: 16 NO: 17 NO: 18 NO: 19 Ab3 SEQ ID SEQ ID SEQ ID SEQ ID NO: 24 NO: 8 NO: 25 NO: 10 Ab4 SEQ ID SEQ ID SEQ ID SEQ ID NO: 24 NO: 17 NO: 25 NO: 19 Ab5 SEQ ID SEQ ID SEQ ID SEQ ID NO: 24 NO: 27 NO: 25 NO: 28 Ab6 SEQ ID SEQ ID SEQ ID SEQ ID NO: 33 NO: 34 NO: 35 NO: 36

Example 2. Binding Affinity of Exemplified Anti-Human TNFα Antibodies

[0096] Binding Affinity, method 1: Binding affinities of the antibodies to human, rhesus macaque, mouse, rat, rabbit, and canine TNFα protein were tested in an antigen-down ELISA format. Briefly, 384-well high binding plates (Greiner Bio-one #781061) were coated with 20 μL per well of 1 μg/mL of human TNFα (Syngene), 2 μg/mL of rhesus macaque TNFα (R&D Systems, Cat #1070-RM), 2 μg/mL of mouse TNFα (R&D Systems, Cat #410-MT/CF), or 2 μg/mL of rat TNFα (R&D Systems, Cat #510-RT-CF), 2 μg/mL of rabbit TNFα (R&D Systems, Cat #5670-TG/CF), or 2 μg/mL of canine TNFα (R&D Systems, Cat #1507-CT/CF) diluted in carbonate buffer pH 9.3 (0.015 M Na.sub.2CO.sub.3 and 0.035 M NaHCO.sub.3) and stored at 4° C. overnight. Next day, the plates were blocked with 80 μL casein (Thermo Fisher Pierce, Cat #37528) for 1 h at room temperature, blocking buffer was removed, and 20 μL of titrated purified antibody expressed in CHO cells (starting concentration at 20 μg/mL diluted in casein and titrated 3-fold, 8 points down) was added to the plate. The plate was incubated at 37° C. for 90 min, then washed three times in PBS/0.1% Tween. 20 μL of secondary antibody reagent goat-anti-human-kappa-AP (Southern Biotech, Cat #2060-04) with 1:1500 dilution was added to the plate and incubated for 45 minutes at 37° C. Plates were washed 3 times in PBS/0.1% Tween, and 20 μL of alkaline phosphatase substrate solution diluted to 1:35 in molecular grade water was added to every well. Once the color developed (approximately 15-30 min), plates were read at 560 nM OD on a Molecular Device Spectramax plate reader and data was acquired using Softmax Pro 4.7 software. Data analysis was performed in GraphPad Prism.

[0097] The results as demonstrated in Table 3 show that the exemplified anti-human TNFα antibodies Ab1, Ab2, Ab3, Ab4, Ab5, and Ab6 bound human, rhesus macaque monkey, and canine TNFα with desirable affinities.

TABLE-US-00003 TABLE 3 Binding affinity of exemplified anti-human TNFα antibodies to human, rhesus macaque, and canine TNFα Human TNFα Rhesus Macaque TNFα Canine TNFα Antibody EC.sub.50 (μg/mL) EC.sub.50 (μg/mL) EC.sub.50 (μg/mL) Ab1 0.2225 0.1555 0.2204 Ab2 0.222 0.1621 0.24 Ab3 0.2164 0.1414 0.2401 Ab4 0.2134 0.1444 0.2306 Ab5 0.2201 0.1607 0.1757 Ab6 0.1313 0.1148 0.162
Binding affinity, method 2: An MSD Sector S 600 instrument (Meso Scale Discovery, Rockville, Md.) was used for reading MSD plates. Human and cynomolgus monkey TNFα were biotinylated using a Thermo-Fisher biotinylation kit. MSD assay plates were prepared as follows: a multi-array streptavidin-coated 96-well plate (Meso Scale Discovery, Cat #L15SA-1) was coated for one hour at room temperature (approximately 25° C. with 40 μL per well of 1 μg/mL solution of either biotinylated human TNFα or biotinylated cynomolgus monkey TNFα in PBS. The plates were washed 3 times in PBS +0.1% Tween (PBST) following coating, then blocked for 1 hour at room temperature with 1% bovine serum albumin (BSA) in PBS. The plates were then washed 3× with PBS prior to adding sample solutions. Solution equilibrium titration (SET) samples were prepared in 1% BSA. Anti-human TNFα antibody Ab1 was diluted to 10 μM and TNFα was serially diluted for a total of 12 dilutions. TNFα titrations and fixed antibody solutions were combined 1:1 to prepare the SET solutions. SET solutions were incubated at 37° C. for approximately 72 hours to allow binding to reach equilibrium. 40 μL of the SET solutions was transferred to the prepared MSD plate in triplicate rows and incubated at room temperature for 2.5 minutes to capture free antibody with agitation by manually tapping the plate lightly. Following incubation, the plate was washed 3 times with PBST. Then, 40 μL of 1 μg/mL SULFO-Tag anti-human/NHP kappa antibody (Meso Discovery Scale, Cat #D20TF-6) in 1% BSA was added to all wells. This was incubated statically at room temperature for one hour. The plate was then washed 3 times with PB ST, then 150 μL of MSD GOLD Read Buffer A (Meso Scale Discovery, Cat #R92TG-2) was added before reading the plate. Dilution series were prepared in triplicate in each individual experiment, and the three independent replicate experiments were conducted. The K.sub.D was determined using a quadratic kinetic model in XLfit from the MSD-SET data. Replicate K.sub.D values were entered into GraphPad Prism for both human and cynomolgus monkey TNFα to determine standard deviation statistics.

[0098] The results of this assay show that the exemplified anti-human TNFα antibody Ab1 binds to human TNFα with a K.sub.D of 8.5±1.6 μM and to cynomolgus monkey TNFα with a K.sub.D of 21.2±7.4 μM.

Example 3: Functional Activities of Exemplified Anti-Human TNFα Antibodies

[0099] Internalization of membrane bound TNFα antibodies: Internalization of exemplified anti-human TNFα antibodies upon binding to membrane expressed TNFα was tested in vitro on a CHO cell line stably transfected to express membrane human TNFα (non-cleavable TNFα). Briefly, F(ab′)2 fragment goat anti-human IgG (Jackson #109-006-098) was conjugated to pHrodo pH-sensitive dye (Fisher P36014) using manufacturer's protocol. The exemplified anti-human TNFα antibodies were incubated with equimolar amount of F(ab′)2 goat anti-hIgG-pHrodo in CHO growth media for 30 min room temp. The antibody-dye mixtures were added to CHO human TNFα transfected cells, then incubated in a 5% CO.sub.2 shaker incubator at 37° C. for 3, 6, and 24 hour time points. Final concentrations of the exemplified TNFα antibodies were 10 μg/mL and 3.3 μg/mL. Cells were washed at indicated timepoints and analyzed on a BD Fortessa flow cytometer.

[0100] The results as demonstrated in Table 4, show that the anti-human TNFα antibodies tested, were internalized into the CHO cells upon binding to membrane human TNFα expressed on CHO cells at 3.33 μg/mL and 10 μg/mL. The results further showed that the anti-human TNFα antibodies were internalized into the lysosome upon binding membrane TNFα expressed on the lysosome (data not shown).

TABLE-US-00004 TABLE 4 Internalization of exemplified anti-human TNFα antibodies Internalization Internalization Internalization Antibody at 3 hr at 6 hr at 24 hr Antibody μg/mL (Phrodo MFI) (Phrodo MFI) (Phrodo MFI) Ab1 3.33 1913 3663 1152 Ab2 3.33 1811 3266 1168 Ab3 3.33 2107 3823 1197 Ab4 3.33 2128 3518 1177 hIgG1 3.33 304 318 313 Isotype Ab1 10.0 3083 5989 4575 Ab2 10.0 2686 5158 4643 Ab3 10.0 3339 6281 4811 Ab4 10.0 2998 5750 4352 hIgG1 10.0 339 340 337 Isotype
Inhibition of soluble and membrane TNFα induced cell killing: Inhibition of soluble and membrane TNFα induced cell killing by the exemplified anti-human TNFα antibodies was evaluated in an in vitro cell based assay using L929 mouse fibrosarcoma cells which naturally express the TNF receptor. When combined with Actinomycin-D, TNFα induces classical apoptosis in these cells, resulting in rapid cell death due to excessive formation of reactive oxygen intermediates which can be rescued by TNFα neutralization. The quantity of viable cells can be measured using MTS-tetrazolium cytotoxicity assay, where the mitochondrial dehydrogenase enzymes in metabolically active cells reduces the MTS-tetrazolium into a colored formazan product, which can be detected with a microplate reader (Biotek Cytation 5 Imaging Multi-Mode Reader).

[0101] Inhibition of soluble TNFα induced cell killing: To evaluate the ability of the exemplified anti-human TNFα antibodies to inhibit soluble TNFα induced cell killing, L929 cells were treated with either human TNFα or cynomolgus monkey TNFα, separately. L929 cells resuspended at 10,000 cells/100 μL in assay medium (lx DMEM media, 10% FBS, 1% Pen-Strep, 1% MEM essential amino acids, 1% L-glutamine, 1% sodium pyruvate) were added to 96-well plates and placed in a tissue culture incubator overnight. The next day, exemplified antibodies were diluted at concentrations ranging from 15 μg/mL to 0.0005 μg/mL (with three-fold dilution), and 100 μL of each concentration of the exemplified anti-human TNFα antibodies was added in duplicate to wells containing one of the two following conditions: 200 pg/mL recombinant human TNFα, or 750 pg/mL recombinant cynomolgus TNFα, and plates were incubated for 30 min at room temperature. Human IgG1 isotype control antibody was used as a negative control. The antibody/TNFα/actinomycin-D mixture was then transferred to the 96-well plates with L929 adherent cells, and incubated in a tissue culture incubator for 18 hrs. The assay medium was removed, and 100 μL of MTS-tetrazolium substrate mixture was added to the wells and plates were incubated in a tissue culture incubator for 2 hrs. To determine cell death, plates were read at 490 nm on a microplate reader (Biotek Cytation 5 Imaging Multi-Mode Reader). Results were expressed as the concentration where 50% of the TNFα-induced cytotoxicity was inhibited (IC.sub.50, average of two independent experiments ±SEM) by the exemplified anti-human TNFα antibodies, calculated using a 3-parameter sigmoidal fit of the data (GraphPad Prism 9). The IC.sub.50 values are shown in Table 5a.

[0102] Inhibition of membrane TNFα induced cell killing: To evaluate the ability of the exemplified anti-human TNFα antibodies to inhibit membrane TNFα induced cell killing, the non-cleavable TNFα construct was stably transfected into Chinese hamster ovary (CHO) cells to generate cell surface (membrane) TNFα expressing CHO cells. The non-cleavable TNFα construct was generated with known mutations at the cleavage sites of the TNFα which allowed for expression of bioactive TNFα on the cell surface (Mueller et. al. 1999) in the absence of TNFα cleavage. Incubation of L929 cells with CHO cells expressing the human non-cleavable TNFα resulted in rapid L929 cell death. To determine whether the exemplified anti-human TNFα antibodies could neutralize the observed cell killing a dose range from 15 μg/mL to 0.0005 μg/mL (with three-fold dilution) was evaluated for each of the exemplified antibodies. Each concentration of the exemplified anti-human TNFα antibodies (100 μL/well) was added in duplicate to plates containing 500 CHO TNFα transfectant cells/well+6.25 μg/mL actinomycin-D. The antibody plus CHO cell mixtures were incubated for 30 min at room temperature and then added into the L929 cell plate. A human IgG1 isotype control antibody was used as a negative control and tested at similar dose range to the anti-human TNFα antibodies. The L929 cell death was determined essentially as described for the soluble TNFα induced cell killing assay. The IC.sub.50 values are shown in Table 5b.

[0103] The results, as demonstrated in Table 5a and 5b, showed that the exemplified anti-human TNFα antibodies inhibited soluble human TNFα or soluble cynomolgus TNFα induced L929 cell killing, and human membrane TNFα induced cell killing of L929 cells. A dose dependent inhibition of cell killing response was observed. Specifically, the IC.sub.50 for inhibition of soluble human TNFα induced cell killing (Table 5a) by the exemplified anti-human TNFα antibodies tested ranged from about 0.13 μg/mL to about 0.22 μg/mL, and from about 0.02 μg/mL to about 0.3 μg/mL for inhibition of soluble cynomolgus TNFα induced cell killing. The IC.sub.50 for inhibition of human membrane TNFα induced cell killing (Table 5b) by the exemplified anti-human TNFα antibodies tested ranged from about 0.13 μg/mL to about 0.12 μg/mL. The negative control hIgG1 isotype as expected, did not inhibit TNFα induced cell killing.

TABLE-US-00005 TABLE 5a Exemplified anti-human TNFα antibodies inhibit soluble human and soluble cynomolgus monkey TNFα induced cell killing of L929 cells Soluble Soluble cynomolgus human TNFα monkey TNFα IC.sub.50 Std. Error IC.sub.50 Std. Error Antibody (μg/mL) of Mean (μg/mL) of Mean hIgG1 Isotype n/a n/a n/a n/a Ab1 0.16 0.02 0.02 0.005 Ab2 0.13 0.03 0.02 0.004 Ab3 0.19 0.02 0.02 0.004 Ab4 0.19 0.00 0.02 0.005 Ab5 0.22 0.04 0.03 0.009

TABLE-US-00006 TABLE 5b Exemplified anti-human TNFα antibodies inhibit membrane TNFα-induced cell killing of L929 cells Antibody IC.sub.50 (μg/mL) Std. Error of Mean hIgG1 Isotype n/a n/a Ab1 0.13 0.04 Ab2 0.13 0.07 Ab3 0.15 0.08 Ab4 0.13 0.05 Ab5 0.20 0.09

Example 4: Characterization of Exemplified Anti-Human TNFα Antibodies Binding to Anti-Drug Antibodies Against Adalimumab

[0104] Binding to Cynomolgus monkey anti-drug antibodies against Adalimumab: Binding of exemplified anti-human TNFα antibodies to anti-drug antibodies against Adalimumab (anti-Adalimumab antibodies) obtained from affinity purified hyperimmune monkey serum (AP-HIMS) from cynomolgus monkeys hyperimmunized with Adalimumab was evaluated. An Adalimumab-AffiGe110 was used to purify the anti-Adalimumab antibodies from the Adalimumab hyperimmunized Cynomolgus monkeys. The anti-Adalimumab antibodies were detected using a titration of AP-HIMS in an ACE-Bridge assay. The assay was developed following the FDA Guidance on Immunogenicity testing. Briefly, streptavidin-coated 96-well plates (Pierce, 15500) were washed with 1×TBST (Boston BioProducts, IBB-181X), and coated with 30 nM biotinylated Adalimumab at 100 μL/well in TBST/0.1% bovine serum albumin (BSA; Sigma, A7888) for 1 h at room temperature. Plates were washed three times with TBST, and affinity purified anti-Adalimumab antibodies were diluted 1:10 with TBS (Fisher, BP2471-1), and added at 100 uL/well to the coated plates and incubated overnight at 4° C. The following day, plates were washed three times with TBST and the captured anti-Adalimumab antibodies were acid eluted using 65 μL/well of 300 mM acetic acid (Fisher Scientific, A38-500) for 5 min at room temperature. Polypropylene 96-well plates (Corning, 3359) were then loaded with 50 μL of 1 μg/mL each of biotinylated Adalimumab and ruthenium-labeled Adalimumab in neutralizing buffer (0.375 M Tris, 300 mM NaCl, pH 9). Next, 50 μL of the acid eluted samples were added to the polypropylene plate containing the mixture in neutralizing buffer and the ADA and were allowed to bridge to the labeled antibodies for 1 h at room temperature. MSD Gold 96-well streptavidin plates (Mesoscale, L15SA-1) were washed and blocked with TBS+1% BSA for 1 h at room temperature, then washed, and 80 μL of bridged samples were added to the plate for 1 h. The plates were washed three times with TBST, and 150 μL/well of 2×MSD Buffer (Mesoscale, R92TC-2) was added to the plates. Plates were read on an MSD SQ120 reader to provide the Tier 1 signal expressed as electro chemiluminescent units (ECLU).

[0105] The same AP-HIMS was also tested in the ACE-Bridge assay to detect antibodies against exemplified anti-human TNFα antibody Ab6, following essentially the same method outlined above for Adalimumab, but using biotin and ruthenium-labeled Ab6. The resulting ECLU signal was then plotted as a function of the concentration of AP-HEWS tested.

[0106] The results as demonstrated in FIG. 1, show that the exemplified anti-human TNFα antibody Ab6 had low to no binding to the anti-drug antibodies against Adalimumab (maximum ECLU signal of 4000) purified from serum from Cynomolgus monkey hyperimmunized with Adalimumab, when compared to binding of Adalimumab to its own anti-drug antibodies (maximum ECLU signal 40000). Specifically, the results showed that the exemplified anti-human TNFα antibody Ab6 only recognized about 10% of the anti-drug antibodies against Adalimumab raised by the cynomolgus monkeys suggesting that this binding is likely due to shared sequences located away from the CDR regions, such as the antibody constant region.

Binding to human patient anti-drug antibodies against Adalimumab: Binding of exemplified anti-human TNFα antibodies to anti-drug antibodies against Adalimumab (anti-Adalimumab antibodies) in 21 patient serum samples obtained from Adalimumab-treated patients enrolled in the study RA-BEAM was evaluated. The 21 serum samples were collected post-baseline, and were confirmed to have high ADA titers against Adalimumab, by using the methods essentially as described for the Cynomolgus monkey ADA evaluation. The 21 serum samples were then evaluated for binding to exemplified anti-human TNFα antibody Ab6 using the methods essentially as described for the Cynomolgus monkey ADA evaluation.

[0107] The results as demonstrated in FIG. 2, show that the exemplified anti-human TNFα antibody Ab6 had low to no binding to the anti-drug antibodies against Adalimumab in 16 of the 21 patient samples tested (ECLU signal was below the cut-off point of the assay (102 ECLU)). In determining immunogenicity, the cut-off point is a threshold that is used to identify “putative positive”, or anti-drug antibody containing, samples. As shown in FIG. 2, five out of 21 samples had an ECLU signal above the cut-off point, but were all less than 20% above the cut-off point, and therefore determined to be within the variability of the assay. The significantly low or no binding of anti-human TNFα antibody Ab6 to the anti-drug antibodies against Adalimumab in human patients treated with Adalimumab indicated that the Ab6 and Adalimumab antibody sequences are sufficiently different, such that, the ADA raised against Adalimumab by the human subjects tested, which are specific to epitopes present in Adalimumab, are not shared by Ab6, and thus not significantly recognized by Ab6. These results indicate potential use of the exemplified anti-human TNFα antibodies for treatment of patients who develop diminished clinical response or adverse reactions to anti-drug antibodies against other TNFα therapeutics such as Adalimumab.

Example 5: Immunogenicity Assessment

[0108] DC internalization assay, MAPPS assay, and T cell proliferation assay on LCDR1 and HCDR3 peptide clusters were performed to evaluate immunogenicity risk of the exemplified TNFα antibodies.

Dendritic cell internalization assay: The ability of human CD14+ monocytes derived from dendritic cells (DC) to internalize the exemplified anti-human TNFα antibodies was assessed. CD14+ monocytes were isolated from periphery blood mononuclear cells (PBMCs), cultured and differentiated into immature dendritic cells (with IL-4 and GM-CSF), all using standard protocols. To obtain mature DCs, cells were treated with 1 μg/mL LPS for 4 hours.

[0109] Exemplified anti-human TNFα antibodies were diluted at 8 μg/mL in complete RPMI medium and mixed at equal volume with detection probe Fab-TAMRA-QSY7 diluted to 5.33 m/mL in complete RPMI medium, and incubated for 30 min at 4° C. in the dark for complex formation, then added to immature and mature DC cultures and incubated for 24 h at 37° C. in a CO.sub.2 incubator. Cells were washed with 2% FBS PBS and resuspended in 100 μL 2% FBS PBS with Cytox Green live/dead dye. Data was collected on a BD LSR Fortessa X-20 and analyzed in FlowJo. Live single cells were gated, and percent of TAMRA fluorescence positive cells was recorded as the readout. To allow the comparison of molecules with data generated from different donors, a normalized internalization index was used. The internalization signal was normalized to IgG1 isotype (normalized internalization index=0) and an internal positive control PC (normalized internalization index=100) using Formula 1:

[00001]$\begin{array}{cc}100\times \frac{X_{T\mathrm{AMRA}}-I_{g}G1{\mathrm{isotype}}_{\mathrm{TAMRA}}}{P{C}_{TAMRA}-I_{g}G1{\mathrm{isotype}}_{TAMRA}}& \left(1\right)\end{array}$

where X.sub.TAMRA, IgG1 isotype.sub.TAMRA, and PC.sub.TAMRA were the percent of TAMRA-positive population for the test molecule X, IgG1 isotype, and PC respectively.

[0110] The results as demonstrated in Table 6, show that the tested anti-human TNFα antibodies were internalized into the cell upon binding to TNFα on the immature and mature dendritic cells.

TABLE-US-00007 TABLE 6 DC internalization of exemplified anti-human TNFα antibodies Immature Dendritic Cell Mature Dendritic Cell Antibody Internalization Index Internalization Index Ab1 19.4 26.7 Ab2 21.9 38.3 Ab3 20.0 35.2 Ab4 33.6 43.1 Ab5 54.6 52.7
MHC-associated peptide proteomics (MAPPs) Assay: MAPPs profiles the MHC-II presented peptides on human dendritic cells previously treated with exemplified anti-human TNFα antibodies. CD14+ monocytes were isolated from periphery blood mononuclear cells (PBMCs), cultured and differentiated into immature dendritic cells (with IL-4 and GM-CSF) using standard protocols. Exemplified antibodies were added to the immature dendritic cells on day 4 and fresh media containing LPS to transform the cells into mature dendritic cells was exchanged after 5-hour incubation. The matured dendritic cells were lysed in RIPA buffer with protease inhibitors and DNAse the following day. Immunoprecipitation of MHC-II complexes was performed using biotinylated anti-MHC-II antibody coupled to streptavidin beads. The bound complex was eluted and filtered. The isolated MHC-II peptides were analyzed by a mass spectrometer. Peptide identifications were generated by an internal proteomics pipeline using search algorithms with no enzyme search parameter against a bovine/human database with test sequences appended to the database. Peptides identified from the exemplified antibodies were aligned against the parent sequence.

[0111] The results as demonstrated in Table 7, showed that the exemplified anti-human TNFα antibodies had varying degree of presentation by MAPPs. Ab1 demonstrated the lowest MAPPs presentation with 1 non-germline cluster in 3 of the 10 donors tested.

TABLE-US-00008 TABLE 7 MAPPs analysis of exemplified anti-human TNFα antibodies Number of Number of non-germline donors containing Samples clusters across all donors ≥1 cluster Ab1 1 3/10 Ab2 2 6/10 Ab3 2 5/10 Ab4 3 5/10 Ab5 3 8/10
T cell proliferation assay: The ability of the exemplified anti-human TNFα antibodies MAPPs-derived peptide clusters to activate CD4+ T cells by inducing cellular proliferation was assessed. CD8+ T cells were depleted from cryopreserved PBMC's from 10 healthy donors and labeled with 1 μM Carboxyfluorescein Diacetate Succinimidyl Ester (CF SE). CD8+ T cell depleted PBMCs were seeded at 4×10.sup.6 cells/mL/well in AIM-V media (Life Technologies, cat #12055-083) with 5% CTS™ Immune Cell SR (Gibco, cat #A2596101) and tested in triplicate in 2.0 mL containing the different test molecules: DMSO control, media control, keyhole limpet haemocyanin (KLH; positive control), PADRE-X peptide (synthetic vaccine helper peptide, positive peptide control), or the respective anti-human TNFα antibody MAPPs-derived peptide clusters (10 μM each peptide). Cells were cultured and incubated for 7 days at 37° C. with 5% CO.sub.2. On day 7, samples were stained with the following cell surface markers: anti-CD3, anti-CD4, anti-CD14, anti-CD19, and DAPI for viability detection by flow cytometry using a BD LSRFortessa™, equipped with a High Throughput Sampler (HTS). Data was analyzed using FlowJo® Software (FlowJo, LLC, TreeStar) and a Cellular Division Index (CDI) was calculated. Briefly, the CDI for each MAPPs-derived peptide cluster was calculated by dividing the percent of proliferating CFSE.sup.dimCD4+ T cells from peptide-stimulated wells by the percent of proliferating CFSE.sup.dimCD4+ T cells in the unstimulated wells. A CDI of >2.5 was considered to represent a positive response. A percent donor frequency across all donors was evaluated.

[0112] The results as demonstrated in Tables 8a and 8b, show that the LCDR1 (Table 8a) and HCDR3 (Table 8b) peptides for Ab2 induced a T cell response frequency in about 22.0% and 25% donors respectively, indicating a significantly reduced immunogenicity risk for Ab2 when compared to the positive controls. The KLH positive control induced a T cell response in 100% of donors, and the PADRE-X (Synthetic vaccine helper peptide) positive control, induced a T cell response in 67% and 62.5% of donors respectively in the two studies. This range fell within the expected range for this assay (48.1%+24.4 Positive Donor Frequency).

TABLE-US-00009 TABLE 8a Frequency of CD4+ T cell responses induced by MAPPs-derived peptides in healthy donors. Median Median Number % CDI CDI of donors Molecule Positive (Positive (All Range positive Tested Donors Donors) donors) High Low (CDI > 2.5) KLH 100.0 190.8 190.8 1170.1 8.8 9/9 PADRE-X  67.0  4.0  3.1  17.8 0.5 6/9 Ab2 LCDR1  22.0  5.5  1.1  6.0 0.6 2/9 peptide

TABLE-US-00010 TABLE 8b Frequency of CD4+ T cell responses induced by MAPPs-derived peptides in healthy donors. Median Median % CDI CDI Number Molecule Positive (Positive (All Range of Tested Donors Donors) donors) High Low donors KLH 100.0 230.2 230.2 3558.5 12.0 8/8 PADRE-X  62.5  15.3   7.5   45.5  0.3 5/8 Ab2 HCDR3  25.0   7.2   1.3    7.7  0.1 2/8 peptide

Example 6. Biophysical Properties of Exemplified Anti-Human TNFα Antibodies

[0113] Biophysical properties of the exemplified anti-human TNFα antibodies were evaluated for developability.

Viscosity: Exemplified anti-human TNFα antibody samples were concentrated to about 125 mg/mL in a common formulation buffer matrix at pH 6. The viscosity for each antibody was measured using a VROC® initium (RheoSense) at 15° C. using the average of 9 replicate measurements. As demonstrated in Table 9, the results showed that the exemplified anti-human TNFα antibodies Ab1 (9.7 cP), Ab2 (9.2 cP), Ab3 (11.4 cP), and Ab4 (10.6 cP) had good viscosity profiles for developability.
Thermal stability: Differential Scanning calorimetry (DSC) was used to evaluate the stability of the exemplified antibodies against thermal denaturation. The thermal melting temperatures of the antibodies in PBS, pH 7.2 buffer, obtained by data fitting when unresolved, are listed in Table 9 (Tonset, TM1, TM2, and TM3). Although the thermal transitions for each domain were not all well resolved the data demonstrated in Table 9, show exemplified anti-human TNFα antibodies Ab1, Ab2, Ab3, and Ab4 had good thermal stability profiles for developability.
Aggregation upon temperature stress: The solution stability of the exemplified antibodies over time was assessed at approximately 100 mg/mL. Samples were incubated for a period of 28 days at 5° C. and 35° C. Following incubation, samples were analyzed for the percentage of high molecular weight (% HMW) species with size exclusion chromatography (SEC-HPLC). The results as demonstrated in Table 9, show exemplified anti-human TNFα antibodies Ab1, Ab2, Ab3, and Ab4 had good aggregation profiles for developability.

TABLE-US-00011 TABLE 9 Biophysical properties of exemplified anti-human TNFα antibodies % HMW % HMW Anti- Viscosity 28 d at 28 d at body Tonset TM1 TM2 TM3 (cP) 5° C. 35° C. Ab1 63.9 73.4 76.4 85.6  9.7 1.0 3.6 Ab2 64.8 72.5 81.0 87.4  9.2 0.6 4.3 Ab3 57.7 71.8 79.9 86.9 11.4 0.8 5.8 Ab4 64.9 73.1 79.7 87.2 10.6 0.7 3.3

Example 7: In Vivo Characterization of Exemplified Anti-Human TNFα Antibodies

[0114] Inhibition of human TNFα-induced CXCL1 cytokine production in vivo: Neutralization of TNFα-induced CXCL1 by the exemplified anti-human TNFα antibodies was assessed in vivo. Administration of human TNFα to C57/B6 mice induces a rapid and transient increase of mouse plasma CXCL1 levels. This allows for the interrogation of the neutralization capacity of the exemplified anti-human TNFα antibodies in vivo.

[0115] Briefly, C57/B6 mice (N=8/group) were subcutaneously administered with 0.3 mg/kg or 3 mg/kg of the exemplified antibodies or 3 mg/kg of a non-binding isotype control. Twenty-four hours post antibody administration, the mice were challenged by intraperitoneal injection of human TNFα at a dose of 3 μg/mouse. Two hours post human TNFα challenge the mice were sacrificed, blood was collected, and clarified to plasma by centrifugation. Plasma was analyzed for mouse CXCL1 concentration using a commercial MSD assay (MesoScale Discovery, P/N. K152QTG-1) according to manufacturer's instructions.

[0116] The results as demonstrated in Table 10, show that the exemplified anti-human TNFα antibodies significantly inhibited in vivo human TNFα-induced plasma CXCL1 production in a dose dependent manner, relative to isotype control treated mice (p<0.05, ANOVA followed by Turkey's Multiple Comparison test). Specifically, the exemplified anti-human TNFα antibodies inhibited TNFα induced in vivo plasma CXCL1 production by about 82% to about 93% at 3 mg/kg, and by about 46.5% to about 64.5% at 0.3 mg/kg. Thus, indicating that the exemplified anti-human TNFα antibodies neutralized the biological effects induced by human TNFα in vivo.

TABLE-US-00012 TABLE 10 Inhibition of human TNFα-induced CXCL1 cytokine production in vivo Plasma CXCL1 concentration Dose Mean ±SEM % P Value Antibody mg/kg (pg/mL) (pg/mL) Inhibition vs Isotype IgG1 isotype 3 1013.0 213.1  0.0% control Ab1 3 185.7 40.1 83.9% <0.0001 0.3 391.6 81.2 63.0% 0.0002 Ab2 3 203.4 33.3 82.1% <0.0001 0.3 554.4 59.3 46.5% 0.0164 Ab3 3 158.9 32.9 86.6% <0.0001 0.3 378.3 52.8 64.4% 0.0001 Ab4 3 100.1 10.3 92.6% <0.0001 0.3 449.3 52.8 57.2% 0.0009 Ab5 3 182.1 27.8 84.3% <0.0001 0.3 521.7 136.9 49.8% 0.0071 IgG1 isotype 3 27.3 9.2 100.0%  control without TNFα IP injection

TABLE-US-00013 SEQUENCE LISTING Ab1 SEQ ID NO: 1 HCDR1 for Ab1, Ab2, and Ab6 GYTFTGYYIH SEQ ID NO: 2 HCDR2 for Ab1, Ab2, and Ab6 WINPYTGGTNYAQKFQG SEQ ID NO: 3 HCDR3 for Ab1 DLYGSSNYGGDV SEQ ID NO: 4 LCDR1 for Ab1 and Ab3 QASQGISNYLN SEQ ID NO: 5 LCDR2 for Ab1, Ab2, Ab3, Ab4, Ab5, and Ab6 DASNLET SEQ ID NO: 6 LCDR3 for Ab1, Ab3, and Ab5 QQYDKLPLT SEQ ID NO: 7 VH for Ab1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYIHWVRQAPGQGLEWMGWIN PYTGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSNY GGDVWGQGTTVTVSS SEQ ID NO: 8 VL for Ab1 and Ab3 DIQMTQSPSSLSASVGDRVTITCQASQGISNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDKLPLTFGGGTKVEIK SEQ ID NO: 9 HC for Ab1 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYIHWVRQAPGQGLEWMGWIN PYTGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSNY GGDVWGQGTTVTVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD ICVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK SEQ ID NO: 10 LC for Ab1 and Ab3 DIQMTQSPSSLSASVGDRVTITCQASQGISNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDKLPLTFGGGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 11 HC DNA for Ab1 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG TGAAGGTCTCCTGCAAGGCTTCTGGATACACCTTCACCGGCTACTATATACAC TGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACC CTTACACCGGTGGCACAAACTATGCACAGAAGTTTCAGGGCAGGGTCACCAT GACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGCTGAGA TCTGACGACACGGCCGTGTATTACTGTGCGAGAGATCTCTATGGTTCGAGTAA TTACGGTGGCGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCT AGCACCAAGGGCCCATGTGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTG CCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAAC TCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAG GTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA ACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA GGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC TCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGA ACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAA GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCTGCGTGGA GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT GCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGA GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCT GCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO: 12 LC DNA for Ab1 and Ab3 GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAG AGTCACCATCACTTGCCAGGCGAGTCAGGGCATTAGCAACTATTTAAATTGGT ATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGATGCATCCAA TTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGAT TTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTG TCAACAGTATGATAAGCTCCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAG ATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGA GCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATC CCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAA CTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTC AGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA ACAGGGGAGAGTGC Ab2 SEQ ID NO: 1 HCDR1 for Ab1, Ab2, and Ab6 GYTFTGYYIH SEQ ID NO: 2 HCDR2 for Ab1, Ab2, and Ab6 WINPYTGGTNYAQKFQG SEQ ID NO: 13 HCDR3 for Ab2, Ab3, Ab4, and Ab5 DLYGSSNYGMDV SEQ ID NO: 14 LCDR1 for Ab2, Ab4, and Ab5 QASQGIRNYLN SEQ ID NO: 5 LCDR2 for Ab1, Ab2, Ab3, Ab4, Ab5, and Ab6 DASNLET SEQ ID NO: 15 LCDR3 for Ab2 and Ab4 QQYDNLPLT SEQ ID NO: 16 VH for Ab2 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYIHWVRQAPGQGLEWMGWIN PYTGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSNY GMDVWGQGTTVTVSS SEQ ID NO: 17 VL for Ab2 and Ab4 DIQMTQSPSSLSASVGDRVTITCQASQGIRNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDNLPLTFGGGTKVEIK SEQ ID NO: 18 HC for Ab2 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYIHWVRQAPGQGLEWMGWIN PYTGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSNY GMDVWGQGTTVTVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTV SWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKV DKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVS HEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEY KCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPS DICVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMH EALHNHYTQKSLSLSPGK SEQ ID NO: 19 LC for Ab2 and Ab4 DIQMTQSPSSLSASVGDRVTITCQASQGIRNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDNLPLTFGGGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 20 HC DNA for Ab2 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG TGAAGGTCTCCTGCAAGGCTTCTGGATACACCTTCACCGGCTACTATATACAC TGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACC CTTACACCGGTGGCACAAACTATGCACAGAAGTTTCAGGGCAGGGTCACCAT GACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGCTGAGA TCTGACGACACGGCCGTGTATTACTGTGCGAGAGATCTCTATGGTTCGAGTAA TTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCT AGCACCAAGGGCCCATGCGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTG CCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAAC TCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAG GTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA ACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA GGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC TCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGA ACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAA GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCTGCGTGGA GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT GCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGA GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCT GCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO: 21 LC DNA for Ab2 and Ab4 GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAG AGTCACCATCACTTGCCAGGCGAGTCAGGGCATTCGCAACTATTTAAATTGGT ATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGATGCATCCAA TTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGAT TTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTG TCAACAGTATGATAACCTCCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAG ATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGA GCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATC CCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAA CTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTC AGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA ACAGGGGAGAGTGC Ab3 SEQ ID NO: 22 HCDR1 for Ab3, Ab4, and Ab5 GYTFTGYYMH SEQ ID NO: 23 HCDR2 for Ab3, Ab4, and Ab5 WINPYTGGTKYAQKFQG SEQ ID NO: 13 HCDR3 for Ab2, Ab3, Ab4, and Ab5 DLYGSSNYGMDV SEQ ID NO: 4 LCDR1 for Ab2, Ab4, and Ab5 QASQGISNYLN SEQ ID NO: 5 LCDR2 for Ab1, Ab2, Ab3, Ab4, Ab5, and Ab6 DASNLET SEQ ID NO: 6 LCDR3 for Ab1, Ab3, and Ab5 QQYDKLPLT SEQ ID NO: 24 VH for Ab3, Ab4, and Ab5 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI NPYTGGTKYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSN YGMDVWGQGTTVTVSS SEQ ID NO: 8 VL for Ab1 and Ab3 DIQMTQSPSSLSASVGDRVTITCQASQGISNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDKLPLTFGGGTKVEIK SEQ ID NO: 25 HC for Ab3, Ab4, and Ab5 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI NPYTGGTKYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSN YGMDVWGQGTTVTVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK SEQ ID NO: 10 LC for Ab1 and Ab3 DIQMTQSPSSLSASVGDRVTITCQASQGISNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDKLPLTFGGGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 26 HC DNA for Ab3, Ab4, and Ab5 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG TGAAGGTCTCCTGCAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCAC TGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACC CTTACACCGGTGGCACAAAGTATGCACAGAAGTTTCAGGGCAGGGTCACCAT GACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGCTGAGA TCTGACGACACGGCCGTGTATTACTGTGCGAGAGATCTCTATGGTTCGAGTAA TTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCT AGCACCAAGGGCCCATGCGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTG CCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAAC TCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAG GTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA ACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA GGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC TCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGA ACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAA GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCTGCGTGGA GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT GCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGA GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCT GCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO: 12 LC DNA for Ab1 and Ab3 GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAG AGTCACCATCACTTGCCAGGCGAGTCAGGGCATTAGCAACTATTTAAATTGGT ATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGATGCATCCAA TTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGAT TTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTG TCAACAGTATGATAAGCTCCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAG ATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGA GCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATC CCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAA CTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTC AGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA ACAGGGGAGAGTGC Ab4 SEQ ID NO: 22 HCDR1 for Ab3, Ab4, and Ab5 GYTFTGYYMH SEQ ID NO: 23 HCDR2 for Ab3, Ab4, and Ab5 WINPYTGGTKYAQKFQG SEQ ID NO: 13 HCDR3 for Ab2, Ab3, Ab4, and Ab5 DLYGSSNYGMDV SEQ ID NO: 14 LCDR1 for Ab2, Ab4, and Ab5 QASQGIRNYLN SEQ ID NO: 5 LCDR2 for Ab1, Ab2, Ab3, Ab4, Ab5, and Ab6 DASNLET SEQ ID NO: 15 LCDR3 for Ab2 and Ab4 QQYDNLPLT SEQ ID NO: 24 VH for Ab3, Ab4, and Ab5 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI NPYTGGTKYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSN YGMDVWGQGTTVTVSS SEQ ID NO: 17 VL for Ab2 and Ab4 DIQMTQSPSSLSASVGDRVTITCQASQGIRNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDNLPLTFGGGTKVEIK SEQ ID NO: 25 HC for Ab3, Ab4, and Ab5 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI NPYTGGTKYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSN YGMDVWGQGTTVTVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK SEQ ID NO: 19 LC for Ab2 and Ab4 DIQMTQSPSSLSASVGDRVTITCQASQGIRNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDNLPLTFGGGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 26 HC DNA for Ab3, Ab4, and Ab5 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG TGAAGGTCTCCTGCAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCAC TGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACC CTTACACCGGTGGCACAAAGTATGCACAGAAGTTTCAGGGCAGGGTCACCAT GACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGCTGAGA TCTGACGACACGGCCGTGTATTACTGTGCGAGAGATCTCTATGGTTCGAGTAA TTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCT AGCACCAAGGGCCCATGCGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTG CCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAAC TCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAG GTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA ACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA GGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC TCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGA ACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAA GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCTGCGTGGA GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT GCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGA GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCT GCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO: 21 LC DNA for Ab2 and Ab4 GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAG AGTCACCATCACTTGCCAGGCGAGTCAGGGCATTCGCAACTATTTAAATTGGT ATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGATGCATCCAA TTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGAT TTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTG TCAACAGTATGATAACCTCCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAG ATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGA GCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATC CCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAA CTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTC AGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA ACAGGGGAGAGTGC Ab5 SEQ ID NO: 22 HCDR1 for Ab3, Ab4, and Ab5 GYTFTGYYMH SEQ ID NO: 23 HCDR2 for Ab3, Ab4, and Ab5 WINPYTGGTKYAQKFQG SEQ ID NO: 13 HCDR3 for Ab2, Ab3, Ab4, and Ab5 DLYGSSNYGMDV SEQ ID NO: 14 LCDR1 for Ab2, Ab4, and Ab5 QASQGIRNYLN SEQ ID NO: 5 LCDR2 for Ab1, Ab2, Ab3, Ab4, Ab5, and Ab6 DASNLET SEQ ID NO: 6 LCDR3 for Ab1, Ab3, and Ab5 QQYDKLPLT SEQ ID NO: 24 VH for Ab3, Ab4, and Ab5 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI NPYTGGTKYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSN YGMDVWGQGTTVTVSS SEQ ID NO: 27 VL for Ab5 DIQMTQSPSSLSASVGDRVTITCQASQGIRNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDKLPLTFGGGTKVEIK SEQ ID NO: 25 HC for Ab3, Ab4, and Ab5 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWI NPYTGGTKYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDLYGSSN YGMDVWGQGTTVTVSSASTKGPCVFPLAPSSKSTSGGTAALGCLVKDYFPEPVT VSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTK VDKRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDV SHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKE YKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFY PSDICVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSV MHEALHNHYTQKSLSLSPGK SEQ ID NO: 28 LC for Ab5 DIQMTQSPSSLSASVGDRVTITCQASQGIRNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDKLPLTFGGGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 26 HC DNA for Ab3, Ab4, and Ab5 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG TGAAGGTCTCCTGCAAGGCTTCTGGATACACCTTCACCGGCTACTATATGCAC TGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACC CTTACACCGGTGGCACAAAGTATGCACAGAAGTTTCAGGGCAGGGTCACCAT GACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGCTGAGA TCTGACGACACGGCCGTGTATTACTGTGCGAGAGATCTCTATGGTTCGAGTAA TTACGGTATGGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCT AGCACCAAGGGCCCATGCGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTG CCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAAC TCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAG GTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA ACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA GGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC TCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGA ACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAA GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCTGCGTGGA GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT GCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGA GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCT GCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO: 29 LC DNA for Ab5 GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAG AGTCACCATCACTTGCCAGGCGAGTCAGGGCATTCGCAACTATTTAAATTGGT ATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGATGCATCCAA TTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGAT TTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTG TCAACAGTATGATAAGCTCCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAG ATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGA GCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATC CCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAA CTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTC AGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA ACAGGGGAGAGTGC Ab6 SEQ ID NO: 1 HCDR1 for Ab1, Ab2, and Ab6 GYTFTGYYIH SEQ ID NO: 2 HCDR2 for Ab1, Ab2, and Ab6 WINPYTGGTNYAQKFQG SEQ ID NO: 30 HCDR3 for Ab6 DIYGSSNYGGDV SEQ ID NO: 31 LCDR1 for Ab6 QASQDISNYLN SEQ ID NO: 5 LCDR2 for Ab1, Ab2, Ab3, Ab4, Ab5, and Ab6 DASNLET SEQ ID NO: 32 LCDR3 for Ab6 QQYDTLPLT SEQ ID NO: 33 VH for Ab6 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYIHWVRQAPGQGLEWMGWIN PYTGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDIYGSSNY GGDVWGQGTTVTVSS SEQ ID NO: 34 VL for Ab6 DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDTLPLTFGGGTKVEIK SEQ ID NO: 35 HC for Ab6 QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYIHWVRQAPGQGLEWMGWIN PYTGGTNYAQKFQGRVTMTRDTSISTAYMELSRLRSDDTAVYYCARDIYGSSNY GGDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVS WNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVD KRVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSH EDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYK CKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSD IAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHE ALHNHYTQKSLSLSPGK SEQ ID NO: 36 LC for Ab6 DIQMTQSPSSLSASVGDRVTITCQASQDISNYLNWYQQKPGKAPKLLIYDASNLE TGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQYDTLPLTFGGGTKVEIKRTVAA PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQ ID NO: 37 HC DNA for Ab6 CAGGTGCAGCTGGTGCAGTCTGGGGCTGAGGTGAAGAAGCCTGGGGCCTCAG TGAAGGTCTCCTGCAAGGCTTCTGGATACACCTTCACCGGCTACTATATACAC TGGGTGCGACAGGCCCCTGGACAAGGGCTTGAGTGGATGGGATGGATCAACC CTTACACCGGTGGCACAAACTATGCACAGAAGTTTCAGGGCAGGGTCACCAT GACCAGGGACACGTCCATCAGCACAGCCTACATGGAGCTGAGCAGGCTGAGA TCTGACGACACGGCCGTGTATTACTGTGCGAGAGATATCTATGGTTCGAGTAA TTACGGTGGCGACGTCTGGGGCCAAGGGACCACGGTCACCGTCTCCTCAGCT AGCACCAAGGGCCCATCGGTCTTCCCCCTGGCACCCTCCTCCAAGAGCACCTC TGGGGGCACAGCGGCCCTGGGCTGCCTGGTCAAGGACTACTTCCCCGAACCG GTGACGGTGTCGTGGAACTCAGGCGCCCTGACCAGCGGCGTGCACACCTTCC CGGCTGTCCTACAGTCCTCAGGACTCTACTCCCTCAGCAGCGTGGTGACCGTG CCCTCCAGCAGCTTGGGCACCCAGACCTACATCTGCAACGTGAATCACAAGC CCAGCAACACCAAGGTGGACAAGAGAGTTGAGCCCAAATCTTGTGACAAAAC TCACACATGCCCACCGTGCCCAGCACCTGAACTCCTGGGGGGACCGTCAGTCT TCCTCTTCCCCCCAAAACCCAAGGACACCCTCATGATCTCCCGGACCCCTGAG GTCACATGCGTGGTGGTGGACGTGAGCCACGAAGACCCTGAGGTCAAGTTCA ACTGGTATGTGGACGGCGTGGAGGTGCATAATGCCAAGACAAAGCCGCGGGA GGAGCAGTACAACAGCACGTACCGTGTGGTCAGCGTCCTCACCGTCCTGCAC CAAGACTGGCTGAATGGCAAGGAGTACAAGTGCAAGGTCTCCAACAAAGCCC TCCCAGCCCCCATCGAGAAAACCATCTCCAAAGCCAAAGGGCAGCCCCGAGA ACCACAGGTGTACACCCTGCCCCCATCCCGGGAGGAGATGACCAAGAACCAA GTCAGCCTGACCTGCCTGGTCAAAGGCTTCTATCCCAGCGACATCGCCGTGGA GTGGGAGAGCAATGGGCAGCCGGAGAACAACTACAAGACCACGCCTCCCGT GCTGGACTCCGACGGCTCCTTCTTCCTCTATTCCAAGCTCACCGTGGACAAGA GCAGGTGGCAGCAGGGGAACGTCTTCTCATGCTCCGTGATGCATGAGGCTCT GCACAACCACTACACGCAGAAGAGCCTCTCCCTGTCTCCGGGCAAA SEQ ID NO: 38 LC DNA for Ab6 GACATCCAGATGACCCAGTCTCCATCCTCCCTGTCTGCATCTGTAGGAGACAG AGTCACCATCACTTGCCAGGCGAGTCAGGACATTAGCAACTATTTAAATTGGT ATCAGCAGAAACCAGGGAAAGCCCCTAAGCTCCTGATCTACGATGCATCCAA TTTGGAAACAGGGGTCCCATCAAGGTTCAGTGGAAGTGGATCTGGGACAGAT TTTACTTTCACCATCAGCAGCCTGCAGCCTGAAGATATTGCAACATATTACTG TCAACAGTATGATACCCTCCCGCTCACTTTCGGCGGAGGGACCAAGGTGGAG ATCAAACGGACCGTGGCTGCACCATCTGTCTTCATCTTCCCGCCATCTGATGA GCAGTTGAAATCTGGAACTGCCTCTGTTGTGTGCCTGCTGAATAACTTCTATC CCAGAGAGGCCAAAGTACAGTGGAAGGTGGATAACGCCCTCCAATCGGGTAA CTCCCAGGAGAGTGTCACAGAGCAGGACAGCAAGGACAGCACCTACAGCCTC AGCAGCACCCTGACGCTGAGCAAAGCAGACTACGAGAAACACAAAGTCTAC GCCTGCGAAGTCACCCATCAGGGCCTGAGCTCGCCCGTCACAAAGAGCTTCA ACAGGGGAGAGTGC SEQ ID NO: 39 Human TNFα protein MSTESMIRDVELAEEALPKKTGGPQGSRRCLFLSLFSFLIVAGATTLFCLLHFGVI GPQREEFPRDLSLISPLAQAVRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANA LLANGVELRDNQLVVPSEGLYLIYSQVLFKGQGCPSTHVLLTHTISRIAVSYQTKV NLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINRPDYLDFAES GQVYFGIIAL SEQ ID NO: 40 Rhesus macaque TNFα protein MSTESMIRDVELAEEALPRKTAGPQGSRRCWFLSLFSFLLVAGATTLFCLLHFGVI GPQREEFPKDPSLISPLAQAVRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRANA LLANGVELTDNQLVVPSEGLYLIYSQVLFKGQGCPSNHVLLTHTISRIAVSYQTK VNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINLPDYLDFAE SGQVYFGIIAL SEQ ID NO: 41 Mouse TNFα protein MSTESMIRDVELAEEALPQKMGGFQNSRRCLCLSLFSFLLVAGATTLFCLLNFGVI GPQRDEKFPNGLPLISSMAQTLTLRSSSQNSSDKPVAHVVANHQVEEQLEWLSQR ANALLANGMDLKDNQLVVPADGLYLVYSQVLFKGQGCPDYVLLTHTVSRFAIS YQEKVNLLSAVKSPCPKDTPEGAELKPWYEPIYLGGVFQLEKGDQLSAEVNLPK YLDFAESGQVYFGVIAL SEQ ID NO: 42 Rat TNFα protein MSTESMIRDVELAEEALPKKMGGLQNSRRCLCLSLFSFLLVAGATTLFCLLNFGV IGPNKEEKFPNGLPLISSMAQTLTLRSSSQNSSDKPVAHVVANHQAEEQLEWLSQ RANALLANGMDLKDNQLVVPADGLYLIYSQVLFKGQGCPDYVLLTHTVSRFAIS YQEKVSLLSAIKSPCPKDTPEGAELKPWYEPMYLGGVFQLEKGDLLSAEVNLPK YLDITESGQVYFGVIAL SEQ ID NO: 43 Rabbit TNFα protein MSTESMIRDVELAEGPLPKKAGGPQGSKRCLCLSLFSFLLVAGATTLFCLLHFRVI GPQEEESPNNLHLVNPVAQMVTLRSASRALSDKPLAHVVANPQVEGQLQWLSQ RANALLANGMKLTDNQLVVPADGLYLIYSQVLFSGQGCRSYVLLTHTVSRFAVS YPNKVNLLSAIKSPCHRETPEEAEPMAWYEPIYLGGVFQLEKGDRLSTEVNQPEY LDLAE SGQVYFGIIAL SEQ ID NO: 44 Canine TNFα protein MSTESMIRDVELAEEPLPKKAGGPPGSRRCFCLSLFSFLLVAGATTLFCLLHFGVI GPQREELPNGLQLISPLAQTVKSSSRTPSDKPVAHVVANPEAEGQLQWLSRRANA LLANGVELTDNQLIVPSDGLYLIYSQVLFKGQGCPSTHVLLTHTISRFAVSYQTKV NLLSAIKSPCQRETPEGTEAKPWYEPIYLGGVFQLEKGDRLSAEINLPNYLDFAES GQVYFGIIAL SEQ ID NO: 45 Cynomolgus monkey TNFα protein MSTESMIQDVELAEEALPRKTAGPQGSRRCWFLSLFSFLLVAGAATLFCLLHFGV IGPQREEFPKDPSLISPLAQAVRSSSRTPSDKPVAHVVANPQAEGQLQWLNRRAN ALVANGVELTDNQLVVPSEGLYLIYSQVLFKGQGCPSNHVLLTHTISRIAVSYQT KVNLLSAIKSPCQRETPEGAEAKPWYEPIYLGGVFQLEKGDRLSAEINLPDYLDFA ESGQVYFGIIAL SEQ ID NO: 46 LCDR1 consensus sequence QASQGIXaa.sub.7NYLN Wherein Xaa.sub.7 is Serine or Arginine SEQ ID NO: 47 LCDR3 consensus sequence QQYDXaa.sub.5LPLT Wherein Xaa5 is Asparagine or Lysine SEQ ID NO: 48 human TNFR1 MGLSTVPDLLLPLVLLELLVGIYPSGVIGLVPHLGDREKRDSVCPQGKYIHPQNNS ICCTKCHKGTYLYNDCPGPGQDTDCRECESGSFTASENHLRHCLSCSKCRKEMG QVEISSCTVDRDTVCGCRKNQYRHYWSENLFQCFNCSLCLNGTVHLSCQEKQNT VCTCHAGFFLRENECVSCSNCKKSLECTKLCLPQIENVKGTEDSGTTVLLPLVIFF GLCLLSLLFIGLMYRYQRWKSKLYSIVCGKSTPEKEGELEGTTTKPLAPNPSFSPT PGFTPTLGFSPVPSSTFTSSSTYTPGDCPNFAAPRREVAPPYQGADPILATALASDPI PNPLQKWEDSAHKPQSLDTDDPATLYAVVENVPPLRWKEFVRRLGLSDHEIDRL ELQNGRCLREAQYSMLATWRRRTPRREATLELLGRVLRDMDLLGCLEDIEEALC GPAALPPAPSLLR SEQ ID NO: 49 human TNFR2 MAPVAVWAALAVGLELWAAAHALPAQVAFTPYAPEPGSTCRLREYYDQTAQM CCSKCSPGQHAKVFCTKTSDTVCDSCEDSTYTQLWNWVPECLSCGSRCSSDQVE TQACTREQNRICTCRPGWYCALSKQEGCRLCAPLRKCRPGFGVARPGTETSDVV CKPCAPGTFSNTTSSTDICRPHQICNVVAIPGNASMDAVCTSTSPTRSMAPGAVHL PQPVSTRSQHTQPTPEPSTAPSTSFLLPMGPSPPAEGSTGDFALPVGLIVGVTALGL LIIGVVNCVIMTQVKKKPLCLQREAKVPHLPADKARGTQGPEQQHLLITAPSSSSS SLESSASALDRRAPTRNQPQAPGVEASGAGEARASTGSSDSSPGGHGTQVNVTCI VNVCSSSDHSSQCSSQASSTMGDTDSSPSESPKDEQVPFSKEECAFRSQLETPETL LGSTEEKPLPLGVPDAGMKPS