CGRP ANTIBODIES AND USES THEREOF

Abstract

The present invention relates to antibodies that bind to human CGRP, compositions and kits comprising such CGRP antibodies, and methods of using such CGRP antibodies for detection of human CGRP.

Claims

1. An antibody or antibody fragment thereof that binds to human CGRP and comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises the amino acid sequence of SEQ ID NO: 1 at CDRL1, the amino acid sequence of SEQ ID NO: 2 at CDRL2, and the amino acid sequence of SEQ ID NO: 3 at CDRL3, and wherein the HCVR comprises the amino acid sequence of SEQ ID NO: 4 at CDRH1, the amino acid sequence of SEQ ID NO: 5 at CDRH2, and the amino acid sequence of SEQ ID NO: 6 at CDRH3.

2. The antibody or antibody fragment of claim 1, wherein the LCVR has the amino acid sequence of SEQ ID NO:7 and the HCVR has the amino acid sequence of SEQ ID NO: 8.

3. The antibody of claim 1, further comprising a light chain (LC) and a heavy chain (HC), wherein the LC has the amino acid sequence of SEQ ID NO: 9 and the HC has the amino acid sequence of SEQ ID NO: 10.

4. The antibody of claim 3, wherein the antibody comprises two LCs and two HCs, wherein each LC has the amino acid sequence of SEQ ID NO: 9, and each HC has the amino acid sequence of SEQ ID NO: 10.

5. The antibody fragment of claim 1, wherein the antibody fragment is an antibody Fab fragment.

6. The antibody or antibody fragment according to claim 1, wherein the antibody or antibody fragment is labelled with a detectable label.

7. A kit comprising a first CGRP antibody comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises the amino acid sequence of SEQ ID NO: 1 at CDRL1, the amino acid sequence of SEQ ID NO: 2 at CDRL2, and the amino acid sequence of SEQ ID NO: 3 at CDRL3, and wherein the HCVR comprises the amino acid sequence of SEQ ID NO: 4 at CDRH1, the amino acid sequence of SEQ ID NO: 5 at CDRH2, and the amino acid sequence of SEQ ID NO: 6 at CDRH3 and a second CGRP antibody.

8. (canceled)

9. The kit of claim 7, wherein the second CGRP antibody comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR) wherein the LCVR comprises the amino acid sequence of SEQ ID NO: 13 at CDRL1, the amino acid sequence of SEQ ID NO: 14 at CDRL2, and the amino acid sequence of SEQ ID NO: 15 at CDRL3, and wherein the HCVR comprises the amino acid sequence of SEQ ID NO: 16 at CDRH1, the amino acid sequence of SEQ ID NO: 17 at CDRH2, and the amino acid sequence of SEQ ID NO: 18 at CDRH3.

10. The kit of claim 7, wherein the second CGRP antibody comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR) wherein the LCVR comprises the amino acid sequence of SEQ ID NO: 19 at CDRL1, the amino acid sequence of SEQ ID NO: 20 at CDRL2, and the amino acid sequence of SEQ ID NO: 21 at CDRL3, and wherein the HCVR comprises the amino acid sequence of SEQ ID NO: 22 at CDRH1, the amino acid sequence of SEQ ID NO: 23 at CDRH2, and the amino acid sequence of SEQ ID NO: 24 at CDRH3.

11. A method of detecting human CGRP in a patient sample comprising contacting the patient sample with a first CGRP antibody, and second CGRP antibody comprising a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises the amino acid sequence of SEQ ID NO: 1 at CDRL1, the amino acid sequence of SEQ ID NO: 2 at CDRL2, and the amino acid sequence of SEQ ID NO: 3 at CDRL3, and wherein the HCVR comprises the amino acid sequence of SEQ ID NO: 4 at CDRH1, the amino acid sequence of SEQ ID NO: 5 at CDRH2, and the amino acid sequence of SEQ ID NO: 6 at CDRH3 and measuring the amount of the first or second antibody that is bound to the human CGRP in order to determine the amount of human CGRP in the patient sample.

12. The method of claim 11, wherein the patient sample comprises plasma, EDTA plasma, heparin plasma, serum, CSF, or synovial fluid.

13. (canceled)

14. The method of claim 11, wherein the first CGRP antibody comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR) wherein the LCVR comprises the amino acid sequence of SEQ ID NO: 13 at CDRL1, the amino acid sequence of SEQ ID NO: 14 at CDRL2, and the amino acid sequence of SEQ ID NO: 15 at CDRL3, and wherein the HCVR comprises the amino acid sequence of SEQ ID NO: 16 at CDRH1, the amino acid sequence of SEQ ID NO: 17 at CDRH2, and the amino acid sequence of SEQ ID NO: 18 at CDRH3.

15. The method of claim 11, wherein the first CGRP antibody comprises a light chain variable region (LCVR) and a heavy chain variable region (HCVR), wherein the LCVR comprises the amino acid sequence of SEQ ID NO: 19 at CDRL1, the amino acid sequence of SEQ ID NO: 20 at CDRL2, and the amino acid sequence of SEQ ID NO: 21 at CDRL3, and wherein the HCVR comprises the amino acid sequence of SEQ ID NO: 22 at CDRH1, the amino acid sequence of SEQ ID NO: 23 at CDRH2, and the amino acid sequence of SEQ ID NO: 24 at CDRH3.

16. The method of claim 11, wherein the first CGRP antibody is labeled with a detectable label.

17. The method of claim 11, wherein the second CGRP antibody is labeled with a detectable label.

18. (canceled)

19. (canceled)

20. (canceled)

21. (canceled)

22. (canceled)

23. (canceled)

24. (canceled)

25. (canceled)

26. (canceled)

27. (canceled)

28. (canceled)

Description

EXAMPLES

Antibody Expression and Purification

[0020] The CGRP-reactive antibody CA1 1 is expressed transiently in HEK293 cells.

[0021] The antibody is mouse IgGl/kappa and was purified using protein G. Briefly, 2 L of HEK293 supernatant from cells transfected with LC and HC vectors of mIgGl CA11 are harvested five days post transfection and loaded at 2 mL/min overnight in cold room onto a 5 mL, Protein G Column (GE Healthcare #17-0405-03). The next day, Protein G column is washed at 5 mL/min with 5 column volumes of PBS, pH 7.4. The bound CA11 antibody is eluted from Protein G column at 5 mL/min with 10 mM citric acid, pH 3, and immediately neutralized with 1/10 volume of 1 M Tris, pH 8. The CA11 antibody is buffer exchanged into PBS, pH 7.4, and concentrated in Millipore centrifugal concentrators to 1.7 mg/mL. Purity of the antibody is assessed using SDS-PAGE and size-exclusion chromatography. N-terminal sequencing and MALDI-TOF are used to further confirm the identity of the CA11 antibody. BIAcore binding is performed to determine binding affinity of CA11 antibody to CGRP peptide. Sequences of the exemplified antibodies are provided in Table 1.

TABLE-US-00001 TABLE 1 SEQ IDs of amino acid sequences of the exemplified antibodies. Light Heavy Antibody Chain Chain LCVR HCVR CA11 9 10 7 8 Antibody I 25 26 Antibody II 27 28 Antibody LCDR1 LCDR2 LCDR3 CA11 1 2 3 Antibody I 13 14 15 Antibody II 19 20 21 Antibody HCDR1 HCDR2 HCDR3 CA11 4 5 6 Antibody I 16 17 18 Antibody II 22 23 24

CGRP Fab Binding ELISA

[0022] An ELISA based assay is used to measure the relative affinities of CA11 and parent (C1WT) Fab molecules. Briefly, 96 well plates are coated by adding 50 L/well of a 1 g/mL solution containing goat anti-human kappa (Southern Biotech #2060-01) in PBS 7.4 overnight at 4 C. Following incubation, plates are blocked with 200 L/well of a Casein/PBS solution (Thermo-Fisher Scientific #37528) for 1 hr at room temperature. Plates are washed 3 times with PBS containing 0.1% Tween20 (PBS-T). Fifty L of each Fab are normalized to a concentration of 2 g/mL, added to separate columns on the plate, and incubated for 1 hr at 37 C. The plate is washed 3 times with PBS-T prior to incubation with 50 L of N-terminal biotin labelled human CGRP (Abgent Custom Synthesized Peptide #355061532). Starting at 20 nM, three-fold serial dilutions of the N-terminal biotin labelled CGRP peptide are added to the captured Fabs and incubated for 1 hr at 37 C. The plate is washed 3 times with PBS-T. To facilitate dissociation of the human CGRP peptide from the captured Fabs, an additional 200 L of PBS-T is added to each well and incubated for 2 hr at 37 C. The plate is washed, 50 L of alkaline phosphatase conjugated neutravidin (Pierce #31002) diluted 1:1000 in PBS-T is added, and the plates are incubated for 1 hr at 37 C. The plates are washed and 50 L of alkaline phosphatase substrate diluted 25-fold in water is added. Following colorimetric development, the plate is read using a Molecular Devices VMax Kinetic ELISA Microplate Reader and the absorbance at 560 nm is recorded as a function of human CGRP concentration, plotting with Microsoft Excel.

TABLE-US-00002 TABLE 2 ELISA binding data. CGRP (nM) C1WT (A560) CA11 (A560) 20 0.327 1.046 6.667 0.232 1.028 2.222 0.14 0.688 0.741 0.084 0.356 0.247 0.064 0.165 0.082 0.051 0.091 0.027 0.052 0.063 0.009 0.065 0.082

[0023] The data shown in Table 2 demonstrate that CA11 binds CGRP.

High Sensitive CGRP MSD (Meso Scale Discovery) ELISA

[0024] A Meso Scale Discovery (MSD) assay is used to determine the ability of CA11 antibody in detecting human CGRP. Plates are blocked by adding 150 L/well 3% Blocker A/PBS, and incubating for 60 minutes at room temperature with rotation at 650 rpm. Plates are washed 3 times with PBS-T, and diluted biotin-labelled anti-CGRP antibody (Antibody I; 0.1 g/mL in 0.1% Blocker A/PBS) is added into wells of streptavidin plates. Plates are incubated for 1 hour at room temperature with rotation (650 rpm).

[0025] Plates are washed, 25 1 of human healthy donor serum, heparin plasma, CSF, or synovial fluid samples (or standard; alpha-CGRP; Bachem) are added into wells, and incubated at room temperature with rotation for 2 hours. Plates are washed, and 25 L Sulfo-Tag labelled-anti-CGRP (CA11) (0.5 g/mL) is added, and plates are incubated for 1 hour at room temperature with rotation. Plates are washed and 150 uL/ well 2 MSD Read Buffer T is added to each well. Plates are read on MSD instrument, and unknowns are calculated using a log-log 4-5 PL fit on the MSD Discovery Workbench software or equivalent. The CGRP concentration from human donor samples is summarized in Table 2.

[0026] To determine the spike and recovery in each of matrices, the different concentration of CGRP standard is spiked into human EDTA plasma, heparin plasma, serum, CSF or synovial fluid. The recovery of CGRP in each of matrices is summarized in Table 3.

TABLE-US-00003 TABLE 2 Summary of CGRP level in healthy donors. CGRP Concentration Matrix Sample Number (pg/mL) Mean +/ SD EDTA Plasma 55 2.2 +/ 0.86 Serum 61 1.78 +/ 0.60 Heparin Plasma 10 1.23 +/ 1.03 CSF 20 5.48 +/ 1.61 Synovial Fluid 6 0.32 +/ 0.25

[0027] The data in Table 2 demonstrate that the CGRP MSD assay with the CA11 antibody detected CGRP in healthy donors.

TABLE-US-00004 TABLE 3 Spike and recovery of CGRP. Spike % Spike % (pg/mL) Recovery (pg/mL) Recovery EDTA 25 98 CSF 25 107 Plasma 8.33 98 8.33 103 2.78 99 2.78 99 0.93 97 0.93 98 0.31 101 0.31 99 Serum 25 83 Synovial 25 103 8.33 84 Fluid 8.33 99 2.78 88 2.78 92 0.93 87 0.93 91 0.31 96 0.31 90 Heparin 30 107 Plasma 3.3 112 0.37 115 0.12 108

[0028] The data in Table 3 demonstrate that the CGRP MSD assay with the CA11 antibody detected CGRP in human EDTA plasma, serum, heparin, CSF, and synovial fluid.

Quanterix Simoa Assay

[0029] Beads (0.5 mg/ml) are conjugated to Antibody II according to the Quanterix protocol. A 10 ml solution of beads (5 million beads/mL), a 10 mL solution of biotinylated CA11 antibody (0.1 g/mL), and a 10 mL solution of streptavidin-beta-galactosidase (SBG; 150 pM) are prepared and transferred to separate 15 mL bottles. Beads, CA11 antibody, calibrators, SBG, and supplied resorufin-beta-D galactopyranoside RGP reagents are loaded into the instrument according to the Simoa HD-1 Analyzer User Guide. The run is initiated and run on the instrument according to the Homebrew chapter of the Simoa HD-1 Analyzer User Guide. Binding data is shown in Table 4.

[0030] To determine the spike and recovery in the Quanterix assay with the CA11 antibody, CGRP is spiked into the human plasma. The percentage of recovered spiked CGRP is summarized in Table 5.

[0031] To determine the sensitivity of the CGRP Quanterix assay with the CA11 antibody, CGRP levels in healthy donor plasma are detected. The concentration of CGRP detected from healthy donor plasma is shown in Table 6.

TABLE-US-00005 TABLE 4 CGRP Quanterix Assay Binding Data. CGRP (pg/mL) Average AEB* 50 2.99 16.67 1.18 5.56 0.42 1.85 0.13 0.62 0.06 0.21 0.02 0.07 0.011 0.02 0.0075 0.0076 0.0079 0 0.0064 *AEB = average enzyme per bead

[0032] The data in Table 4 demonstrate that the CGRP Quanterix assay with the CA11 antibody detected human CGRP in plasma as low as 0.02 pg/ml.

TABLE-US-00006 TABLE 5 CGRP spike and recovery in EDTA plasma. Spike (pg/mL) Recovery (%) 25 83 8.33 77 2.78 78 0.93 77 0.31 99 0.10 96

[0033] The data in Table 5 demonstrate that the CGRP Quanterix assay with the CA11 antibody detected spiked CGRP in EDTA plasma with a percent recovery in the range of 77% to 99%.

TABLE-US-00007 TABLE 6 Plasma CGRP level in healthy donors (n = 9 donors per group). CGRP Concentration Matrix (pg/mL) Mean +/ SD EDTA Plasma 0.93 +/ 0.64 Heparin Plasma 1.02 +/ 0.77

[0034] The data in Table 6 demonstrate that the CGRP Quanterix assay with the CA11 antibody detected about 0.93 pg/mL CGRP in healthy donor EDTA plasma, and about 1.02 pg/mL CGRP in healthy donor heparin plasma.

TABLE-US-00008 Sequences SEQIDNO:1(CA11LCDR1) SASSSISSIYLH SEQIDNO:2(CA11LCDR2) YRAKNLAS SEQIDNO:3(CA11LCDR3) QQGSTIPFT SEQIDNO:4(CA11HCDR1) KASGYTFTRSVMH SEQIDNO:5(CA11HCDR2) YINPYNDGTKYNEKFKG SEQIDNO:6(CA11HCDR3) AKSGNDGY SEQIDNO:7(CA11LCVR) EIVLTQSPTTMAASPGEKITITCSASSSISSIYLHWYQQKPGFSPKVLIYRAKNLASG VPARFSGSGSGTSYSLTIGTMEAEDVATYYCQQGSTIPFTFGSGTKLEIK SEQIDNO:8(CA11HCVR) EVQLQQSGPELVKPGASVKMSCKASGYTFTRSVMHWVKQKPGQGLEWIGYINP YNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCAKSGNDGYWG QGTTLTVSS SEQIDNO:9(CA11LC) EIVLTQSPTTMAASPGEKITITCSASSSISSIYLHWYQQKPGFSPKVLIYRAKNLASG VPARFSGSGSGTSYSLTIGTMEAEDVATYYCQQGSTIPFTFGSGTKLEIKRADAAP TVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDS KDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC SEQIDNO:10(CA11HC) EVQLQQSGPELVKPGASVKMSCKASGYTFTRSVMHWVKQKPGQGLEWIGYINP YNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCAKSGNDGYWG QGTTLTVSSAKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSL SSGVHTFPAVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDC GCKPCICTVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDV EVHTAQTQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISK TKGRPKAPQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYK NTQPIMDTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPG K SEQIDNO:11(CA11LCDNA) GAAATCGTGCTGACCCAGAGCCCCACCACCATGGCCGCCAGCCCTGGCGAGA AGATCACCATCACCTGCTCCGCCAGCAGCAGCATCAGCTCCATCTACCTGCAC TGGTATCAGCAGAAGCCCGGCTTCAGCCCTAAGGTGCTGATCTACCGGGCCA AAAACCTGGCCAGCGGCGTGCCCGCCAGATTCAGCGGCAGCGGCTCCGGCAC CAGCTACAGCCTGACCATCGGCACCATGGAGGCCGAGGACGTGGCCACCTAC TACTGCCAGCAGGGCAGCACCATCCCCTTCACCTTCGGCAGCGGCACCAAGC TGGAGATCAAGCGGGCTGATGCGGCGCCCACTGTATCCATCTTCCCACCATCC AGTGAGCAGTTAACATCTGGAGGTGCCTCAGTCGTGTGCTTCTTGAACAACTT CTACCCCAAAGACATCAATGTCAAGTGGAAGATTGATGGCAGTGAACGACAA AATGGCGTCCTGAACAGTTGGACTGATCAGGACAGCAAAGACAGCACCTACA GCATGAGCAGCACCCTCACGTTGACCAAGGACGAGTATGAACGACATAACAG CTATACCTGTGAGGCCACTCACAAGACATCAACTTCACCCATTGTCAAGAGCT TCAACAGGAATGAGTGT SEQIDNO:12(CA11HCDNA) GAAGTGCAGCTGCAGCAGAGCGGCCCTGAGCTGGTGAAGCCTGGCGCCAGCG TGAAGATGAGCTGTAAGGCCAGCGGCTACACCTTCACCAGGAGCGTGATGCA CTGGGTGAAGCAGAAGCCCGGCCAGGGCCTGGAGTGGATCGGCTACATCAAC CCCTACAACGACGGCACCAAGTACAACGAGAAGTTCAAGGGCAAGGCCACCC TGACCAGCGACAAGAGCAGCAGCACCGCCTACATGGAGCTGTCCAGCCTGAC AAGCGAGGATAGCGCCGTGTACTACTGTGCCAAGTCGGGCAATGACGGCTAC TGGGGCCAGGGCACCACACTGACCGTGTCCAGCGCCAAAACGACACCCCCAT CTGTCTATCCGCTAGCCCCTGGATCTGCCGCCCAGACCAACAGCATGGTGACC CTGGGCTGTCTGGTGAAGGGCTACTTCCCTGAGCCTGTGACAGTGACCTGGAA CAGCGGCTCTCTGTCTAGCGGCGTGCACACATTCCCTGCCGTGCTGCAGAGCG ACCTGTACACCCTGAGCAGCAGCGTGACCGTGCCTAGCAGCACATGGCCTAG CGAGACCGTGACATGCAACGTGGCCCACCCTGCCTCTTCTACCAAGGTGGAC AAGAAGATCGTGCCCAGAGACTGCGGCTGCAAGCCTTGCATCTGCACCGTGC CTGAGGTGAGCAGCGTGTTCATCTTCCCACCCAAGCCCAAGGACGTGCTCACC ATCACCCTCACCCCCAAGGTCACGTGTGTTGTGGTAGACATCAGCAAGGATG ATCCCGAGGTCCAGTTCAGCTGGTTTGTAGATGATGTGGAGGTGCACACAGCT CAGACGCAACCCCGGGAGGAGCAGTTCAACAGCACTTTCCGCTCAGTCAGTG AACTTCCCATCATGCACCAGGACTGGCTCAATGGCAAGGAGTTCAAATGCAG GGTCAACAGTGCAGCTTTCCCTGCCCCCATCGAGAAAACCATCTCCAAAACC AAAGGCAGACCGAAGGCTCCACAGGTGTACACCATTCCACCTCCCAAGGAGC AGATGGCCAAGGATAAAGTCAGTCTGACCTGCATGATAACAGACTTCTTCCCT GAAGACATTACTGTGGAGTGGCAGTGGAATGGGCAGCCAGCGGAGAACTAC AAGAACACTCAGCCCATCATGGACACAGATGGCTCTTACTTCGTCTACAGCA AGCTCAATGTGCAGAAGAGCAACTGGGAGGCAGGAAATACTTTCACCTGCTC TGTGTTACATGAGGGCCTGCACAACCACCATACTGAGAAGAGCCTCTCCCACT CTCCTGGTAAA SEQIDNO:13(AntibodyILCDR1) RASQDIDNYLN SEQIDNO:14(AntibodyILCDR2) YTSEYHS SEQIDNO:15(AntibodyILCDR3) QQGDALPPT SEQIDNO:16(AntibodyIHCDR1) GYTFGNYWMQ SEQIDNO:17(AntibodyIHCDR2) AIYEGTGDTRYIQKFAG SEQIDNO:18(AntibodyIHCDR3) LSDYVSGFSY SEQIDNO:19(AntibodyIILCDR1) RASKDISKYLN SEQIDNO:20(AntibodyIILCDR2) YTSGYHS SEQIDNO:21(AntibodyIILCDR3) QQGDALPPT SEQIDNO:22(AntibodyIIHCDR1) GYTFGNYWMQ SEQIDNO:23(AntibodyIIHCDR2) AIYEGTGKTVYIQKFAD SEQIDNO:24(AntibodyIIHCDR3) LSDYVSGFGY SEQIDNO:25(AntibodyILC) DIQMTQSPSSLSASVGDRVTITCRASQDIDNYLNWYQQKPGKAPKLLIYYTSEYH SGVPSRFSGSGSGTDFTFTISSLQPEDIATYYCQQGDALPPTFGQGTKLEIKRTVAA PSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQD SKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQIDNO:26(AntibodyIHC) QVQLVQSGAEVKKPGASVKVSCKASGYTFGNYWMQWVRQAPGQGLEWMGAI YEGTGDTRYIQKFAGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARLSDYVSG FSYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSW NSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDK RVESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPE VQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVS NKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHN HYTQKSLSLSLG SEQIDNO:27(AntibodyIILC) DIQMTQSPSSLSASVGDRVTITCRASKDISKYLNWYQQKPGKAPKLLIYYTSGYH SGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQGDALPPTFGGGTKVEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQ DSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC SEQIDNO:28(AntibodyIIHC) QVQLVQSGAEVKKPGSSVKVSCKASGYTFGNYWMQWVRQAPGQGLEWMGAI YEGTGKTVYIQKFADRVTITADKSTSTAYMELSSLRSEDTAVYYCARLSDYVSGF GYWGQGTTVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWN SGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKR VESKYGPPCPPCPAPEAAGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEV QFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEW ESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNH YTQKSLSLSLG