TW201111394A - Bispecific, tetravalent antigen binding proteins - Google Patents
Bispecific, tetravalent antigen binding proteins Download PDFInfo
- Publication number
- TW201111394A TW201111394A TW099119745A TW99119745A TW201111394A TW 201111394 A TW201111394 A TW 201111394A TW 099119745 A TW099119745 A TW 099119745A TW 99119745 A TW99119745 A TW 99119745A TW 201111394 A TW201111394 A TW 201111394A
- Authority
- TW
- Taiwan
- Prior art keywords
- antibody
- domain
- ser
- val
- antigen
- Prior art date
Links
- 102000025171 antigen binding proteins Human genes 0.000 title claims abstract description 75
- 108091000831 antigen binding proteins Proteins 0.000 title claims abstract description 75
- 238000000034 method Methods 0.000 claims abstract description 57
- 239000008194 pharmaceutical composition Substances 0.000 claims abstract description 11
- 230000027455 binding Effects 0.000 claims description 78
- 239000000427 antigen Substances 0.000 claims description 70
- 108091007433 antigens Proteins 0.000 claims description 70
- 102000036639 antigens Human genes 0.000 claims description 70
- 235000001014 amino acid Nutrition 0.000 claims description 17
- 206010028980 Neoplasm Diseases 0.000 claims description 16
- 125000000539 amino acid group Chemical group 0.000 claims description 14
- 150000001413 amino acids Chemical class 0.000 claims description 14
- 230000015572 biosynthetic process Effects 0.000 claims description 12
- 201000011510 cancer Diseases 0.000 claims description 12
- 239000013598 vector Substances 0.000 claims description 12
- 238000011282 treatment Methods 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 9
- 239000002253 acid Substances 0.000 claims description 7
- 150000001412 amines Chemical class 0.000 claims description 7
- 238000003786 synthesis reaction Methods 0.000 claims description 7
- 230000008859 change Effects 0.000 claims description 4
- 239000003814 drug Substances 0.000 claims description 4
- 239000004475 Arginine Substances 0.000 claims description 3
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims description 3
- MTCFGRXMJLQNBG-UHFFFAOYSA-N Serine Natural products OCC(N)C(O)=O MTCFGRXMJLQNBG-UHFFFAOYSA-N 0.000 claims description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 claims description 3
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 2
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 claims description 2
- ONIBWKKTOPOVIA-UHFFFAOYSA-N Proline Natural products OC(=O)C1CCCN1 ONIBWKKTOPOVIA-UHFFFAOYSA-N 0.000 claims description 2
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 claims description 2
- 235000004279 alanine Nutrition 0.000 claims description 2
- 235000018417 cysteine Nutrition 0.000 claims description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims description 2
- 241000282320 Panthera leo Species 0.000 claims 1
- 239000011148 porous material Substances 0.000 claims 1
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 210000004027 cell Anatomy 0.000 description 83
- 108090000623 proteins and genes Proteins 0.000 description 54
- 102000004169 proteins and genes Human genes 0.000 description 44
- 235000018102 proteins Nutrition 0.000 description 43
- 230000035772 mutation Effects 0.000 description 38
- 102000009524 Vascular Endothelial Growth Factor A Human genes 0.000 description 30
- 108010019530 Vascular Endothelial Growth Factors Proteins 0.000 description 27
- 108090000765 processed proteins & peptides Proteins 0.000 description 25
- 239000000203 mixture Substances 0.000 description 21
- 239000000872 buffer Substances 0.000 description 20
- 108020004414 DNA Proteins 0.000 description 17
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 17
- 229920001213 Polysorbate 20 Polymers 0.000 description 16
- 230000004927 fusion Effects 0.000 description 16
- 210000002706 plastid Anatomy 0.000 description 16
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 16
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 16
- 150000007523 nucleic acids Chemical class 0.000 description 15
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 14
- 229940024606 amino acid Drugs 0.000 description 14
- 102000039446 nucleic acids Human genes 0.000 description 13
- 108020004707 nucleic acids Proteins 0.000 description 13
- 102000004196 processed proteins & peptides Human genes 0.000 description 12
- 108060003951 Immunoglobulin Proteins 0.000 description 11
- 230000014509 gene expression Effects 0.000 description 11
- 102000018358 immunoglobulin Human genes 0.000 description 11
- 241000880493 Leptailurus serval Species 0.000 description 10
- 238000004113 cell culture Methods 0.000 description 10
- 239000012634 fragment Substances 0.000 description 10
- 229920001184 polypeptide Polymers 0.000 description 10
- 101100481408 Danio rerio tie2 gene Proteins 0.000 description 9
- 101100481410 Mus musculus Tek gene Proteins 0.000 description 9
- 238000004458 analytical method Methods 0.000 description 9
- 238000001514 detection method Methods 0.000 description 9
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 8
- 125000003275 alpha amino acid group Chemical group 0.000 description 8
- 239000006227 byproduct Substances 0.000 description 8
- 238000010168 coupling process Methods 0.000 description 8
- 239000012228 culture supernatant Substances 0.000 description 8
- 239000012636 effector Substances 0.000 description 8
- 150000002482 oligosaccharides Polymers 0.000 description 8
- HKZAAJSTFUZYTO-LURJTMIESA-N (2s)-2-[[2-[[2-[[2-[(2-aminoacetyl)amino]acetyl]amino]acetyl]amino]acetyl]amino]-3-hydroxypropanoic acid Chemical compound NCC(=O)NCC(=O)NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O HKZAAJSTFUZYTO-LURJTMIESA-N 0.000 description 7
- 238000002965 ELISA Methods 0.000 description 7
- 210000004899 c-terminal region Anatomy 0.000 description 7
- 230000004540 complement-dependent cytotoxicity Effects 0.000 description 7
- 230000008878 coupling Effects 0.000 description 7
- 238000005859 coupling reaction Methods 0.000 description 7
- 229920001542 oligosaccharide Polymers 0.000 description 7
- 239000000523 sample Substances 0.000 description 7
- 239000011780 sodium chloride Substances 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 6
- 238000002835 absorbance Methods 0.000 description 6
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 6
- 230000000890 antigenic effect Effects 0.000 description 6
- 239000013604 expression vector Substances 0.000 description 6
- 230000006870 function Effects 0.000 description 6
- 238000004128 high performance liquid chromatography Methods 0.000 description 6
- 108091008146 restriction endonucleases Proteins 0.000 description 6
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 230000009870 specific binding Effects 0.000 description 6
- 239000006228 supernatant Substances 0.000 description 6
- 108010003137 tyrosyltyrosine Proteins 0.000 description 6
- 102000014914 Carrier Proteins Human genes 0.000 description 5
- 108091028043 Nucleic acid sequence Proteins 0.000 description 5
- IHCXPSYCHXFXKT-DCAQKATOSA-N Pro-Arg-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O IHCXPSYCHXFXKT-DCAQKATOSA-N 0.000 description 5
- YMTLKLXDFCSCNX-BYPYZUCNSA-N Ser-Gly-Gly Chemical compound OC[C@H](N)C(=O)NCC(=O)NCC(O)=O YMTLKLXDFCSCNX-BYPYZUCNSA-N 0.000 description 5
- 238000001042 affinity chromatography Methods 0.000 description 5
- 108010087924 alanylproline Proteins 0.000 description 5
- 108091008324 binding proteins Proteins 0.000 description 5
- 239000003795 chemical substances by application Substances 0.000 description 5
- 210000004978 chinese hamster ovary cell Anatomy 0.000 description 5
- 238000010586 diagram Methods 0.000 description 5
- 238000010494 dissociation reaction Methods 0.000 description 5
- 230000005593 dissociations Effects 0.000 description 5
- 239000000499 gel Substances 0.000 description 5
- BRZYSWJRSDMWLG-CAXSIQPQSA-N geneticin Chemical compound O1C[C@@](O)(C)[C@H](NC)[C@@H](O)[C@H]1O[C@@H]1[C@@H](O)[C@H](O[C@@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](C(C)O)O2)N)[C@@H](N)C[C@H]1N BRZYSWJRSDMWLG-CAXSIQPQSA-N 0.000 description 5
- 210000004602 germ cell Anatomy 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000000746 purification Methods 0.000 description 5
- 238000001542 size-exclusion chromatography Methods 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 4
- 229920000936 Agarose Polymers 0.000 description 4
- SHZGCJCMOBCMKK-UHFFFAOYSA-N D-mannomethylose Natural products CC1OC(O)C(O)C(O)C1O SHZGCJCMOBCMKK-UHFFFAOYSA-N 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 4
- 241000588724 Escherichia coli Species 0.000 description 4
- PNNNRSAQSRJVSB-SLPGGIOYSA-N Fucose Natural products C[C@H](O)[C@@H](O)[C@H](O)[C@H](O)C=O PNNNRSAQSRJVSB-SLPGGIOYSA-N 0.000 description 4
- UGTHTQWIQKEDEH-BQBZGAKWSA-N L-alanyl-L-prolylglycine zwitterion Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O UGTHTQWIQKEDEH-BQBZGAKWSA-N 0.000 description 4
- SHZGCJCMOBCMKK-DHVFOXMCSA-N L-fucopyranose Chemical compound C[C@@H]1OC(O)[C@@H](O)[C@H](O)[C@@H]1O SHZGCJCMOBCMKK-DHVFOXMCSA-N 0.000 description 4
- TYYLDKGBCJGJGW-UHFFFAOYSA-N L-tryptophan-L-tyrosine Natural products C=1NC2=CC=CC=C2C=1CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 TYYLDKGBCJGJGW-UHFFFAOYSA-N 0.000 description 4
- AIMGJYMCTAABEN-GVXVVHGQSA-N Leu-Val-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O AIMGJYMCTAABEN-GVXVVHGQSA-N 0.000 description 4
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 4
- ZJPGOXWRFNKIQL-JYJNAYRXSA-N Phe-Pro-Pro Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(O)=O)C1=CC=CC=C1 ZJPGOXWRFNKIQL-JYJNAYRXSA-N 0.000 description 4
- 108020004511 Recombinant DNA Proteins 0.000 description 4
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-N 0.000 description 4
- SRSPTFBENMJHMR-WHFBIAKZSA-N Ser-Ser-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O SRSPTFBENMJHMR-WHFBIAKZSA-N 0.000 description 4
- BMKNXTJLHFIAAH-CIUDSAMLSA-N Ser-Ser-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O BMKNXTJLHFIAAH-CIUDSAMLSA-N 0.000 description 4
- XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 description 4
- HTONZBWRYUKUKC-RCWTZXSCSA-N Val-Thr-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O HTONZBWRYUKUKC-RCWTZXSCSA-N 0.000 description 4
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 4
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 4
- 108010050025 alpha-glutamyltryptophan Proteins 0.000 description 4
- 108010092854 aspartyllysine Proteins 0.000 description 4
- 238000003556 assay Methods 0.000 description 4
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 4
- 239000011324 bead Substances 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 4
- 239000003623 enhancer Substances 0.000 description 4
- 210000003527 eukaryotic cell Anatomy 0.000 description 4
- 108010089804 glycyl-threonine Proteins 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 230000001900 immune effect Effects 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 239000003446 ligand Substances 0.000 description 4
- 230000001404 mediated effect Effects 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 230000005180 public health Effects 0.000 description 4
- 238000010188 recombinant method Methods 0.000 description 4
- 238000013207 serial dilution Methods 0.000 description 4
- 238000010561 standard procedure Methods 0.000 description 4
- 108010044292 tryptophyltyrosine Proteins 0.000 description 4
- NFGXHKASABOEEW-UHFFFAOYSA-N 1-methylethyl 11-methoxy-3,7,11-trimethyl-2,4-dodecadienoate Chemical compound COC(C)(C)CCCC(C)CC=CC(C)=CC(=O)OC(C)C NFGXHKASABOEEW-UHFFFAOYSA-N 0.000 description 3
- XWFWAXPOLRTDFZ-FXQIFTODSA-N Ala-Pro-Ser Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O XWFWAXPOLRTDFZ-FXQIFTODSA-N 0.000 description 3
- HPBNLFLSSQDFQW-WHFBIAKZSA-N Asn-Ser-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O HPBNLFLSSQDFQW-WHFBIAKZSA-N 0.000 description 3
- YNQIDCRRTWGHJD-ZLUOBGJFSA-N Asp-Asn-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC(O)=O YNQIDCRRTWGHJD-ZLUOBGJFSA-N 0.000 description 3
- VNXQRBXEQXLERQ-CIUDSAMLSA-N Asp-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)O)N VNXQRBXEQXLERQ-CIUDSAMLSA-N 0.000 description 3
- YIDFBWRHIYOYAA-LKXGYXEUSA-N Asp-Ser-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YIDFBWRHIYOYAA-LKXGYXEUSA-N 0.000 description 3
- MNQMTYSEKZHIDF-GCJQMDKQSA-N Asp-Thr-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O MNQMTYSEKZHIDF-GCJQMDKQSA-N 0.000 description 3
- UHOVQNZJYSORNB-UHFFFAOYSA-N Benzene Chemical compound C1=CC=CC=C1 UHOVQNZJYSORNB-UHFFFAOYSA-N 0.000 description 3
- 238000001712 DNA sequencing Methods 0.000 description 3
- DMYACXMQUABZIQ-NRPADANISA-N Glu-Ser-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O DMYACXMQUABZIQ-NRPADANISA-N 0.000 description 3
- ZYRXTRTUCAVNBQ-GVXVVHGQSA-N Glu-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N ZYRXTRTUCAVNBQ-GVXVVHGQSA-N 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- NNCSJUBVFBDDLC-YUMQZZPRSA-N Gly-Leu-Ser Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O NNCSJUBVFBDDLC-YUMQZZPRSA-N 0.000 description 3
- GBYYQVBXFVDJPJ-WLTAIBSBSA-N Gly-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)CN)O GBYYQVBXFVDJPJ-WLTAIBSBSA-N 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 239000004471 Glycine Substances 0.000 description 3
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 3
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 3
- 108700005091 Immunoglobulin Genes Proteins 0.000 description 3
- PVMPDMIKUVNOBD-CIUDSAMLSA-N Leu-Asp-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O PVMPDMIKUVNOBD-CIUDSAMLSA-N 0.000 description 3
- VCHVSKNMTXWIIP-SRVKXCTJSA-N Leu-Lys-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O VCHVSKNMTXWIIP-SRVKXCTJSA-N 0.000 description 3
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 3
- PDIDTSZKKFEDMB-UWVGGRQHSA-N Lys-Pro-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O PDIDTSZKKFEDMB-UWVGGRQHSA-N 0.000 description 3
- AJHCSUXXECOXOY-UHFFFAOYSA-N N-glycyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-UHFFFAOYSA-N 0.000 description 3
- 239000004793 Polystyrene Substances 0.000 description 3
- PRKWBYCXBBSLSK-GUBZILKMSA-N Pro-Ser-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O PRKWBYCXBBSLSK-GUBZILKMSA-N 0.000 description 3
- 229920002684 Sepharose Polymers 0.000 description 3
- FTVRVZNYIYWJGB-ACZMJKKPSA-N Ser-Asp-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O FTVRVZNYIYWJGB-ACZMJKKPSA-N 0.000 description 3
- BPMRXBZYPGYPJN-WHFBIAKZSA-N Ser-Gly-Asn Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O BPMRXBZYPGYPJN-WHFBIAKZSA-N 0.000 description 3
- GZSZPKSBVAOGIE-CIUDSAMLSA-N Ser-Lys-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O GZSZPKSBVAOGIE-CIUDSAMLSA-N 0.000 description 3
- CKDXFSPMIDSMGV-GUBZILKMSA-N Ser-Pro-Val Chemical compound [H]N[C@@H](CO)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O CKDXFSPMIDSMGV-GUBZILKMSA-N 0.000 description 3
- DXPURPNJDFCKKO-RHYQMDGZSA-N Thr-Lys-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)[C@@H](C)O)C(O)=O DXPURPNJDFCKKO-RHYQMDGZSA-N 0.000 description 3
- UYKREHOKELZSPB-JTQLQIEISA-N Trp-Gly Chemical compound C1=CC=C2C(C[C@H](N)C(=O)NCC(O)=O)=CNC2=C1 UYKREHOKELZSPB-JTQLQIEISA-N 0.000 description 3
- NLWCSMOXNKBRLC-WDSOQIARSA-N Trp-Lys-Val Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O NLWCSMOXNKBRLC-WDSOQIARSA-N 0.000 description 3
- MBLJBGZWLHTJBH-SZMVWBNQSA-N Trp-Val-Arg Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)=CNC2=C1 MBLJBGZWLHTJBH-SZMVWBNQSA-N 0.000 description 3
- MQGGXGKQSVEQHR-KKUMJFAQSA-N Tyr-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 MQGGXGKQSVEQHR-KKUMJFAQSA-N 0.000 description 3
- SSYBNWFXCFNRFN-GUBZILKMSA-N Val-Pro-Ser Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SSYBNWFXCFNRFN-GUBZILKMSA-N 0.000 description 3
- OWFGFHQMSBTKLX-UFYCRDLUSA-N Val-Tyr-Tyr Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)O)N OWFGFHQMSBTKLX-UFYCRDLUSA-N 0.000 description 3
- 108010073929 Vascular Endothelial Growth Factor A Proteins 0.000 description 3
- 230000004071 biological effect Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 230000003013 cytotoxicity Effects 0.000 description 3
- 231100000135 cytotoxicity Toxicity 0.000 description 3
- 230000001419 dependent effect Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000001502 gel electrophoresis Methods 0.000 description 3
- 238000007429 general method Methods 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 150000004676 glycans Chemical class 0.000 description 3
- 230000013595 glycosylation Effects 0.000 description 3
- 238000006206 glycosylation reaction Methods 0.000 description 3
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 3
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 3
- 108010050475 glycyl-leucyl-tyrosine Proteins 0.000 description 3
- 108010084389 glycyltryptophan Proteins 0.000 description 3
- 229940072221 immunoglobulins Drugs 0.000 description 3
- 230000001965 increasing effect Effects 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000003993 interaction Effects 0.000 description 3
- 201000005202 lung cancer Diseases 0.000 description 3
- 208000020816 lung neoplasm Diseases 0.000 description 3
- 108010017391 lysylvaline Proteins 0.000 description 3
- 238000004949 mass spectrometry Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 3
- 239000002773 nucleotide Substances 0.000 description 3
- 125000003729 nucleotide group Chemical group 0.000 description 3
- 108010024654 phenylalanyl-prolyl-alanine Proteins 0.000 description 3
- 108010051242 phenylalanylserine Proteins 0.000 description 3
- 230000026731 phosphorylation Effects 0.000 description 3
- 238000006366 phosphorylation reaction Methods 0.000 description 3
- 229920002223 polystyrene Polymers 0.000 description 3
- 210000001236 prokaryotic cell Anatomy 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 108010031719 prolyl-serine Proteins 0.000 description 3
- 239000011347 resin Substances 0.000 description 3
- 229920005989 resin Polymers 0.000 description 3
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 238000001890 transfection Methods 0.000 description 3
- 230000010474 transient expression Effects 0.000 description 3
- 108010038745 tryptophylglycine Proteins 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 108010071635 tyrosyl-prolyl-arginine Proteins 0.000 description 3
- HZAXFHJVJLSVMW-UHFFFAOYSA-N 2-Aminoethan-1-ol Chemical compound NCCO HZAXFHJVJLSVMW-UHFFFAOYSA-N 0.000 description 2
- DKJPOZOEBONHFS-ZLUOBGJFSA-N Ala-Ala-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O DKJPOZOEBONHFS-ZLUOBGJFSA-N 0.000 description 2
- YYSWCHMLFJLLBJ-ZLUOBGJFSA-N Ala-Ala-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YYSWCHMLFJLLBJ-ZLUOBGJFSA-N 0.000 description 2
- MNZHHDPWDWQJCQ-YUMQZZPRSA-N Ala-Leu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O MNZHHDPWDWQJCQ-YUMQZZPRSA-N 0.000 description 2
- MEFILNJXAVSUTO-JXUBOQSCSA-N Ala-Leu-Thr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MEFILNJXAVSUTO-JXUBOQSCSA-N 0.000 description 2
- MDNAVFBZPROEHO-DCAQKATOSA-N Ala-Lys-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O MDNAVFBZPROEHO-DCAQKATOSA-N 0.000 description 2
- MDNAVFBZPROEHO-UHFFFAOYSA-N Ala-Lys-Val Natural products CC(C)C(C(O)=O)NC(=O)C(NC(=O)C(C)N)CCCCN MDNAVFBZPROEHO-UHFFFAOYSA-N 0.000 description 2
- WQKAQKZRDIZYNV-VZFHVOOUSA-N Ala-Ser-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WQKAQKZRDIZYNV-VZFHVOOUSA-N 0.000 description 2
- YJHKTAMKPGFJCT-NRPADANISA-N Ala-Val-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O YJHKTAMKPGFJCT-NRPADANISA-N 0.000 description 2
- 101710145634 Antigen 1 Proteins 0.000 description 2
- VKKYFICVTYKFIO-CIUDSAMLSA-N Arg-Ala-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N VKKYFICVTYKFIO-CIUDSAMLSA-N 0.000 description 2
- YSUVMPICYVWRBX-VEVYYDQMSA-N Arg-Asp-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O YSUVMPICYVWRBX-VEVYYDQMSA-N 0.000 description 2
- OLVIPTLKNSAYRJ-YUMQZZPRSA-N Asn-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N OLVIPTLKNSAYRJ-YUMQZZPRSA-N 0.000 description 2
- QUAWOKPCAKCHQL-SRVKXCTJSA-N Asn-His-Lys Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N QUAWOKPCAKCHQL-SRVKXCTJSA-N 0.000 description 2
- RBOBTTLFPRSXKZ-BZSNNMDCSA-N Asn-Phe-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O RBOBTTLFPRSXKZ-BZSNNMDCSA-N 0.000 description 2
- JTXVXGXTRXMOFJ-FXQIFTODSA-N Asn-Pro-Asn Chemical compound NC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O JTXVXGXTRXMOFJ-FXQIFTODSA-N 0.000 description 2
- KZYSHAMXEBPJBD-JRQIVUDYSA-N Asn-Thr-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KZYSHAMXEBPJBD-JRQIVUDYSA-N 0.000 description 2
- SNDBKTFJWVEVPO-WHFBIAKZSA-N Asp-Gly-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](CO)C(O)=O SNDBKTFJWVEVPO-WHFBIAKZSA-N 0.000 description 2
- YUELDQUPTAYEGM-XIRDDKMYSA-N Asp-Trp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CC(=O)O)N YUELDQUPTAYEGM-XIRDDKMYSA-N 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 102000012422 Collagen Type I Human genes 0.000 description 2
- 108010022452 Collagen Type I Proteins 0.000 description 2
- 108010047041 Complementarity Determining Regions Proteins 0.000 description 2
- NDUSUIGBMZCOIL-ZKWXMUAHSA-N Cys-Asn-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CS)N NDUSUIGBMZCOIL-ZKWXMUAHSA-N 0.000 description 2
- OHLLDUNVMPPUMD-DCAQKATOSA-N Cys-Leu-Val Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CS)N OHLLDUNVMPPUMD-DCAQKATOSA-N 0.000 description 2
- ZXCAQANTQWBICD-DCAQKATOSA-N Cys-Lys-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CS)N ZXCAQANTQWBICD-DCAQKATOSA-N 0.000 description 2
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical group OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 2
- 238000012286 ELISA Assay Methods 0.000 description 2
- 102000009109 Fc receptors Human genes 0.000 description 2
- 108010087819 Fc receptors Proteins 0.000 description 2
- QDMVXRNLOPTPIE-WDCWCFNPSA-N Glu-Lys-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QDMVXRNLOPTPIE-WDCWCFNPSA-N 0.000 description 2
- MIWJDJAMMKHUAR-ZVZYQTTQSA-N Glu-Trp-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CCC(=O)O)N MIWJDJAMMKHUAR-ZVZYQTTQSA-N 0.000 description 2
- MFYLRRCYBBJYPI-JYJNAYRXSA-N Glu-Tyr-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O MFYLRRCYBBJYPI-JYJNAYRXSA-N 0.000 description 2
- WKJKBELXHCTHIJ-WPRPVWTQSA-N Gly-Arg-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N WKJKBELXHCTHIJ-WPRPVWTQSA-N 0.000 description 2
- BIRKKBCSAIHDDF-WDSKDSINSA-N Gly-Glu-Cys Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CS)C(O)=O BIRKKBCSAIHDDF-WDSKDSINSA-N 0.000 description 2
- UQJNXZSSGQIPIQ-FBCQKBJTSA-N Gly-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)CN UQJNXZSSGQIPIQ-FBCQKBJTSA-N 0.000 description 2
- ULZCYBYDTUMHNF-IUCAKERBSA-N Gly-Leu-Glu Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O ULZCYBYDTUMHNF-IUCAKERBSA-N 0.000 description 2
- MIIVFRCYJABHTQ-ONGXEEELSA-N Gly-Leu-Val Chemical compound [H]NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O MIIVFRCYJABHTQ-ONGXEEELSA-N 0.000 description 2
- FFJQHWKSGAWSTJ-BFHQHQDPSA-N Gly-Thr-Ala Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O FFJQHWKSGAWSTJ-BFHQHQDPSA-N 0.000 description 2
- ZZWUYQXMIFTIIY-WEDXCCLWSA-N Gly-Thr-Leu Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O ZZWUYQXMIFTIIY-WEDXCCLWSA-N 0.000 description 2
- BXDLTKLPPKBVEL-FJXKBIBVSA-N Gly-Thr-Met Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCSC)C(O)=O BXDLTKLPPKBVEL-FJXKBIBVSA-N 0.000 description 2
- AJHCSUXXECOXOY-NSHDSACASA-N Gly-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)CN)C(O)=O)=CNC2=C1 AJHCSUXXECOXOY-NSHDSACASA-N 0.000 description 2
- 102000051366 Glycosyltransferases Human genes 0.000 description 2
- 108700023372 Glycosyltransferases Proteins 0.000 description 2
- ZRALSGWEFCBTJO-UHFFFAOYSA-N Guanidine Chemical compound NC(N)=N ZRALSGWEFCBTJO-UHFFFAOYSA-N 0.000 description 2
- TVMNTHXFRSXZGR-IHRRRGAJSA-N His-Lys-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O TVMNTHXFRSXZGR-IHRRRGAJSA-N 0.000 description 2
- 101000808011 Homo sapiens Vascular endothelial growth factor A Proteins 0.000 description 2
- 108010021625 Immunoglobulin Fragments Proteins 0.000 description 2
- 102000008394 Immunoglobulin Fragments Human genes 0.000 description 2
- 108010065920 Insulin Lispro Proteins 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- SENJXOPIZNYLHU-UHFFFAOYSA-N L-leucyl-L-arginine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CCCN=C(N)N SENJXOPIZNYLHU-UHFFFAOYSA-N 0.000 description 2
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 2
- IAJFFZORSWOZPQ-SRVKXCTJSA-N Leu-Leu-Asn Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IAJFFZORSWOZPQ-SRVKXCTJSA-N 0.000 description 2
- DPURXCQCHSQPAN-AVGNSLFASA-N Leu-Pro-Pro Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DPURXCQCHSQPAN-AVGNSLFASA-N 0.000 description 2
- KIZIOFNVSOSKJI-CIUDSAMLSA-N Leu-Ser-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N KIZIOFNVSOSKJI-CIUDSAMLSA-N 0.000 description 2
- SBANPBVRHYIMRR-UHFFFAOYSA-N Leu-Ser-Pro Natural products CC(C)CC(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O SBANPBVRHYIMRR-UHFFFAOYSA-N 0.000 description 2
- BRTVHXHCUSXYRI-CIUDSAMLSA-N Leu-Ser-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O BRTVHXHCUSXYRI-CIUDSAMLSA-N 0.000 description 2
- VUBIPAHVHMZHCM-KKUMJFAQSA-N Leu-Tyr-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CO)C(O)=O)CC1=CC=C(O)C=C1 VUBIPAHVHMZHCM-KKUMJFAQSA-N 0.000 description 2
- MPGHETGWWWUHPY-CIUDSAMLSA-N Lys-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN MPGHETGWWWUHPY-CIUDSAMLSA-N 0.000 description 2
- NTBFKPBULZGXQL-KKUMJFAQSA-N Lys-Asp-Tyr Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 NTBFKPBULZGXQL-KKUMJFAQSA-N 0.000 description 2
- GQFDWEDHOQRNLC-QWRGUYRKSA-N Lys-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN GQFDWEDHOQRNLC-QWRGUYRKSA-N 0.000 description 2
- RFQATBGBLDAKGI-VHSXEESVSA-N Lys-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CCCCN)N)C(=O)O RFQATBGBLDAKGI-VHSXEESVSA-N 0.000 description 2
- QQPSCXKFDSORFT-IHRRRGAJSA-N Lys-Lys-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CCCCN QQPSCXKFDSORFT-IHRRRGAJSA-N 0.000 description 2
- QCZYYEFXOBKCNQ-STQMWFEESA-N Lys-Phe Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 QCZYYEFXOBKCNQ-STQMWFEESA-N 0.000 description 2
- IOQWIOPSKJOEKI-SRVKXCTJSA-N Lys-Ser-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O IOQWIOPSKJOEKI-SRVKXCTJSA-N 0.000 description 2
- MIFFFXHMAHFACR-KATARQTJSA-N Lys-Ser-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CCCCN MIFFFXHMAHFACR-KATARQTJSA-N 0.000 description 2
- 239000007987 MES buffer Substances 0.000 description 2
- 239000007993 MOPS buffer Substances 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241001529936 Murinae Species 0.000 description 2
- 241000699666 Mus <mouse, genus> Species 0.000 description 2
- LRHPLDYGYMQRHN-UHFFFAOYSA-N N-Butanol Chemical compound CCCCO LRHPLDYGYMQRHN-UHFFFAOYSA-N 0.000 description 2
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 2
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 2
- 108010079364 N-glycylalanine Proteins 0.000 description 2
- 108091034117 Oligonucleotide Proteins 0.000 description 2
- 239000012124 Opti-MEM Substances 0.000 description 2
- HXSUFWQYLPKEHF-IHRRRGAJSA-N Phe-Asn-Arg Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N HXSUFWQYLPKEHF-IHRRRGAJSA-N 0.000 description 2
- QARPMYDMYVLFMW-KKUMJFAQSA-N Phe-Pro-Glu Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(O)=O)C1=CC=CC=C1 QARPMYDMYVLFMW-KKUMJFAQSA-N 0.000 description 2
- JHSRGEODDALISP-XVSYOHENSA-N Phe-Thr-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O JHSRGEODDALISP-XVSYOHENSA-N 0.000 description 2
- BPIMVBKDLSBKIJ-FCLVOEFKSA-N Phe-Thr-Phe Chemical compound C([C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 BPIMVBKDLSBKIJ-FCLVOEFKSA-N 0.000 description 2
- 102000029797 Prion Human genes 0.000 description 2
- 108091000054 Prion Proteins 0.000 description 2
- VPEVBAUSTBWQHN-NHCYSSNCSA-N Pro-Glu-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O VPEVBAUSTBWQHN-NHCYSSNCSA-N 0.000 description 2
- HAAQQNHQZBOWFO-LURJTMIESA-N Pro-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H]1CCCN1 HAAQQNHQZBOWFO-LURJTMIESA-N 0.000 description 2
- UIMCLYYSUCIUJM-UWVGGRQHSA-N Pro-Gly-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H]1CCCN1 UIMCLYYSUCIUJM-UWVGGRQHSA-N 0.000 description 2
- XFFIGWGYMUFCCQ-ULQDDVLXSA-N Pro-His-Tyr Chemical compound C1=CC(O)=CC=C1C[C@@H](C([O-])=O)NC(=O)[C@@H](NC(=O)[C@H]1[NH2+]CCC1)CC1=CN=CN1 XFFIGWGYMUFCCQ-ULQDDVLXSA-N 0.000 description 2
- FMLRRBDLBJLJIK-DCAQKATOSA-N Pro-Leu-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]1CCCN1 FMLRRBDLBJLJIK-DCAQKATOSA-N 0.000 description 2
- MHHQQZIFLWFZGR-DCAQKATOSA-N Pro-Lys-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O MHHQQZIFLWFZGR-DCAQKATOSA-N 0.000 description 2
- OWQXAJQZLWHPBH-FXQIFTODSA-N Pro-Ser-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O OWQXAJQZLWHPBH-FXQIFTODSA-N 0.000 description 2
- MKGIILKDUGDRRO-FXQIFTODSA-N Pro-Ser-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H]1CCCN1 MKGIILKDUGDRRO-FXQIFTODSA-N 0.000 description 2
- VVAWNPIOYXAMAL-KJEVXHAQSA-N Pro-Thr-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VVAWNPIOYXAMAL-KJEVXHAQSA-N 0.000 description 2
- YDTUEBLEAVANFH-RCWTZXSCSA-N Pro-Val-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 YDTUEBLEAVANFH-RCWTZXSCSA-N 0.000 description 2
- 206010036790 Productive cough Diseases 0.000 description 2
- 108010076504 Protein Sorting Signals Proteins 0.000 description 2
- OYEDZGNMSBZCIM-XGEHTFHBSA-N Ser-Arg-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OYEDZGNMSBZCIM-XGEHTFHBSA-N 0.000 description 2
- UQFYNFTYDHUIMI-WHFBIAKZSA-N Ser-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H](N)CO UQFYNFTYDHUIMI-WHFBIAKZSA-N 0.000 description 2
- XXXAXOWMBOKTRN-XPUUQOCRSA-N Ser-Gly-Val Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O XXXAXOWMBOKTRN-XPUUQOCRSA-N 0.000 description 2
- NBUKGEFVZJMSIS-XIRDDKMYSA-N Ser-His-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC3=CN=CN3)NC(=O)[C@H](CO)N NBUKGEFVZJMSIS-XIRDDKMYSA-N 0.000 description 2
- QYSFWUIXDFJUDW-DCAQKATOSA-N Ser-Leu-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QYSFWUIXDFJUDW-DCAQKATOSA-N 0.000 description 2
- NLOAIFSWUUFQFR-CIUDSAMLSA-N Ser-Leu-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O NLOAIFSWUUFQFR-CIUDSAMLSA-N 0.000 description 2
- HHJFMHQYEAAOBM-ZLUOBGJFSA-N Ser-Ser-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O HHJFMHQYEAAOBM-ZLUOBGJFSA-N 0.000 description 2
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 2
- PYTKULIABVRXSC-BWBBJGPYSA-N Ser-Ser-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PYTKULIABVRXSC-BWBBJGPYSA-N 0.000 description 2
- PMTWIUBUQRGCSB-FXQIFTODSA-N Ser-Val-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](C)C(O)=O PMTWIUBUQRGCSB-FXQIFTODSA-N 0.000 description 2
- HNDMFDBQXYZSRM-IHRRRGAJSA-N Ser-Val-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HNDMFDBQXYZSRM-IHRRRGAJSA-N 0.000 description 2
- HSWXBJCBYSWBPT-GUBZILKMSA-N Ser-Val-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CO)C(C)C)C(O)=O HSWXBJCBYSWBPT-GUBZILKMSA-N 0.000 description 2
- 239000004268 Sodium erythorbin Substances 0.000 description 2
- 208000005718 Stomach Neoplasms Diseases 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 2
- 239000012505 Superdex™ Substances 0.000 description 2
- JVTHIXKSVYEWNI-JRQIVUDYSA-N Thr-Asn-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JVTHIXKSVYEWNI-JRQIVUDYSA-N 0.000 description 2
- AQAMPXBRJJWPNI-JHEQGTHGSA-N Thr-Gly-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AQAMPXBRJJWPNI-JHEQGTHGSA-N 0.000 description 2
- YOOAQCZYZHGUAZ-KATARQTJSA-N Thr-Leu-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YOOAQCZYZHGUAZ-KATARQTJSA-N 0.000 description 2
- XSEPSRUDSPHMPX-KATARQTJSA-N Thr-Lys-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O XSEPSRUDSPHMPX-KATARQTJSA-N 0.000 description 2
- KPNSNVTUVKSBFL-ZJDVBMNYSA-N Thr-Met-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KPNSNVTUVKSBFL-ZJDVBMNYSA-N 0.000 description 2
- JMBRNXUOLJFURW-BEAPCOKYSA-N Thr-Phe-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N)O JMBRNXUOLJFURW-BEAPCOKYSA-N 0.000 description 2
- IQPWNQRRAJHOKV-KATARQTJSA-N Thr-Ser-Lys Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN IQPWNQRRAJHOKV-KATARQTJSA-N 0.000 description 2
- WCRFXRIWBFRZBR-GGVZMXCHSA-N Thr-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 WCRFXRIWBFRZBR-GGVZMXCHSA-N 0.000 description 2
- BEZTUFWTPVOROW-KJEVXHAQSA-N Thr-Tyr-Arg Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N)O BEZTUFWTPVOROW-KJEVXHAQSA-N 0.000 description 2
- 239000007983 Tris buffer Substances 0.000 description 2
- UDCHKDYNMRJYMI-QEJZJMRPSA-N Trp-Glu-Ser Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O UDCHKDYNMRJYMI-QEJZJMRPSA-N 0.000 description 2
- QJBWZNTWJSZUOY-UWJYBYFXSA-N Tyr-Ala-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N QJBWZNTWJSZUOY-UWJYBYFXSA-N 0.000 description 2
- SCCKSNREWHMKOJ-SRVKXCTJSA-N Tyr-Asn-Ser Chemical compound N[C@@H](Cc1ccc(O)cc1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O SCCKSNREWHMKOJ-SRVKXCTJSA-N 0.000 description 2
- SLCSPPCQWUHPPO-JYJNAYRXSA-N Tyr-Glu-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 SLCSPPCQWUHPPO-JYJNAYRXSA-N 0.000 description 2
- AZGZDDNKFFUDEH-QWRGUYRKSA-N Tyr-Gly-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 AZGZDDNKFFUDEH-QWRGUYRKSA-N 0.000 description 2
- LMKKMCGTDANZTR-BZSNNMDCSA-N Tyr-Phe-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 LMKKMCGTDANZTR-BZSNNMDCSA-N 0.000 description 2
- RMRFSFXLFWWAJZ-HJOGWXRNSA-N Tyr-Tyr-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 RMRFSFXLFWWAJZ-HJOGWXRNSA-N 0.000 description 2
- SYSWVVCYSXBVJG-RHYQMDGZSA-N Val-Leu-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](C(C)C)N)O SYSWVVCYSXBVJG-RHYQMDGZSA-N 0.000 description 2
- VHIZXDZMTDVFGX-DCAQKATOSA-N Val-Ser-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N VHIZXDZMTDVFGX-DCAQKATOSA-N 0.000 description 2
- SDHZOOIGIUEPDY-JYJNAYRXSA-N Val-Ser-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CO)NC(=O)[C@@H](N)C(C)C)C(O)=O)=CNC2=C1 SDHZOOIGIUEPDY-JYJNAYRXSA-N 0.000 description 2
- TVGWMCTYUFBXAP-QTKMDUPCSA-N Val-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C(C)C)N)O TVGWMCTYUFBXAP-QTKMDUPCSA-N 0.000 description 2
- ZHWZDZFWBXWPDW-GUBZILKMSA-N Val-Val-Cys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CS)C(O)=O ZHWZDZFWBXWPDW-GUBZILKMSA-N 0.000 description 2
- LLJLBRRXKZTTRD-GUBZILKMSA-N Val-Val-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)O)N LLJLBRRXKZTTRD-GUBZILKMSA-N 0.000 description 2
- JVGDAEKKZKKZFO-RCWTZXSCSA-N Val-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](C(C)C)N)O JVGDAEKKZKKZFO-RCWTZXSCSA-N 0.000 description 2
- 241000700605 Viruses Species 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000000996 additive effect Effects 0.000 description 2
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 2
- 108010041407 alanylaspartic acid Proteins 0.000 description 2
- 108010070783 alanyltyrosine Proteins 0.000 description 2
- 239000012491 analyte Substances 0.000 description 2
- 239000003429 antifungal agent Substances 0.000 description 2
- 229940121375 antifungal agent Drugs 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 108010009111 arginyl-glycyl-glutamic acid Proteins 0.000 description 2
- OWMVSZAMULFTJU-UHFFFAOYSA-N bis-tris Chemical compound OCCN(CCO)C(CO)(CO)CO OWMVSZAMULFTJU-UHFFFAOYSA-N 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- AIYUHDOJVYHVIT-UHFFFAOYSA-M caesium chloride Chemical compound [Cl-].[Cs+] AIYUHDOJVYHVIT-UHFFFAOYSA-M 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 235000014633 carbohydrates Nutrition 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 238000012832 cell culture technique Methods 0.000 description 2
- 210000002421 cell wall Anatomy 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 210000003169 central nervous system Anatomy 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 230000024203 complement activation Effects 0.000 description 2
- 108010060199 cysteinylproline Proteins 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 230000005714 functional activity Effects 0.000 description 2
- 206010017758 gastric cancer Diseases 0.000 description 2
- 108010013768 glutamyl-aspartyl-proline Proteins 0.000 description 2
- 108010049041 glutamylalanine Proteins 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 2
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 238000005734 heterodimerization reaction Methods 0.000 description 2
- 102000058223 human VEGFA Human genes 0.000 description 2
- 210000004408 hybridoma Anatomy 0.000 description 2
- 230000003053 immunization Effects 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000001802 infusion Methods 0.000 description 2
- 229940065638 intron a Drugs 0.000 description 2
- 239000007951 isotonicity adjuster Substances 0.000 description 2
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 2
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 2
- 108010000761 leucylarginine Proteins 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 108010038320 lysylphenylalanine Proteins 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 108020004999 messenger RNA Proteins 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000000178 monomer Substances 0.000 description 2
- 238000002823 phage display Methods 0.000 description 2
- YBYRMVIVWMBXKQ-UHFFFAOYSA-N phenylmethanesulfonyl fluoride Chemical compound FS(=O)(=O)CC1=CC=CC=C1 YBYRMVIVWMBXKQ-UHFFFAOYSA-N 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 230000008488 polyadenylation Effects 0.000 description 2
- 108091033319 polynucleotide Proteins 0.000 description 2
- 102000040430 polynucleotide Human genes 0.000 description 2
- 239000002157 polynucleotide Substances 0.000 description 2
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 2
- 108010070643 prolylglutamic acid Proteins 0.000 description 2
- 238000001742 protein purification Methods 0.000 description 2
- 230000004850 protein–protein interaction Effects 0.000 description 2
- 102000005962 receptors Human genes 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 238000003259 recombinant expression Methods 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 2
- 230000000392 somatic effect Effects 0.000 description 2
- 210000003802 sputum Anatomy 0.000 description 2
- 208000024794 sputum Diseases 0.000 description 2
- 201000011549 stomach cancer Diseases 0.000 description 2
- 238000007920 subcutaneous administration Methods 0.000 description 2
- 229910021653 sulphate ion Inorganic materials 0.000 description 2
- 238000002198 surface plasmon resonance spectroscopy Methods 0.000 description 2
- 108010072986 threonyl-seryl-lysine Proteins 0.000 description 2
- 238000013518 transcription Methods 0.000 description 2
- 230000035897 transcription Effects 0.000 description 2
- 239000012096 transfection reagent Substances 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 238000003146 transient transfection Methods 0.000 description 2
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 2
- 108010079202 tyrosyl-alanyl-cysteine Proteins 0.000 description 2
- 108010073969 valyllysine Proteins 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- IESDGNYHXIOKRW-YXMSTPNBSA-N (2s)-2-[[(2s)-1-[(2s)-6-amino-2-[[(2s,3r)-2-amino-3-hydroxybutanoyl]amino]hexanoyl]pyrrolidine-2-carbonyl]amino]-5-(diaminomethylideneamino)pentanoic acid Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O IESDGNYHXIOKRW-YXMSTPNBSA-N 0.000 description 1
- DIBLBAURNYJYBF-XLXZRNDBSA-N (2s)-2-[[(2s)-2-[[2-[[(2s)-6-amino-2-[[(2s)-2-amino-3-methylbutanoyl]amino]hexanoyl]amino]acetyl]amino]-3-phenylpropanoyl]amino]-3-(4-hydroxyphenyl)propanoic acid Chemical compound C([C@H](NC(=O)CNC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)C(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 DIBLBAURNYJYBF-XLXZRNDBSA-N 0.000 description 1
- HNSDLXPSAYFUHK-UHFFFAOYSA-N 1,4-bis(2-ethylhexyl) sulfosuccinate Chemical compound CCCCC(CC)COC(=O)CC(S(O)(=O)=O)C(=O)OCC(CC)CCCC HNSDLXPSAYFUHK-UHFFFAOYSA-N 0.000 description 1
- AEQDJSLRWYMAQI-UHFFFAOYSA-N 2,3,9,10-tetramethoxy-6,8,13,13a-tetrahydro-5H-isoquinolino[2,1-b]isoquinoline Chemical compound C1CN2CC(C(=C(OC)C=C3)OC)=C3CC2C2=C1C=C(OC)C(OC)=C2 AEQDJSLRWYMAQI-UHFFFAOYSA-N 0.000 description 1
- JKMHFZQWWAIEOD-UHFFFAOYSA-N 2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid Chemical compound OCC[NH+]1CCN(CCS([O-])(=O)=O)CC1 JKMHFZQWWAIEOD-UHFFFAOYSA-N 0.000 description 1
- 125000000022 2-aminoethyl group Chemical group [H]C([*])([H])C([H])([H])N([H])[H] 0.000 description 1
- QFVHZQCOUORWEI-UHFFFAOYSA-N 4-[(4-anilino-5-sulfonaphthalen-1-yl)diazenyl]-5-hydroxynaphthalene-2,7-disulfonic acid Chemical compound C=12C(O)=CC(S(O)(=O)=O)=CC2=CC(S(O)(=O)=O)=CC=1N=NC(C1=CC=CC(=C11)S(O)(=O)=O)=CC=C1NC1=CC=CC=C1 QFVHZQCOUORWEI-UHFFFAOYSA-N 0.000 description 1
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- TTXMOJWKNRJWQJ-FXQIFTODSA-N Ala-Arg-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N TTXMOJWKNRJWQJ-FXQIFTODSA-N 0.000 description 1
- WCBVQNZTOKJWJS-ACZMJKKPSA-N Ala-Cys-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(O)=O WCBVQNZTOKJWJS-ACZMJKKPSA-N 0.000 description 1
- OYJCVIGKMXUVKB-GARJFASQSA-N Ala-Leu-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N OYJCVIGKMXUVKB-GARJFASQSA-N 0.000 description 1
- KQESEZXHYOUIIM-CQDKDKBSSA-N Ala-Lys-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KQESEZXHYOUIIM-CQDKDKBSSA-N 0.000 description 1
- IORKCNUBHNIMKY-CIUDSAMLSA-N Ala-Pro-Glu Chemical compound C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O IORKCNUBHNIMKY-CIUDSAMLSA-N 0.000 description 1
- RTZCUEHYUQZIDE-WHFBIAKZSA-N Ala-Ser-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O RTZCUEHYUQZIDE-WHFBIAKZSA-N 0.000 description 1
- ARHJJAAWNWOACN-FXQIFTODSA-N Ala-Ser-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O ARHJJAAWNWOACN-FXQIFTODSA-N 0.000 description 1
- CREYEAPXISDKSB-FQPOAREZSA-N Ala-Thr-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CREYEAPXISDKSB-FQPOAREZSA-N 0.000 description 1
- 206010061424 Anal cancer Diseases 0.000 description 1
- 208000007860 Anus Neoplasms Diseases 0.000 description 1
- 108010039627 Aprotinin Proteins 0.000 description 1
- AUFHLLPVPSMEOG-YUMQZZPRSA-N Arg-Gly-Glu Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O AUFHLLPVPSMEOG-YUMQZZPRSA-N 0.000 description 1
- LVMUGODRNHFGRA-AVGNSLFASA-N Arg-Leu-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O LVMUGODRNHFGRA-AVGNSLFASA-N 0.000 description 1
- AOHKLEBWKMKITA-IHRRRGAJSA-N Arg-Phe-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AOHKLEBWKMKITA-IHRRRGAJSA-N 0.000 description 1
- ADPACBMPYWJJCE-FXQIFTODSA-N Arg-Ser-Asp Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O ADPACBMPYWJJCE-FXQIFTODSA-N 0.000 description 1
- ICRHGPYYXMWHIE-LPEHRKFASA-N Arg-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O ICRHGPYYXMWHIE-LPEHRKFASA-N 0.000 description 1
- XRNXPIGJPQHCPC-RCWTZXSCSA-N Arg-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CCCNC(N)=N)[C@@H](C)O)C(O)=O XRNXPIGJPQHCPC-RCWTZXSCSA-N 0.000 description 1
- SLKLLQWZQHXYSV-CIUDSAMLSA-N Asn-Ala-Lys Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O SLKLLQWZQHXYSV-CIUDSAMLSA-N 0.000 description 1
- NVGWESORMHFISY-SRVKXCTJSA-N Asn-Asn-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O NVGWESORMHFISY-SRVKXCTJSA-N 0.000 description 1
- BVLIJXXSXBUGEC-SRVKXCTJSA-N Asn-Asn-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BVLIJXXSXBUGEC-SRVKXCTJSA-N 0.000 description 1
- RCFGLXMZDYNRSC-CIUDSAMLSA-N Asn-Lys-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O RCFGLXMZDYNRSC-CIUDSAMLSA-N 0.000 description 1
- SUIJFTJDTJKSRK-IHRRRGAJSA-N Asn-Pro-Tyr Chemical compound NC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 SUIJFTJDTJKSRK-IHRRRGAJSA-N 0.000 description 1
- MKJBPDLENBUHQU-CIUDSAMLSA-N Asn-Ser-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O MKJBPDLENBUHQU-CIUDSAMLSA-N 0.000 description 1
- WLVLIYYBPPONRJ-GCJQMDKQSA-N Asn-Thr-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O WLVLIYYBPPONRJ-GCJQMDKQSA-N 0.000 description 1
- MJIJBEYEHBKTIM-BYULHYEWSA-N Asn-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N MJIJBEYEHBKTIM-BYULHYEWSA-N 0.000 description 1
- KBQOUDLMWYWXNP-YDHLFZDLSA-N Asn-Val-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CC(=O)N)N KBQOUDLMWYWXNP-YDHLFZDLSA-N 0.000 description 1
- DBWYWXNMZZYIRY-LPEHRKFASA-N Asp-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(=O)O)N)C(=O)O DBWYWXNMZZYIRY-LPEHRKFASA-N 0.000 description 1
- XYBJLTKSGFBLCS-QXEWZRGKSA-N Asp-Arg-Val Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)CC(O)=O XYBJLTKSGFBLCS-QXEWZRGKSA-N 0.000 description 1
- QXHVOUSPVAWEMX-ZLUOBGJFSA-N Asp-Asp-Ser Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O QXHVOUSPVAWEMX-ZLUOBGJFSA-N 0.000 description 1
- VAWNQIGQPUOPQW-ACZMJKKPSA-N Asp-Glu-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O VAWNQIGQPUOPQW-ACZMJKKPSA-N 0.000 description 1
- WSGVTKZFVJSJOG-RCOVLWMOSA-N Asp-Gly-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O WSGVTKZFVJSJOG-RCOVLWMOSA-N 0.000 description 1
- DPNWSMBUYCLEDG-CIUDSAMLSA-N Asp-Lys-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O DPNWSMBUYCLEDG-CIUDSAMLSA-N 0.000 description 1
- DONWIPDSZZJHHK-HJGDQZAQSA-N Asp-Lys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(=O)O)N)O DONWIPDSZZJHHK-HJGDQZAQSA-N 0.000 description 1
- AHWRSSLYSGLBGD-CIUDSAMLSA-N Asp-Pro-Glu Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O AHWRSSLYSGLBGD-CIUDSAMLSA-N 0.000 description 1
- MGSVBZIBCCKGCY-ZLUOBGJFSA-N Asp-Ser-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MGSVBZIBCCKGCY-ZLUOBGJFSA-N 0.000 description 1
- KNDCWFXCFKSEBM-AVGNSLFASA-N Asp-Tyr-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O KNDCWFXCFKSEBM-AVGNSLFASA-N 0.000 description 1
- ALMIMUZAWTUNIO-BZSNNMDCSA-N Asp-Tyr-Tyr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ALMIMUZAWTUNIO-BZSNNMDCSA-N 0.000 description 1
- 206010003571 Astrocytoma Diseases 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 239000005711 Benzoic acid Substances 0.000 description 1
- 206010005003 Bladder cancer Diseases 0.000 description 1
- 206010005949 Bone cancer Diseases 0.000 description 1
- 208000018084 Bone neoplasm Diseases 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 206010006143 Brain stem glioma Diseases 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- 201000009030 Carcinoma Diseases 0.000 description 1
- 206010008342 Cervix carcinoma Diseases 0.000 description 1
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 1
- 108091026890 Coding region Proteins 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 241000699802 Cricetulus griseus Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- DCJNIJAWIRPPBB-CIUDSAMLSA-N Cys-Ala-Lys Chemical compound C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CS)N DCJNIJAWIRPPBB-CIUDSAMLSA-N 0.000 description 1
- GMXSSZUVDNPRMA-FXQIFTODSA-N Cys-Arg-Asp Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O GMXSSZUVDNPRMA-FXQIFTODSA-N 0.000 description 1
- WVLZTXGTNGHPBO-SRVKXCTJSA-N Cys-Leu-Leu Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O WVLZTXGTNGHPBO-SRVKXCTJSA-N 0.000 description 1
- SMEYEQDCCBHTEF-FXQIFTODSA-N Cys-Pro-Ala Chemical compound [H]N[C@@H](CS)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O SMEYEQDCCBHTEF-FXQIFTODSA-N 0.000 description 1
- ALTQTAKGRFLRLR-GUBZILKMSA-N Cys-Val-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CS)N ALTQTAKGRFLRLR-GUBZILKMSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- KDXKERNSBIXSRK-RXMQYKEDSA-N D-lysine Chemical compound NCCCC[C@@H](N)C(O)=O KDXKERNSBIXSRK-RXMQYKEDSA-N 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 206010014759 Endometrial neoplasm Diseases 0.000 description 1
- 206010014967 Ependymoma Diseases 0.000 description 1
- 208000000461 Esophageal Neoplasms Diseases 0.000 description 1
- 208000006168 Ewing Sarcoma Diseases 0.000 description 1
- 201000001342 Fallopian tube cancer Diseases 0.000 description 1
- 208000013452 Fallopian tube neoplasm Diseases 0.000 description 1
- 239000012739 FreeStyle 293 Expression medium Substances 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 201000010915 Glioblastoma multiforme Diseases 0.000 description 1
- ITYRYNUZHPNCIK-GUBZILKMSA-N Glu-Ala-Leu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O ITYRYNUZHPNCIK-GUBZILKMSA-N 0.000 description 1
- PAQUJCSYVIBPLC-AVGNSLFASA-N Glu-Asp-Phe Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 PAQUJCSYVIBPLC-AVGNSLFASA-N 0.000 description 1
- CKOFNWCLWRYUHK-XHNCKOQMSA-N Glu-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCC(=O)O)N)C(=O)O CKOFNWCLWRYUHK-XHNCKOQMSA-N 0.000 description 1
- KOSRFJWDECSPRO-WDSKDSINSA-N Glu-Glu Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(O)=O KOSRFJWDECSPRO-WDSKDSINSA-N 0.000 description 1
- FBEJIDRSQCGFJI-GUBZILKMSA-N Glu-Leu-Ser Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O FBEJIDRSQCGFJI-GUBZILKMSA-N 0.000 description 1
- NJCALAAIGREHDR-WDCWCFNPSA-N Glu-Leu-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NJCALAAIGREHDR-WDCWCFNPSA-N 0.000 description 1
- BFEZQZKEPRKKHV-SRVKXCTJSA-N Glu-Pro-Lys Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCC(=O)O)N)C(=O)N[C@@H](CCCCN)C(=O)O BFEZQZKEPRKKHV-SRVKXCTJSA-N 0.000 description 1
- NNQDRRUXFJYCCJ-NHCYSSNCSA-N Glu-Pro-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(O)=O NNQDRRUXFJYCCJ-NHCYSSNCSA-N 0.000 description 1
- SITLTJHOQZFJGG-XPUUQOCRSA-N Glu-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H](N)CCC(O)=O SITLTJHOQZFJGG-XPUUQOCRSA-N 0.000 description 1
- ZALGPUWUVHOGAE-GVXVVHGQSA-N Glu-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCC(=O)O)N ZALGPUWUVHOGAE-GVXVVHGQSA-N 0.000 description 1
- WGYHAAXZWPEBDQ-IFFSRLJSSA-N Glu-Val-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WGYHAAXZWPEBDQ-IFFSRLJSSA-N 0.000 description 1
- VSVZIEVNUYDAFR-YUMQZZPRSA-N Gly-Ala-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN VSVZIEVNUYDAFR-YUMQZZPRSA-N 0.000 description 1
- MZZSCEANQDPJER-ONGXEEELSA-N Gly-Ala-Phe Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 MZZSCEANQDPJER-ONGXEEELSA-N 0.000 description 1
- CIMULJZTTOBOPN-WHFBIAKZSA-N Gly-Asn-Asn Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CIMULJZTTOBOPN-WHFBIAKZSA-N 0.000 description 1
- GRIRDMVMJJDZKV-RCOVLWMOSA-N Gly-Asn-Val Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O GRIRDMVMJJDZKV-RCOVLWMOSA-N 0.000 description 1
- YWAQATDNEKZFFK-BYPYZUCNSA-N Gly-Gly-Ser Chemical compound NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O YWAQATDNEKZFFK-BYPYZUCNSA-N 0.000 description 1
- ICUTTWWCDIIIEE-BQBZGAKWSA-N Gly-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)CN ICUTTWWCDIIIEE-BQBZGAKWSA-N 0.000 description 1
- HAOUOFNNJJLVNS-BQBZGAKWSA-N Gly-Pro-Ser Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O HAOUOFNNJJLVNS-BQBZGAKWSA-N 0.000 description 1
- POJJAZJHBGXEGM-YUMQZZPRSA-N Gly-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)CN POJJAZJHBGXEGM-YUMQZZPRSA-N 0.000 description 1
- NVTPVQLIZCOJFK-FOHZUACHSA-N Gly-Thr-Asp Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O NVTPVQLIZCOJFK-FOHZUACHSA-N 0.000 description 1
- JQFILXICXLDTRR-FBCQKBJTSA-N Gly-Thr-Gly Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)NCC(O)=O JQFILXICXLDTRR-FBCQKBJTSA-N 0.000 description 1
- XHVONGZZVUUORG-WEDXCCLWSA-N Gly-Thr-Lys Chemical compound NCC(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CCCCN XHVONGZZVUUORG-WEDXCCLWSA-N 0.000 description 1
- NGBGZCUWFVVJKC-IRXDYDNUSA-N Gly-Tyr-Tyr Chemical compound C([C@H](NC(=O)CN)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 NGBGZCUWFVVJKC-IRXDYDNUSA-N 0.000 description 1
- FULZDMOZUZKGQU-ONGXEEELSA-N Gly-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)CN FULZDMOZUZKGQU-ONGXEEELSA-N 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- MAABHGXCIBEYQR-XVYDVKMFSA-N His-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N MAABHGXCIBEYQR-XVYDVKMFSA-N 0.000 description 1
- WMKXFMUJRCEGRP-SRVKXCTJSA-N His-Asn-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N WMKXFMUJRCEGRP-SRVKXCTJSA-N 0.000 description 1
- HIAHVKLTHNOENC-HGNGGELXSA-N His-Glu-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O HIAHVKLTHNOENC-HGNGGELXSA-N 0.000 description 1
- TTYKEFZRLKQTHH-MELADBBJSA-N His-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O TTYKEFZRLKQTHH-MELADBBJSA-N 0.000 description 1
- ZHHLTWUOWXHVQJ-YUMQZZPRSA-N His-Ser-Gly Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZHHLTWUOWXHVQJ-YUMQZZPRSA-N 0.000 description 1
- HZWWOGWOBQBETJ-CUJWVEQBSA-N His-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N)O HZWWOGWOBQBETJ-CUJWVEQBSA-N 0.000 description 1
- ALPXXNRQBMRCPZ-MEYUZBJRSA-N His-Thr-Phe Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O ALPXXNRQBMRCPZ-MEYUZBJRSA-N 0.000 description 1
- CSTDQOOBZBAJKE-BWAGICSOSA-N His-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CC2=CN=CN2)N)O CSTDQOOBZBAJKE-BWAGICSOSA-N 0.000 description 1
- 101000958041 Homo sapiens Musculin Proteins 0.000 description 1
- 102000003839 Human Proteins Human genes 0.000 description 1
- 108090000144 Human Proteins Proteins 0.000 description 1
- 102000009786 Immunoglobulin Constant Regions Human genes 0.000 description 1
- 108010009817 Immunoglobulin Constant Regions Proteins 0.000 description 1
- 102000006496 Immunoglobulin Heavy Chains Human genes 0.000 description 1
- 108010019476 Immunoglobulin Heavy Chains Proteins 0.000 description 1
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 1
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 1
- 206010061252 Intraocular melanoma Diseases 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- 208000008839 Kidney Neoplasms Diseases 0.000 description 1
- QLROSWPKSBORFJ-BQBZGAKWSA-N L-Prolyl-L-glutamic acid Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1 QLROSWPKSBORFJ-BQBZGAKWSA-N 0.000 description 1
- KZSNJWFQEVHDMF-BYPYZUCNSA-N L-valine Chemical compound CC(C)[C@H](N)C(O)=O KZSNJWFQEVHDMF-BYPYZUCNSA-N 0.000 description 1
- USTCFDAQCLDPBD-XIRDDKMYSA-N Leu-Asn-Trp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N USTCFDAQCLDPBD-XIRDDKMYSA-N 0.000 description 1
- QJUWBDPGGYVRHY-YUMQZZPRSA-N Leu-Gly-Cys Chemical compound CC(C)C[C@@H](C(=O)NCC(=O)N[C@@H](CS)C(=O)O)N QJUWBDPGGYVRHY-YUMQZZPRSA-N 0.000 description 1
- VWHGTYCRDRBSFI-ZETCQYMHSA-N Leu-Gly-Gly Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)NCC(O)=O VWHGTYCRDRBSFI-ZETCQYMHSA-N 0.000 description 1
- HYMLKESRWLZDBR-WEDXCCLWSA-N Leu-Gly-Thr Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O HYMLKESRWLZDBR-WEDXCCLWSA-N 0.000 description 1
- XWOBNBRUDDUEEY-UWVGGRQHSA-N Leu-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(O)=O)CC1=CNC=N1 XWOBNBRUDDUEEY-UWVGGRQHSA-N 0.000 description 1
- BTNXKBVLWJBTNR-SRVKXCTJSA-N Leu-His-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O BTNXKBVLWJBTNR-SRVKXCTJSA-N 0.000 description 1
- YOKVEHGYYQEQOP-QWRGUYRKSA-N Leu-Leu-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YOKVEHGYYQEQOP-QWRGUYRKSA-N 0.000 description 1
- WMIOEVKKYIMVKI-DCAQKATOSA-N Leu-Pro-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O WMIOEVKKYIMVKI-DCAQKATOSA-N 0.000 description 1
- QWWPYKKLXWOITQ-VOAKCMCISA-N Leu-Thr-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(C)C QWWPYKKLXWOITQ-VOAKCMCISA-N 0.000 description 1
- AIQWYVFNBNNOLU-RHYQMDGZSA-N Leu-Thr-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O AIQWYVFNBNNOLU-RHYQMDGZSA-N 0.000 description 1
- YQFZRHYZLARWDY-IHRRRGAJSA-N Leu-Val-Lys Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN YQFZRHYZLARWDY-IHRRRGAJSA-N 0.000 description 1
- 208000028018 Lymphocytic leukaemia Diseases 0.000 description 1
- 206010025323 Lymphomas Diseases 0.000 description 1
- IXHKPDJKKCUKHS-GARJFASQSA-N Lys-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N IXHKPDJKKCUKHS-GARJFASQSA-N 0.000 description 1
- UWKNTTJNVSYXPC-CIUDSAMLSA-N Lys-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN UWKNTTJNVSYXPC-CIUDSAMLSA-N 0.000 description 1
- CLBGMWIYPYAZPR-AVGNSLFASA-N Lys-Arg-Arg Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O CLBGMWIYPYAZPR-AVGNSLFASA-N 0.000 description 1
- KWUKZRFFKPLUPE-HJGDQZAQSA-N Lys-Asp-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWUKZRFFKPLUPE-HJGDQZAQSA-N 0.000 description 1
- FGMHXLULNHTPID-KKUMJFAQSA-N Lys-His-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CN=CN1 FGMHXLULNHTPID-KKUMJFAQSA-N 0.000 description 1
- WQDKIVRHTQYJSN-DCAQKATOSA-N Lys-Ser-Arg Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N WQDKIVRHTQYJSN-DCAQKATOSA-N 0.000 description 1
- DIBZLYZXTSVGLN-CIUDSAMLSA-N Lys-Ser-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O DIBZLYZXTSVGLN-CIUDSAMLSA-N 0.000 description 1
- IEVXCWPVBYCJRZ-IXOXFDKPSA-N Lys-Thr-His Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 IEVXCWPVBYCJRZ-IXOXFDKPSA-N 0.000 description 1
- YKBSXQFZWFXFIB-VOAKCMCISA-N Lys-Thr-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCCN)C(O)=O YKBSXQFZWFXFIB-VOAKCMCISA-N 0.000 description 1
- CAVRAQIDHUPECU-UVOCVTCTSA-N Lys-Thr-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O CAVRAQIDHUPECU-UVOCVTCTSA-N 0.000 description 1
- YQAIUOWPSUOINN-IUCAKERBSA-N Lys-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H](N)CCCCN YQAIUOWPSUOINN-IUCAKERBSA-N 0.000 description 1
- QLFAPXUXEBAWEK-NHCYSSNCSA-N Lys-Val-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O QLFAPXUXEBAWEK-NHCYSSNCSA-N 0.000 description 1
- UGCIQUYEJIEHKX-GVXVVHGQSA-N Lys-Val-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O UGCIQUYEJIEHKX-GVXVVHGQSA-N 0.000 description 1
- DRRXXZBXDMLGFC-IHRRRGAJSA-N Lys-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCCN DRRXXZBXDMLGFC-IHRRRGAJSA-N 0.000 description 1
- 208000032271 Malignant tumor of penis Diseases 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 208000000172 Medulloblastoma Diseases 0.000 description 1
- 206010027406 Mesothelioma Diseases 0.000 description 1
- CAODKDAPYGUMLK-FXQIFTODSA-N Met-Asn-Ser Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O CAODKDAPYGUMLK-FXQIFTODSA-N 0.000 description 1
- SJDQOYTYNGZZJX-SRVKXCTJSA-N Met-Glu-Leu Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O SJDQOYTYNGZZJX-SRVKXCTJSA-N 0.000 description 1
- ABHVWYPPHDYFNY-WDSOQIARSA-N Met-His-Trp Chemical compound C([C@H](NC(=O)[C@@H](N)CCSC)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)C1=CN=CN1 ABHVWYPPHDYFNY-WDSOQIARSA-N 0.000 description 1
- KAKJTZWHIUWTTD-VQVTYTSYSA-N Met-Thr Chemical compound CSCC[C@H]([NH3+])C(=O)N[C@@H]([C@@H](C)O)C([O-])=O KAKJTZWHIUWTTD-VQVTYTSYSA-N 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- AUEJLPRZGVVDNU-UHFFFAOYSA-N N-L-tyrosyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CC1=CC=C(O)C=C1 AUEJLPRZGVVDNU-UHFFFAOYSA-N 0.000 description 1
- 230000004988 N-glycosylation Effects 0.000 description 1
- CHJJGSNFBQVOTG-UHFFFAOYSA-N N-methyl-guanidine Natural products CNC(N)=N CHJJGSNFBQVOTG-UHFFFAOYSA-N 0.000 description 1
- 108091007491 NSP3 Papain-like protease domains Proteins 0.000 description 1
- BQVUABVGYYSDCJ-UHFFFAOYSA-N Nalpha-L-Leucyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)CC(C)C)C(O)=O)=CNC2=C1 BQVUABVGYYSDCJ-UHFFFAOYSA-N 0.000 description 1
- 229930193140 Neomycin Natural products 0.000 description 1
- 102000019040 Nuclear Antigens Human genes 0.000 description 1
- 108010051791 Nuclear Antigens Proteins 0.000 description 1
- 108091005461 Nucleic proteins Proteins 0.000 description 1
- DMGNFLJBACZMRM-UHFFFAOYSA-N O[P] Chemical compound O[P] DMGNFLJBACZMRM-UHFFFAOYSA-N 0.000 description 1
- 206010030155 Oesophageal carcinoma Diseases 0.000 description 1
- 206010033128 Ovarian cancer Diseases 0.000 description 1
- 206010061535 Ovarian neoplasm Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 208000000821 Parathyroid Neoplasms Diseases 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 208000002471 Penile Neoplasms Diseases 0.000 description 1
- 206010034299 Penile cancer Diseases 0.000 description 1
- 102000000447 Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase Human genes 0.000 description 1
- 108010055817 Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase Proteins 0.000 description 1
- JOXIIFVCSATTDH-IHPCNDPISA-N Phe-Asn-Trp Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC2=CNC3=CC=CC=C32)C(=O)O)N JOXIIFVCSATTDH-IHPCNDPISA-N 0.000 description 1
- MSHZERMPZKCODG-ACRUOGEOSA-N Phe-Leu-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 MSHZERMPZKCODG-ACRUOGEOSA-N 0.000 description 1
- KNYPNEYICHHLQL-ACRUOGEOSA-N Phe-Leu-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 KNYPNEYICHHLQL-ACRUOGEOSA-N 0.000 description 1
- DMEYUTSDVRCWRS-ULQDDVLXSA-N Phe-Lys-Arg Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=CC=C1 DMEYUTSDVRCWRS-ULQDDVLXSA-N 0.000 description 1
- AXIOGMQCDYVTNY-ACRUOGEOSA-N Phe-Phe-Leu Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 AXIOGMQCDYVTNY-ACRUOGEOSA-N 0.000 description 1
- WWPAHTZOWURIMR-ULQDDVLXSA-N Phe-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=CC=C1 WWPAHTZOWURIMR-ULQDDVLXSA-N 0.000 description 1
- IIEOLPMQYRBZCN-SRVKXCTJSA-N Phe-Ser-Cys Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O IIEOLPMQYRBZCN-SRVKXCTJSA-N 0.000 description 1
- BPCLGWHVPVTTFM-QWRGUYRKSA-N Phe-Ser-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)NCC(O)=O BPCLGWHVPVTTFM-QWRGUYRKSA-N 0.000 description 1
- BSKMOCNNLNDIMU-CDMKHQONSA-N Phe-Thr-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O BSKMOCNNLNDIMU-CDMKHQONSA-N 0.000 description 1
- KLYYKKGCPOGDPE-OEAJRASXSA-N Phe-Thr-Leu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O KLYYKKGCPOGDPE-OEAJRASXSA-N 0.000 description 1
- 208000007913 Pituitary Neoplasms Diseases 0.000 description 1
- 201000005746 Pituitary adenoma Diseases 0.000 description 1
- 206010061538 Pituitary tumour benign Diseases 0.000 description 1
- OOLOTUZJUBOMAX-GUBZILKMSA-N Pro-Ala-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O OOLOTUZJUBOMAX-GUBZILKMSA-N 0.000 description 1
- TXPUNZXZDVJUJQ-LPEHRKFASA-N Pro-Asn-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC(=O)N)C(=O)N2CCC[C@@H]2C(=O)O TXPUNZXZDVJUJQ-LPEHRKFASA-N 0.000 description 1
- PULPZRAHVFBVTO-DCAQKATOSA-N Pro-Glu-Arg Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O PULPZRAHVFBVTO-DCAQKATOSA-N 0.000 description 1
- KIPIKSXPPLABPN-CIUDSAMLSA-N Pro-Glu-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H]1CCCN1 KIPIKSXPPLABPN-CIUDSAMLSA-N 0.000 description 1
- NXEYSLRNNPWCRN-SRVKXCTJSA-N Pro-Glu-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NXEYSLRNNPWCRN-SRVKXCTJSA-N 0.000 description 1
- CLNJSLSHKJECME-BQBZGAKWSA-N Pro-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H]1CCCN1 CLNJSLSHKJECME-BQBZGAKWSA-N 0.000 description 1
- FKLSMYYLJHYPHH-UWVGGRQHSA-N Pro-Gly-Leu Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O FKLSMYYLJHYPHH-UWVGGRQHSA-N 0.000 description 1
- ZLXKLMHAMDENIO-DCAQKATOSA-N Pro-Lys-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLXKLMHAMDENIO-DCAQKATOSA-N 0.000 description 1
- ULWBBFKQBDNGOY-RWMBFGLXSA-N Pro-Lys-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCCCN)C(=O)N2CCC[C@@H]2C(=O)O ULWBBFKQBDNGOY-RWMBFGLXSA-N 0.000 description 1
- HWLKHNDRXWTFTN-GUBZILKMSA-N Pro-Pro-Cys Chemical compound C1C[C@H](NC1)C(=O)N2CCC[C@H]2C(=O)N[C@@H](CS)C(=O)O HWLKHNDRXWTFTN-GUBZILKMSA-N 0.000 description 1
- FDMKYQQYJKYCLV-GUBZILKMSA-N Pro-Pro-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 FDMKYQQYJKYCLV-GUBZILKMSA-N 0.000 description 1
- KBUAPZAZPWNYSW-SRVKXCTJSA-N Pro-Pro-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 KBUAPZAZPWNYSW-SRVKXCTJSA-N 0.000 description 1
- 206010060862 Prostate cancer Diseases 0.000 description 1
- 208000000236 Prostatic Neoplasms Diseases 0.000 description 1
- 208000015634 Rectal Neoplasms Diseases 0.000 description 1
- 206010038389 Renal cancer Diseases 0.000 description 1
- 208000006265 Renal cell carcinoma Diseases 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- 206010039491 Sarcoma Diseases 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 1
- QEDMOZUJTGEIBF-FXQIFTODSA-N Ser-Arg-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O QEDMOZUJTGEIBF-FXQIFTODSA-N 0.000 description 1
- WDXYVIIVDIDOSX-DCAQKATOSA-N Ser-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CCCN=C(N)N WDXYVIIVDIDOSX-DCAQKATOSA-N 0.000 description 1
- VGNYHOBZJKWRGI-CIUDSAMLSA-N Ser-Asn-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CO VGNYHOBZJKWRGI-CIUDSAMLSA-N 0.000 description 1
- UGJRQLURDVGULT-LKXGYXEUSA-N Ser-Asn-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UGJRQLURDVGULT-LKXGYXEUSA-N 0.000 description 1
- ICHZYBVODUVUKN-SRVKXCTJSA-N Ser-Asn-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ICHZYBVODUVUKN-SRVKXCTJSA-N 0.000 description 1
- VBKBDLMWICBSCY-IMJSIDKUSA-N Ser-Asp Chemical compound OC[C@H](N)C(=O)N[C@H](C(O)=O)CC(O)=O VBKBDLMWICBSCY-IMJSIDKUSA-N 0.000 description 1
- MESDJCNHLZBMEP-ZLUOBGJFSA-N Ser-Asp-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MESDJCNHLZBMEP-ZLUOBGJFSA-N 0.000 description 1
- KCFKKAQKRZBWJB-ZLUOBGJFSA-N Ser-Cys-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O KCFKKAQKRZBWJB-ZLUOBGJFSA-N 0.000 description 1
- KNCJWSPMTFFJII-ZLUOBGJFSA-N Ser-Cys-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(O)=O)C(O)=O KNCJWSPMTFFJII-ZLUOBGJFSA-N 0.000 description 1
- KMWFXJCGRXBQAC-CIUDSAMLSA-N Ser-Cys-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CO)N KMWFXJCGRXBQAC-CIUDSAMLSA-N 0.000 description 1
- MOVJSUIKUNCVMG-ZLUOBGJFSA-N Ser-Cys-Ser Chemical compound C([C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(=O)O)N)O MOVJSUIKUNCVMG-ZLUOBGJFSA-N 0.000 description 1
- UIGMAMGZOJVTDN-WHFBIAKZSA-N Ser-Gly-Ser Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O UIGMAMGZOJVTDN-WHFBIAKZSA-N 0.000 description 1
- HMRAQFJFTOLDKW-GUBZILKMSA-N Ser-His-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(O)=O HMRAQFJFTOLDKW-GUBZILKMSA-N 0.000 description 1
- XUDRHBPSPAPDJP-SRVKXCTJSA-N Ser-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CO XUDRHBPSPAPDJP-SRVKXCTJSA-N 0.000 description 1
- GDUZTEQRAOXYJS-SRVKXCTJSA-N Ser-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CO)N GDUZTEQRAOXYJS-SRVKXCTJSA-N 0.000 description 1
- AZWNCEBQZXELEZ-FXQIFTODSA-N Ser-Pro-Ser Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O AZWNCEBQZXELEZ-FXQIFTODSA-N 0.000 description 1
- NADLKBTYNKUJEP-KATARQTJSA-N Ser-Thr-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NADLKBTYNKUJEP-KATARQTJSA-N 0.000 description 1
- SNXUIBACCONSOH-BWBBJGPYSA-N Ser-Thr-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CO)C(O)=O SNXUIBACCONSOH-BWBBJGPYSA-N 0.000 description 1
- IAOHCSQDQDWRQU-GUBZILKMSA-N Ser-Val-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O IAOHCSQDQDWRQU-GUBZILKMSA-N 0.000 description 1
- LGIMRDKGABDMBN-DCAQKATOSA-N Ser-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CO)N LGIMRDKGABDMBN-DCAQKATOSA-N 0.000 description 1
- RCOUFINCYASMDN-GUBZILKMSA-N Ser-Val-Met Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCSC)C(O)=O RCOUFINCYASMDN-GUBZILKMSA-N 0.000 description 1
- 201000004283 Shwachman-Diamond syndrome Diseases 0.000 description 1
- 208000000453 Skin Neoplasms Diseases 0.000 description 1
- 208000021712 Soft tissue sarcoma Diseases 0.000 description 1
- PZBFGYYEXUXCOF-UHFFFAOYSA-N TCEP Chemical compound OC(=O)CCP(CCC(O)=O)CCC(O)=O PZBFGYYEXUXCOF-UHFFFAOYSA-N 0.000 description 1
- IGROJMCBGRFRGI-YTLHQDLWSA-N Thr-Ala-Ala Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O IGROJMCBGRFRGI-YTLHQDLWSA-N 0.000 description 1
- MQCPGOZXFSYJPS-KZVJFYERSA-N Thr-Ala-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MQCPGOZXFSYJPS-KZVJFYERSA-N 0.000 description 1
- DWYAUVCQDTZIJI-VZFHVOOUSA-N Thr-Ala-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O DWYAUVCQDTZIJI-VZFHVOOUSA-N 0.000 description 1
- XSLXHSYIVPGEER-KZVJFYERSA-N Thr-Ala-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(O)=O XSLXHSYIVPGEER-KZVJFYERSA-N 0.000 description 1
- LYGKYFKSZTUXGZ-ZDLURKLDSA-N Thr-Cys-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)NCC(O)=O LYGKYFKSZTUXGZ-ZDLURKLDSA-N 0.000 description 1
- KWQBJOUOSNJDRR-XAVMHZPKSA-N Thr-Cys-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)N1CCC[C@@H]1C(=O)O)N)O KWQBJOUOSNJDRR-XAVMHZPKSA-N 0.000 description 1
- MMTOHPRBJKEZHT-BWBBJGPYSA-N Thr-Cys-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(O)=O MMTOHPRBJKEZHT-BWBBJGPYSA-N 0.000 description 1
- DSLHSTIUAPKERR-XGEHTFHBSA-N Thr-Cys-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(O)=O DSLHSTIUAPKERR-XGEHTFHBSA-N 0.000 description 1
- BECPPKYKPSRKCP-ZDLURKLDSA-N Thr-Glu Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(O)=O)CCC(O)=O BECPPKYKPSRKCP-ZDLURKLDSA-N 0.000 description 1
- JQAWYCUUFIMTHE-WLTAIBSBSA-N Thr-Gly-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JQAWYCUUFIMTHE-WLTAIBSBSA-N 0.000 description 1
- FIFDDJFLNVAVMS-RHYQMDGZSA-N Thr-Leu-Met Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(O)=O FIFDDJFLNVAVMS-RHYQMDGZSA-N 0.000 description 1
- VRUFCJZQDACGLH-UVOCVTCTSA-N Thr-Leu-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VRUFCJZQDACGLH-UVOCVTCTSA-N 0.000 description 1
- BDGBHYCAZJPLHX-HJGDQZAQSA-N Thr-Lys-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O BDGBHYCAZJPLHX-HJGDQZAQSA-N 0.000 description 1
- JLNMFGCJODTXDH-WEDXCCLWSA-N Thr-Lys-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O JLNMFGCJODTXDH-WEDXCCLWSA-N 0.000 description 1
- BIBYEFRASCNLAA-CDMKHQONSA-N Thr-Phe-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=CC=C1 BIBYEFRASCNLAA-CDMKHQONSA-N 0.000 description 1
- MROIJTGJGIDEEJ-RCWTZXSCSA-N Thr-Pro-Pro Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 MROIJTGJGIDEEJ-RCWTZXSCSA-N 0.000 description 1
- FYBFTPLPAXZBOY-KKHAAJSZSA-N Thr-Val-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O FYBFTPLPAXZBOY-KKHAAJSZSA-N 0.000 description 1
- BKVICMPZWRNWOC-RHYQMDGZSA-N Thr-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)[C@@H](C)O BKVICMPZWRNWOC-RHYQMDGZSA-N 0.000 description 1
- QNXZCKMXHPULME-ZNSHCXBVSA-N Thr-Val-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N)O QNXZCKMXHPULME-ZNSHCXBVSA-N 0.000 description 1
- MNYNCKZAEIAONY-XGEHTFHBSA-N Thr-Val-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O MNYNCKZAEIAONY-XGEHTFHBSA-N 0.000 description 1
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 description 1
- 239000004473 Threonine Substances 0.000 description 1
- 208000024770 Thyroid neoplasm Diseases 0.000 description 1
- 235000011941 Tilia x europaea Nutrition 0.000 description 1
- UKINEYBQXPMOJO-UBHSHLNASA-N Trp-Asn-Ser Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N UKINEYBQXPMOJO-UBHSHLNASA-N 0.000 description 1
- BSSJIVIFAJKLEK-XIRDDKMYSA-N Trp-Cys-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N BSSJIVIFAJKLEK-XIRDDKMYSA-N 0.000 description 1
- RERRMBXDSFMBQE-ZFWWWQNUSA-N Trp-Met-Gly Chemical compound CSCC[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N RERRMBXDSFMBQE-ZFWWWQNUSA-N 0.000 description 1
- SSSDKJMQMZTMJP-BVSLBCMMSA-N Trp-Tyr-Val Chemical compound C([C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)C1=CC=C(O)C=C1 SSSDKJMQMZTMJP-BVSLBCMMSA-N 0.000 description 1
- OLWFDNLLBWQWCP-STQMWFEESA-N Tyr-Gly-Met Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(=O)N[C@@H](CCSC)C(O)=O OLWFDNLLBWQWCP-STQMWFEESA-N 0.000 description 1
- SINRIKQYQJRGDQ-MEYUZBJRSA-N Tyr-Lys-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 SINRIKQYQJRGDQ-MEYUZBJRSA-N 0.000 description 1
- XYNFFTNEQDWZNY-ULQDDVLXSA-N Tyr-Met-His Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N XYNFFTNEQDWZNY-ULQDDVLXSA-N 0.000 description 1
- WURLIFOWSMBUAR-SLFFLAALSA-N Tyr-Phe-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)NC(=O)[C@H](CC3=CC=C(C=C3)O)N)C(=O)O WURLIFOWSMBUAR-SLFFLAALSA-N 0.000 description 1
- VBFVQTPETKJCQW-RPTUDFQQSA-N Tyr-Phe-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VBFVQTPETKJCQW-RPTUDFQQSA-N 0.000 description 1
- AUZADXNWQMBZOO-JYJNAYRXSA-N Tyr-Pro-Arg Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O)C1=CC=C(O)C=C1 AUZADXNWQMBZOO-JYJNAYRXSA-N 0.000 description 1
- YYLHVUCSTXXKBS-IHRRRGAJSA-N Tyr-Pro-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O YYLHVUCSTXXKBS-IHRRRGAJSA-N 0.000 description 1
- LUMQYLVYUIRHHU-YJRXYDGGSA-N Tyr-Ser-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LUMQYLVYUIRHHU-YJRXYDGGSA-N 0.000 description 1
- LDKDSFQSEUOCOO-RPTUDFQQSA-N Tyr-Thr-Phe Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O LDKDSFQSEUOCOO-RPTUDFQQSA-N 0.000 description 1
- OJCISMMNNUNNJA-BZSNNMDCSA-N Tyr-Tyr-Asp Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CC(O)=O)C(O)=O)C1=CC=C(O)C=C1 OJCISMMNNUNNJA-BZSNNMDCSA-N 0.000 description 1
- ANHVRCNNGJMJNG-BZSNNMDCSA-N Tyr-Tyr-Cys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CS)C(=O)O)N)O ANHVRCNNGJMJNG-BZSNNMDCSA-N 0.000 description 1
- NVJCMGGZHOJNBU-UFYCRDLUSA-N Tyr-Val-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N NVJCMGGZHOJNBU-UFYCRDLUSA-N 0.000 description 1
- 208000023915 Ureteral Neoplasms Diseases 0.000 description 1
- 206010046392 Ureteric cancer Diseases 0.000 description 1
- 206010046431 Urethral cancer Diseases 0.000 description 1
- 206010046458 Urethral neoplasms Diseases 0.000 description 1
- 208000007097 Urinary Bladder Neoplasms Diseases 0.000 description 1
- 208000006105 Uterine Cervical Neoplasms Diseases 0.000 description 1
- 208000002495 Uterine Neoplasms Diseases 0.000 description 1
- 201000005969 Uveal melanoma Diseases 0.000 description 1
- QHDXUYOYTPWCSK-RCOVLWMOSA-N Val-Asp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)NCC(=O)O)N QHDXUYOYTPWCSK-RCOVLWMOSA-N 0.000 description 1
- SCBITHMBEJNRHC-LSJOCFKGSA-N Val-Asp-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](C(C)C)C(=O)O)N SCBITHMBEJNRHC-LSJOCFKGSA-N 0.000 description 1
- IRLYZKKNBFPQBW-XGEHTFHBSA-N Val-Cys-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](C(C)C)N)O IRLYZKKNBFPQBW-XGEHTFHBSA-N 0.000 description 1
- XGJLNBNZNMVJRS-NRPADANISA-N Val-Glu-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O XGJLNBNZNMVJRS-NRPADANISA-N 0.000 description 1
- UEHRGZCNLSWGHK-DLOVCJGASA-N Val-Glu-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O UEHRGZCNLSWGHK-DLOVCJGASA-N 0.000 description 1
- XTDDIVQWDXMRJL-IHRRRGAJSA-N Val-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C(C)C)N XTDDIVQWDXMRJL-IHRRRGAJSA-N 0.000 description 1
- DIOSYUIWOQCXNR-ONGXEEELSA-N Val-Lys-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)NCC(O)=O DIOSYUIWOQCXNR-ONGXEEELSA-N 0.000 description 1
- HPANGHISDXDUQY-ULQDDVLXSA-N Val-Lys-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N HPANGHISDXDUQY-ULQDDVLXSA-N 0.000 description 1
- VPGCVZRRBYOGCD-AVGNSLFASA-N Val-Lys-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(O)=O VPGCVZRRBYOGCD-AVGNSLFASA-N 0.000 description 1
- SBJCTAZFSZXWSR-AVGNSLFASA-N Val-Met-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N SBJCTAZFSZXWSR-AVGNSLFASA-N 0.000 description 1
- HJSLDXZAZGFPDK-ULQDDVLXSA-N Val-Phe-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)[C@H](C(C)C)N HJSLDXZAZGFPDK-ULQDDVLXSA-N 0.000 description 1
- NSUUANXHLKKHQB-BZSNNMDCSA-N Val-Pro-Trp Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CNC2=CC=CC=C12 NSUUANXHLKKHQB-BZSNNMDCSA-N 0.000 description 1
- JQTYTBPCSOAZHI-FXQIFTODSA-N Val-Ser-Cys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CS)C(=O)O)N JQTYTBPCSOAZHI-FXQIFTODSA-N 0.000 description 1
- KRAHMIJVUPUOTQ-DCAQKATOSA-N Val-Ser-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N KRAHMIJVUPUOTQ-DCAQKATOSA-N 0.000 description 1
- QTPQHINADBYBNA-DCAQKATOSA-N Val-Ser-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCCN QTPQHINADBYBNA-DCAQKATOSA-N 0.000 description 1
- NZYNRRGJJVSSTJ-GUBZILKMSA-N Val-Ser-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NZYNRRGJJVSSTJ-GUBZILKMSA-N 0.000 description 1
- SUGRIIAOLCDLBD-ZOBUZTSGSA-N Val-Trp-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(=O)O)C(=O)O)N SUGRIIAOLCDLBD-ZOBUZTSGSA-N 0.000 description 1
- QHSSPPHOHJSTML-HOCLYGCPSA-N Val-Trp-Gly Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)NCC(=O)O)N QHSSPPHOHJSTML-HOCLYGCPSA-N 0.000 description 1
- BGTDGENDNWGMDQ-KJEVXHAQSA-N Val-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N)O BGTDGENDNWGMDQ-KJEVXHAQSA-N 0.000 description 1
- ZLNYBMWGPOKSLW-LSJOCFKGSA-N Val-Val-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O ZLNYBMWGPOKSLW-LSJOCFKGSA-N 0.000 description 1
- KZSNJWFQEVHDMF-UHFFFAOYSA-N Valine Natural products CC(C)C(N)C(O)=O KZSNJWFQEVHDMF-UHFFFAOYSA-N 0.000 description 1
- 206010047741 Vulval cancer Diseases 0.000 description 1
- 208000004354 Vulvar Neoplasms Diseases 0.000 description 1
- 239000002250 absorbent Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000002671 adjuvant Substances 0.000 description 1
- 201000005188 adrenal gland cancer Diseases 0.000 description 1
- 208000024447 adrenal gland neoplasm Diseases 0.000 description 1
- 238000005377 adsorption chromatography Methods 0.000 description 1
- 238000005054 agglomeration Methods 0.000 description 1
- 230000004520 agglutination Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 108010011559 alanylphenylalanine Proteins 0.000 description 1
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 201000011165 anus cancer Diseases 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 229960004405 aprotinin Drugs 0.000 description 1
- 239000012062 aqueous buffer Substances 0.000 description 1
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 1
- 108010068380 arginylarginine Proteins 0.000 description 1
- 125000003118 aryl group Chemical group 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 230000001588 bifunctional effect Effects 0.000 description 1
- 230000008033 biological extinction Effects 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 238000005251 capillar electrophoresis Methods 0.000 description 1
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 1
- 125000002057 carboxymethyl group Chemical group [H]OC(=O)C([H])([H])[*] 0.000 description 1
- 238000005341 cation exchange Methods 0.000 description 1
- 230000003915 cell function Effects 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 238000003570 cell viability assay Methods 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 201000010881 cervical cancer Diseases 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000013522 chelant Substances 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 238000005660 chlorination reaction Methods 0.000 description 1
- 208000006990 cholangiocarcinoma Diseases 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 230000002759 chromosomal effect Effects 0.000 description 1
- 210000000078 claw Anatomy 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 238000012761 co-transfection Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000004154 complement system Effects 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 208000030381 cutaneous melanoma Diseases 0.000 description 1
- 125000000151 cysteine group Chemical group N[C@@H](CS)C(=O)* 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000022811 deglycosylation Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- IBYVJTUUDGIOJT-UHFFFAOYSA-N dihydroperoxyborinic acid Chemical compound OOB(O)OO IBYVJTUUDGIOJT-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- SWSQBOPZIKWTGO-UHFFFAOYSA-N dimethylaminoamidine Natural products CN(C)C(N)=N SWSQBOPZIKWTGO-UHFFFAOYSA-N 0.000 description 1
- FSXRLASFHBWESK-UHFFFAOYSA-N dipeptide phenylalanyl-tyrosine Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FSXRLASFHBWESK-UHFFFAOYSA-N 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000006185 dispersion Substances 0.000 description 1
- 229930004069 diterpene Natural products 0.000 description 1
- 150000004141 diterpene derivatives Chemical class 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 238000004043 dyeing Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 210000000750 endocrine system Anatomy 0.000 description 1
- 201000003914 endometrial carcinoma Diseases 0.000 description 1
- 230000003511 endothelial effect Effects 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 201000004101 esophageal cancer Diseases 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000002270 exclusion chromatography Methods 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 230000001747 exhibiting effect Effects 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 239000012467 final product Substances 0.000 description 1
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 238000003500 gene array Methods 0.000 description 1
- 238000012637 gene transfection Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 208000005017 glioblastoma Diseases 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- 108010075431 glycyl-alanyl-phenylalanine Proteins 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 229960004198 guanidine Drugs 0.000 description 1
- 229960000789 guanidine hydrochloride Drugs 0.000 description 1
- PJJJBBJSCAKJQF-UHFFFAOYSA-N guanidinium chloride Chemical compound [Cl-].NC(N)=[NH2+] PJJJBBJSCAKJQF-UHFFFAOYSA-N 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 1
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 102000046949 human MSC Human genes 0.000 description 1
- 102000057305 human giant cell Human genes 0.000 description 1
- 108700016261 human giant cell Proteins 0.000 description 1
- 235000003642 hunger Nutrition 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000005847 immunogenicity Effects 0.000 description 1
- 230000001506 immunosuppresive effect Effects 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000000099 in vitro assay Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- ZPNFWUPYTFPOJU-LPYSRVMUSA-N iniprol Chemical compound C([C@H]1C(=O)NCC(=O)NCC(=O)N[C@H]2CSSC[C@H]3C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@H](C(N[C@H](C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC=4C=CC=CC=4)C(=O)N[C@@H](CC=4C=CC(O)=CC=4)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC=4C=CC=CC=4)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](CCCNC(N)=N)NC2=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CSSC[C@H](NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H]2N(CCC2)C(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N2[C@@H](CCC2)C(=O)N2[C@@H](CCC2)C(=O)N[C@@H](CC=2C=CC(O)=CC=2)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N2[C@@H](CCC2)C(=O)N3)C(=O)NCC(=O)NCC(=O)N[C@@H](C)C(O)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(=O)N[C@@H](CC=2C=CC=CC=2)C(=O)N[C@H](C(=O)N1)C(C)C)[C@@H](C)O)[C@@H](C)CC)=O)[C@@H](C)CC)C1=CC=C(O)C=C1 ZPNFWUPYTFPOJU-LPYSRVMUSA-N 0.000 description 1
- 230000000977 initiatory effect Effects 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000007912 intraperitoneal administration Methods 0.000 description 1
- 238000007913 intrathecal administration Methods 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 238000004255 ion exchange chromatography Methods 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 201000010982 kidney cancer Diseases 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 238000000670 ligand binding assay Methods 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 208000003747 lymphoid leukemia Diseases 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- -1 mannitol or sorbitol Chemical compound 0.000 description 1
- 201000001441 melanoma Diseases 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 238000001823 molecular biology technique Methods 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- AEMBWNDIEFEPTH-UHFFFAOYSA-N n-tert-butyl-n-ethylnitrous amide Chemical compound CCN(N=O)C(C)(C)C AEMBWNDIEFEPTH-UHFFFAOYSA-N 0.000 description 1
- 210000000822 natural killer cell Anatomy 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229960004927 neomycin Drugs 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 201000002575 ocular melanoma Diseases 0.000 description 1
- 238000006384 oligomerization reaction Methods 0.000 description 1
- 238000011275 oncology therapy Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 230000020477 pH reduction Effects 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 201000003913 parathyroid carcinoma Diseases 0.000 description 1
- 208000017954 parathyroid gland carcinoma Diseases 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 125000005496 phosphonium group Chemical group 0.000 description 1
- 208000021310 pituitary gland adenoma Diseases 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 102000021127 protein binding proteins Human genes 0.000 description 1
- 108091011138 protein binding proteins Proteins 0.000 description 1
- 230000007026 protein scission Effects 0.000 description 1
- 210000001938 protoplast Anatomy 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000006798 recombination Effects 0.000 description 1
- 238000005215 recombination Methods 0.000 description 1
- 206010038038 rectal cancer Diseases 0.000 description 1
- 201000001275 rectum cancer Diseases 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000012723 sample buffer Substances 0.000 description 1
- 239000004576 sand Substances 0.000 description 1
- 238000003118 sandwich ELISA Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 108010026333 seryl-proline Proteins 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 201000000849 skin cancer Diseases 0.000 description 1
- 201000003708 skin melanoma Diseases 0.000 description 1
- 201000002314 small intestine cancer Diseases 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 239000000176 sodium gluconate Substances 0.000 description 1
- 235000012207 sodium gluconate Nutrition 0.000 description 1
- 229940005574 sodium gluconate Drugs 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 210000000278 spinal cord Anatomy 0.000 description 1
- 206010041823 squamous cell carcinoma Diseases 0.000 description 1
- 230000037351 starvation Effects 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 125000000472 sulfonyl group Chemical group *S(*)(=O)=O 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 230000004797 therapeutic response Effects 0.000 description 1
- 150000003573 thiols Chemical class 0.000 description 1
- 108010071097 threonyl-lysyl-proline Proteins 0.000 description 1
- 201000002510 thyroid cancer Diseases 0.000 description 1
- 238000004448 titration Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 238000003151 transfection method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010020532 tyrosyl-proline Proteins 0.000 description 1
- 210000003606 umbilical vein Anatomy 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 201000005112 urinary bladder cancer Diseases 0.000 description 1
- 206010046766 uterine cancer Diseases 0.000 description 1
- 206010046885 vaginal cancer Diseases 0.000 description 1
- 208000013139 vaginal neoplasm Diseases 0.000 description 1
- 239000004474 valine Substances 0.000 description 1
- 108010072644 valyl-alanyl-prolyl-glycine Proteins 0.000 description 1
- 108010052774 valyl-lysyl-glycyl-phenylalanyl-tyrosine Proteins 0.000 description 1
- 108010009962 valyltyrosine Proteins 0.000 description 1
- 239000003981 vehicle Substances 0.000 description 1
- 201000005102 vulva cancer Diseases 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 108010027345 wheylin-1 peptide Proteins 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/46—Hybrid immunoglobulins
- C07K16/468—Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/02—Antineoplastic agents specific for leukemia
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/22—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against growth factors ; against growth regulators
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
- C07K2317/35—Valency
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/55—Fab or Fab'
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
- C07K2317/56—Immunoglobulins specific features characterized by immunoglobulin fragments variable (Fv) region, i.e. VH and/or VL
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/64—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising a combination of variable region and constant region components
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/60—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments
- C07K2317/66—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising a swap of domains, e.g. CH3-CH2, VH-CL or VL-CH1
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
- C07K2317/76—Antagonist effect on antigen, e.g. neutralization or inhibition of binding
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Immunology (AREA)
- Life Sciences & Earth Sciences (AREA)
- Medicinal Chemistry (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Animal Behavior & Ethology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Pharmacology & Pharmacy (AREA)
- General Engineering & Computer Science (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Hematology (AREA)
- Oncology (AREA)
- Plant Pathology (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Description
201111394 六、發明說明: 【發明所屬之技術領域】 本發明係關於一種雙特異性四價之抗原結合蛋白;其製 備方法、含有該等抗體之醫藥組合物、及其用途。 【先前技術】 * 於相關技術中已知諸如雙特異性或多特異性抗體之經工 程處理之蛋白質能夠結合兩種或兩種以上抗原。可利用細 胞融合術、化學結合術、或重組DNA技術產生該等多特異 性結合蛋白。 最近幾年,已發展出許多種形式之重組多特異性抗體, 例如藉由使例如IgG抗體形式與單鏈結構域融合產生四價 雙特異性抗體(參見例如Coloma,M.J·,等人,Nature Biotech. 15(1997)159-163 ; WO 2001/077342 ;及 Morrison, S.L.,Nature Biotech. 25(2007)1233-1234)。 亦已發展出許多種其他新穎形式,其不再保留抗體核心 結構(IgA、IgD、IgE、IgG或IgM),諸如雙功能抗體、三 功能抗體或四功能抗體、微型抗體、若干種單鏈形式 (scFv、Bis-scFv),且其能夠結合兩種或兩種以上抗原 . (Holliger,P·,等人,Nature Biotech 23(2005)1 126-1 136 ;
Fischer, N·,及 L6ger, 0., Pathobiology 74(2007)3-14 ; Shen, J.,等人,J. Immunol. Methods 318(2007)65-74 ; Wu,C·,等人,Nature Biotech· 25(2007)1290-1297)。 所有該等形式均使用連接子,以使抗體核心(IgA、 IgD、IgE、IgG或IgM)與其他結合蛋白(例如scFv)融合,或 I48423.doc 201111394 使例如:兩個Fab片段或scFv融合(Fischer,N•,及
Path〇bi〇l〇gy 74(20〇7)3-14)。雖然連接子顯然有利於 工程處理雙特異性抗體,但是其亦可於治療中引發問題。 事貫上,該等外來肽可能引發針對連接子本身、或蛋白質 與連接子之間的連接部份的免疫反應。更進一步,該等肽 之可撓性使其更容易發生蛋白質裂解,且可能導致抗體穩 疋性差、凝集及免疫原性增加。此外,相關技術者可能希 望藉由保持與天然存在抗體高度相似,保留透過Fc部份所 介導之效應子功能,諸如例如依賴補體之細胞毒性作用 (CDC)或依賴抗體之細胞毒性作用(adcc)。 因此,理想地,相關技術者意欲發展出一種雙特異性抗 體,其與天然存在之抗體(如IgA、igD、IgE、IgG或IgM) 的一般結構極類似,且與人類序列的不同處最少。 於一方法中,已利用基於使兩種不同融合瘤細胞株(表 現出具所需之雙特異性抗體之特異性的鼠科單株抗體)進 行體細胞融合的四源雜交瘤技術(參見MUstein,c.,及
Cuello,A.C·,Nature 305(1983)537-540),製得與天然抗體 極相似之雙特異性抗體。由於在所得之雜交-融合瘤(或四 源雜交瘤)細胞株中’兩種不同之抗體重鏈及輕鏈隨機配 對’會產生至多10種不同抗體種類,其中僅有一種為所需 之功能性雙特異性抗體。由於存在錯誤配對之副產物,而 且生產率顯著降低,意味著需要複雜之純化製程(參見例 如 Morrison,S.L·,Nature Biotech. 25(2007)1233-1234)。若 使用重組表現技術’通常仍會產生錯誤配對之副產物的相 I48423.doc 201111394 同問題。 > 一種解決存在錯誤配對之副產物問題的方法稱為「結-入-穴」(knobs-into-holes),其意欲向CH3結構域引入突變 而改變接觸介面,驅使兩種不同之抗體重鏈配成對。於一 條鏈中,由具短側鏈之胺基酸替代大型胺基酸,產生 「穴」。反之’向另一 CH3結構域引入具大型側鏈之胺基 酸’產生「結」。藉由共同表現該等兩種重鏈(及兩條必需 適於兩條重鏈之相同輕鏈),高產量形成雜二聚體(「結_ 穴」(knob-hole)),而並非同二聚體(「穴-穴」(h〇ie_hole) 或「結-結」(knob-knob))(Ridgway,J.B·,等人,Protein
Eng. 9(1996)617-621 ;及 WO 96/027011)。可利用噬菌體 呈現術重塑兩個CH3結構域之相互作用表面,並引入雙硫 橋來穩疋雜二聚體,進一步增加雜二聚體之百分比 (Merchant,A.M,等人,Nature Biotech 16(1998)677-681 ;
Atwell,S.,等人,j· Mol Bi〇1 27〇(1997)26_35)。於例如 EP 1 870 459 A1中,闡述一種進行結_入_穴技術之新穎方 法。雖然該方法非常吸引人,但是目前仍無法獲得關於臨 床上進展的數據。該方法之一重要限制點為兩種母本抗體 的輕鏈必須相’以防止錯誤配對及形成無活性分子。因 此,該技術不適於自對抗第—及第二抗原的兩種抗體輕易 發展出對抗兩種抗原之重組雙特異性抗體,因為該等抗體 之重鏈及/或相同之輕鏈必須經過最適化。當例如藉由使 另外之Fab片段(分別與可結合第一或第二抗原之μ片段 相同)與雜一聚體之各重鏈c端融合,改表現出四價雙特異 148423.doc 201111394 性抗體來替代雙價雙特異性抗體時,仍然存在相同之錯誤 配對問題(參見圖2)。 WO 2006/093794係關於一種雜二聚體蛋白質結合組合 物。WO 99/37791闞述一種多標的抗體衍生物。M〇rris〇n, S.L.,等人,the J. Immunolog,160(1998)2802-2808提及交 換可變區結構域對IgG之功能性質之影響。 【發明内容】 本發明包括一種雙特異性四價之抗原結合蛋白質,其包 括: a) 第一抗體之經修飾重鏈, 其可與第一抗原特異性結合, 且違重鍵之C端另與該第一抗體之vh-CH 1結構域的n 端融合; b) 如a)之該第一抗體之兩條輕鏈; c) 第·一抗體之經修飾重鏈, 其可與第二抗原特異性結合, 其中CH1結構域被該第二抗體之cl結構域置換, 且該重鏈之C端另與該第二抗體之Vh-CL結構域的N 端融合;及 d) 如c)之β亥第一抗體之兩條經修飾輕键, 其中CL結構域被該第二抗體之CH1結構域置換。 本發明之另一實施例為一種製備根據本發明之抗原結合 蛋白的方法, 包括如下步驟: 148423.doc • 6 - 201111394 a) 以包括可編碼根據本發明之雙 枋納八2 付旲11抗原結合蛋白的 核i刀子之載體轉形宿主細胞; b) 於允許合成該抗體分子之 、ώ ¥ 旰卜培養宿主細胞;及 c) 自β亥培養物收集該抗體分子。 本發明之另-實施例為一種宿主細胞,其包括 • 一種包括可編碼根據本發明 :柷原結合蛋白之核酸分 子的載體。 本發明之另一實施例為一種醫藥 ^ H t 裡请樂組合物’其包括根據本 發明之抗原結合蛋白及至少一種醫 裡请樂上可接受賦形劑。 本發明之另一實施例為一種治療需要治療之患者的方 法,其特徵為向患者投與治療有效* 結合蛋自。 有效量之輯本發明之抗原 根據本發明’可藉由如下方法改善所需之雙特異性四價 之杬原結合蛋白對不需要之副產物的比例:於如C)之經修 飾重鏈中,且亦於如d)對應之兩條經料之輕鏈中,以CL 結構域置換cm。依此方式,可減少可特異性結合第一抗 原之抗體輕鏈⑻與可特異性結合第二抗原之抗體的經修飾 抗體重鍵⑷的錯誤VH-CH1結構域之間的不期望之錯誤配 對。且同理可減少如d)之經修飾輕鏈與如a)之重鍵的錯誤 配對。(參見圖2,其無該等修飾且錯誤配對;及圖3,其 具有該f CH1-CL置換(或交換)且沒有錯誤配對)。 【實施方式】 括 本發明包括-種雙特異性四價之抗原結合蛋白,其包 148423.doc 201111394 a) 第一抗體之經修飾重鏈, 其可與第一抗原特異性結合, 且该重鏈之c端另經肽連接子與該第一抗體中之VH_ CH1結構域的N端融合; b) 兩條如a)之該第一抗體之輕鏈; c) 第一抗體之經修飾重鏈, 其可與第二抗原特異性結合, 其中CH1結構域被該第二抗體之CL結構域置換, 且5亥重鏈之c端另經肽連接子與該第二抗體中之VH_ CL結構域的n端融合;及 d) 如c)之該第一抗體之兩條經修飾之輕鍵, 其中構域被該第二抗體之CH1結構域置換。 根據本發明’可藉由如下方法改善所需之雙特異性四價 之抗原、.、。&蛋白對不需要之副產物(由於抗體輕鏈與錯誤 的杬體重鏈之間的錯誤配對)之比例:於如匀之經修飾重鏈 中及於如d)之對應兩條經修飾之輕鏈中,以結構域置換 CH1。此時之錯誤配對意指締合:〇可特異性結合第一抗 原b)之抗體的輕鏈與可特異性結合第二抗原之抗體的經修 飾之重鏈;或ii)可特異性結合第二抗原b)之抗體的輕鏈與 可特異性結合第一抗原之抗體的經修飾之重鏈(參見圖2), 其產生不需要之無活性或無完整功能性之副產物。 於本發明之另一態樣中,所需之雙特異性四價抗體對不 需要之副產物之經改善之比例可藉由如下方法進一步改 °修飾可特異性結合第一及第二抗原之該等抗體之CH3 I48423.doc 201111394 結構域。 因此’於本發明之一項較佳實施例中,可藉由「結-入· 穴」(knob-into-holes)技術改變根據本發明之該雙特異性 四價之抗原結合蛋白之CH3結構域(於如&及c之經修飾重鏈 中),該技術詳細闡述於一些實例中,例如w〇 96/027011 ; Ridgway,J.B·,等人,Protein Eng 9(1996) 617-621 ;及 Merchant,A.M·,等人,Nat Biotechnol 16 (1998)677-681。於該方法中,改變兩個cH3結構域之相互 作用表面’以使含有該等兩種CH3結構域之兩條重鏈增加 雜一聚化。兩條重鏈中之兩個CH3結構域中分別可為 「結」(knob),而另一則可為「穴」(h〇ie)。引入雙硫橋, 進一步穩定雜二聚體(Merchant,A.M.,等人,Nature
Biotech 16(1998)677-681 ; Atwell, S.,等人,J. Mol· Biol. 270(1997)26-35)並增加產量。 因此’於本發明之一態樣中’該雙特異性四價之抗原結 合蛋白之其他特徵為: a)之抗體之經修飾重鏈的CH3結構域與c)之抗體之經修 飾重鏈的CH3結構域各在一個包含該等抗體CH3結構域之 間原始介面之介面會合(meet); -其中該介面經改變以促進形成該雙特異性之四價抗原 結合蛋白,其中該改變之特徵為: i) 一條重鏈之CH3結構域改變, 使知在該雙特異性四價之抗原結合蛋白中與另一條重鏈 之CH3結構域之原始介面會合之一條重鏈之CH3結構域之 148423.doc -9- 201111394 原始介面内, 一個胺基酸殘基被一個具有較大側鏈體積之胺基酸殘基 置換,由此在一條重鏈之CH3結構域之介面内產生一個突 起,其可置入另一條重鏈之CH3結構域之介面内之一個孔 穴中 及 ii)另一條重鏈之CH3結構域改變, 使得在該雙特異性四價之抗原結合蛋白中與該第一 CH3 結構域之原始介面會合之該第二CH3結構域之原始介面 内, 一個胺基酸殘基被一個具有較小側鏈體積之胺基酸殘基 置換’由此於該第二CH3結構域之介面内產生一個孔穴, 其中可置入該第一 CH3結構域之介面内之一個突起。 具有較大側鏈體積之該胺基酸殘基較佳係自由以下組成 之群中選出:精胺酸(R)、苯丙胺酸、酪胺酸、色胺 酸(W) 〇 具有較小側鏈體積之該胺基酸殘基較佳係自由以下組成 之群中選出:丙胺酸(A)、絲胺酸(S)、蘇胺酸(T)、纈胺酸 (V) 〇 於本發明之一態樣中’進一步改變兩個CH3結構域,其 係引入半胱胺酸(C)作為各CH3結構域中之對應位點的胺基 酸,使得於兩個CH3結構域之間形成雙硫橋。 於一項較佳實施例中’該雙特異性四價之抗原結合蛋白 包括在「結鏈」(knobs chain)之CH3結構域中之T366W突 148423.doc -10- 201111394 變、及在「穴鏈」(hole chain)之CH3結構域中之T366S、 L368A、Y407V突變。於CH3結構域之間亦可使用其他鏈 間雙疏橋(Merchant, Α·Μ.,等人,Nature Biotech. 16 (1998)677-681),例如在「結鏈」或「穴鏈」之CH3結構 域内引入Y349C突變、及於另一鏈之CH3結構域内引入 E3 5 6C突變或S354C突變。因此,於另一項較佳實施例 中,該雙特異性四價之抗原結合蛋白在兩個CH3結構域中 一個結構域包括S354C、T366W突變;及在兩個CH3結構 域中另一個結構域包括Y349C、T366S、L368A、Y407V突 變;或者該雙特異性四價之抗原結合蛋白在兩個CH3結構 域中一個結構域包括E356C、T366W突變;及在兩個CH3 結構域中另一個結構域包括Y349C、T366S、L368A、 Y407V突變;或者該雙特異性四價之抗原結合蛋白在兩個 CH3結構域中一個結構域包括Y349C、T366W突變;及在 兩個CH3結構域中另一個結構域包括E356C、T366S、 L368A、Y407V突變;或者該雙特異性四價之抗原結合蛋 白在兩個CH3結構域中一個結構域包括Y349C、T366W突 變;及在兩個CH3結構域中另一個結構域包括S354C、 T366S、L368A、Y407V突變;(其中一個CH3結構域再包 括Y349C突變,另一個CH3結構域中再包括E356C或S354C 突變,形成鏈間雙硫橋)(根據Kabat EU目錄進行所有編 號)。但亦可改用或另外使用如EP 1 870 459 A1所述之其 他結-入-穴技術。該三特異性或四特異性抗體之較佳實例 包括於「結鏈」之CH3結構域中之R409D、K370E突變; 148423.doc 201111394 及於「穴鏈」之CH3結構域中之D399K、E357K突變(根據 Kabat EU目錄進行所有編號)。 於另一較佳實施例中,該三特異性或四特異性抗體包括 於「結鏈」之CH3結構域中之T366W突變、及於「穴鏈」 之CH3結構域中之T366S、L368A、Y407V突變,且另外包 括於「結鏈」之CH3結構域中之R409D、K370E突變;及 於「穴鏈」之CH3結構域中之D399K、E357K突變。 於另一較佳實施例中,該三特異性或四特異性抗體在兩 個CH3結構域中一個結構域包括Y349C、T366W突變;及 在兩個CH3結構域中另一個結構域包括S354C、T366S、 L3 68A、Y407V突變;或者該三特異性或四特異性抗體在 兩個CH3結構域中一個結構域包括Y349C、T366W突變; 及在兩個CH3結構域中另一個結構域包括S354C、T366S、 L368A、Y407V突變,且另外包括於「結鏈」之CH3結構 域中包括R409D、K3 70E突變;及於「穴鏈」之CH3結構 域中包括D3 99K、E357K突變。 如文中所用,術語「抗體」係指由兩條抗體重鏈及兩條 抗體輕鏈組成之全長抗體(參見圖1)。全長抗體之重鏈為自 N端至C端方向,由如下部份所組成之多肽:抗體重鏈可 變結構域(VH)、抗體恆定重鏈結構域1(CH1)、抗體鉸鏈區 (HR)、抗體重鏈恆定結構域2(CH2)、及抗體重鏈恆定結構 域 3(CH3),縮寫為 VH-CH1-HR-CH2-CH3 ;且若抗體為 IgE 子組,則視需要包括抗體重鏈恆定結構域4(CH4)。全長抗 體之重鏈較佳為自N端至C端方向由如下部份所組成之多 148423.doc •12· 201111394 肽:vh、CH1、HR、CH2及CH3。全長抗體之輕鏈為自N 端至C端方向由如下部份所組成之多肽:抗體輕鏈可變結 構域(VL)、抗體輕鏈恆定結構域(CL),縮寫為vL_CL。抗 體輕鏈恆定結構域(CL)可為K(kappa)4Mlambda)型。抗體 鏈係經CL結構域與CH1結構域之間(亦即輕鏈與重鏈之 間)' 及全長抗體重鏈之鉸鏈區之間的多肽之間雙硫橋鍵 結在一起。典型全長抗體實例為天然抗體,諸如igG(例如 IgGl及IgG2)、IgM、IgA、IgD、及lgE。根據本發明之抗 體可源自單一物種(例如人類),或者其可為嵌合或人類化 抗體。根據本發明之全長抗體包括兩個抗原結合位點,其 分別由一對VH及VL形成,其二者皆特異性結合相同(第 一)抗原。該全長抗體之重鏈或輕鏈的c端係指於該重鏈或 輕鏈之C端之最後一個胺基酸。該抗體包括兩個相同的Fab 片段,其係由重鏈之VH及CH1結構域及輕鏈之及CL結 構域組成(參見圖1)。 於根據本發明之抗原結合蛋白中,可特異性結合第二抗 原之第二抗體之CH1&CL結構域相互置換產生如d)之經 修飾輕鏈及如c)之經修飾抗體重鏈,其中該重鏈之c端再 經由肽連接子與該抗體(可特異性結合第二抗原)之vh_cl 結構域之N端融合。 該抗體(可特異性結合第一抗原)之「VH_CH1結構域」 係指依N端至C端方向之該抗體之VH& cm結構域。且該 抗體(可特異性結合第二抗原)之「乂11_(:(:[結構域」係指 依N端至C端方向之該抗體之VH&CH1結構域。 I48423.doc •13- 201111394 如本發明所用,術語「肽連接子」係指具有較佳為合成 性起源之胺基酸序列之肽。該等根據本發明之肽連接子係 用於使抗原結合肽與全長及/或經修飾之全長抗體鏈的c端 或N端融合,以形成根據本發明之雙特異性抗原結合蛋 白如c)中之該肽連接子較佳為具有長度為至少$個胺基 酸,較佳長度為5至100個,更佳1〇至5〇個胺基酸之胺基酸 序列之肽。於一項實施例中,該肽連接子為(GxS)n或 (GxS)nGm ’其中G=甘胺酸,s=絲胺酸,且(χ=3,n=3、 4、5 或 6,及 m=〇、】、2 或 3)或(χ=4,n=2、3、4 或 5 及 m 1、2或3) ’較佳χ=4及n=2或3,更佳x=4,n=2。於 一項實施例中,該肽連接子為(g4S)2。 如文中所用,術語「結合位點」或「抗原結合位點」係 和於根據本發明之抗原結合蛋白中之實際上與配位體(例 如抗原或其抗原片段)相結合之區域,且其係衍生自抗體 分子或其片段(例如Fab片段)。根據本發明之抗原結合位點 匕括"T與所需抗原特異性結合之抗體或抗體片段中的抗體 重鏈可變結構域(VH)及抗體輕鏈可變結構域(VL)。 可與所需抗原特異性結合之抗原結合位點(亦即成對之 VH/VL)可讨生自a)針對該抗原之已知抗體’或b)利用特定 言之抗原蛋白或核酸或其片段進行重新合成免疫接種方 法’或藉由噬菌體呈現術所獲得之新穎抗體或抗體片段。 本發明之抗原結合蛋白之抗原結合位點含有六個互補決 定區(CDR),其使結合位點對抗原產生不同程度之親和 力。有三個重鏈可變結構域CDR(CDRH1 ' CDRH2及 148423.doc 201111394 CDRH3)及三個輕鏈可變結構域CDR(CDRL1、CDRL2& CDRL3)。CDR及框架區域(FR)之範圍的確定方法為藉由 與已編譯之胺基酸序列資料庫比較,其中已根據序列之間 的可變性定義該等區域。 抗體特異性係指抗體或抗原結合蛋白對抗原中之特定抗 原決定基的選擇識別性。例如天然抗體具單特異性。雙特 異性抗體為具有兩種不同抗原結合特異性之抗體。當抗體 具有一種以上特異性時,所識別之抗原決定基可與單一抗 原或與一種以上抗原結合。 如文中所用,術語「單特異性」抗體或抗原結合蛋白係 扣具有-個或多個可結合相同抗原之相同抗原決定基的抗 體或抗原結合蛋白。 如本申請案所用,術言吾「價態」係指於抗體分子中所存 在之結合位點的明確數量。例如天然抗體或根據本發明之 全長抗體具有兩個結合位點,且為兩價。術語「四價」係 指於抗原結合蛋白中存在四個結合位點。如文中所用術 。。雙特異性四仏」係指根據本發明之抗原結合蛋白,其 具有四個抗原結合位點,且其中兩個結合第一抗原,且另 外兩個„第—抗原(或抗原之另—抗原決定基)。本發明 之抗原結合蛋白具有四個結合位點,且為四價。 本發明之全長抗體肖杯厘认 括屬於一種或多種免疫球蛋白類之 免疫球蛋白以區。免疫球蛋白類包括邮、响、心、 g及1gE同型’且右為IgG及IgA,則存在亞型。於一項 較佳實施例中,本發明之全長抗體具有邮型抗體之值定 148423.doc 201111394 結構域結構。 如文中所用,術語「單株抗體」或「單株抗體組合物」 係指具單一胺基酸組成之抗體製劑或抗體或抗原結合蛋白 分子。 術语「嵌合抗體」係指包括源自一種來源或物種之可變 區(亦即結合區)、及源自不同來源或物種之恒定區之至少 一部份的抗體,其通常係藉由重組DNA技術製得。以包括 齧齒類動物之可變區及人類恆定區的嵌合抗體為較佳。本 發明所涵蓋之其他較佳形式之「嵌合抗體」為彼等其中恆 疋區經修飾或改變而不同於原始抗體之恆定區以產生根 據本發明之性質,尤其指Clq結合性及/或卜受體(FcR)結 合性。該等嵌合抗體亦稱為「種類轉換抗體」。嵌合抗體 為由免疫球蛋白基因所表現之產物,該等免疫球蛋白基因 包括編碼免疫球蛋白可變區之DNA片段及編碼免疫球蛋白 恆定區之DNA片段。製備嵌合抗體之方法涉及習知之重組 DNA及基因轉染技術,且其係於相關技術中已知。參見例 如 Morrison, S丄.,等人,Proc. Natl Acad Sci USA 81 (1984)6851-6855 ; US 5,202,238及 US 5,204,244。 術語「人類化抗體」係指其中框架區或「互補決定區」 (CDR)經修飾,以包括與母本免疫球蛋白具有不同特異性 之免疫球蛋白的CDR。於一項較佳實施例中,將鼠CDR^g 植至人類抗體之框架區中,製得「人類化抗體」。參見例 如 Riechmann,L.,等人,Nature 332(1988)323-327 ;及 Neuberger,M.S·,等人 ’ Nature 314(1985)268-270。特別 J48423.doc • 16 · 201111394 佳之CDR對應於彼等代表可識別針對嵌合抗體之上述抗原 之序列。本發明所涵蓋之其他形式之「人類化抗體」為彼 等其中恆定區已經進一步修飾或改變而不同於原始抗體之 恆定區’以產生根據本發明之性質,尤指C1q結合性及/或 Fc受體(FcR)結合性。 如文中所用’術語「人類抗體」意欲包括具有衍生自人 類生殖系(germ line)免疫球蛋白序列之可變區及恆定區之 抗體。人類抗體係相關技藝已熟知(van Dijk,Μ. A.,及van de Winkel,J.G·,Curr. Opin. Chem_ Biol. 5(2001)368-374)。 人類抗體亦可於轉殖基因動物(例如小鼠)中製得,該等轉 殖基因動物能夠在免疫時,在不產生内源性免疫球蛋白 下,產生全譜(repertoire)或選擇之人類抗體。人類生殖系 免疫球蛋白基因陣列轉移入該生殖系突變小鼠中會導致在 才;υ原激發時產生人類抗體(參見例如Jakobovits,A.,等人, Proc. Natl. Acad. Sci. USA 90(1993)25 5 1-25 55 ;
Jakobovits, A.,等人,Nature 362(1993)255-258 ;
Brueggemann,M.,等人,Year Immunol. 7(1993)33-40)。 人類抗體亦可於嗟菌體呈現庫中製得(Hoogenboom,H.R., 及 Winter,G.J.,Mol. Biol. 227(1992)381-388 ; Marks,J.D., 等人 ’ J. Mol. Biol. 222(1991)581-597)。Cole,S.P.C.,等 人及Boerner等人之技術亦可用於製備人類單株抗體(c〇le, S.P.C.,等人,Monoclonal Antibodies and Cancer Therapy, Alan R. Liss,第 77 頁(1985);及 Boerner, P.,等人,_1· Immunol. 147(1991)86-95)。如已針對根據本發明之嵌合及 148423.doc 201111394 人類化抗體所述,如文中所用,術語「人類抗體」亦包含 例如藉由「種類轉換」(class switching),亦即使Fc部份改 變或突變(例如自IgGl形成IgG4及/或lgGl/IgG4突變),在 怪定區中進行修飾,以產生根據本發明之性質,尤其Clq 結合及/或FcR結合之抗體。 如文中所用’術語「重組人類抗體」意欲包括藉由重組 方法製備、表現、產生或分離之所有人類抗體,諸如自宿 主細胞(諸如NS0或CHO細胞)或自轉殖人類免疫球蛋白基 因之動物(例如小鼠)分離之抗體.,或利用重組表現載體轉 染宿主細胞而表現之抗體。該等重組人類抗體具有重排形 式之可變區及悝定區。根據本發明 < 重組人類抗體已經活 體内體細胞超突變。因此,重組抗體之VH及VL區之胺基 酸序列雖然係衍生自且相關於人類生殖系VH及VL序列, 但可能天然不存在於活體内人類抗體生殖系譜中。 如文中所用’「可變結構域」(輕鏈之可變結構域(VL)、 重鏈之可變結構域(VH))係指直接參與抗體與抗原結合之 每一對輕鏈及重鏈中之結構域。可變人類輕鏈及重鏈結構 域具有相同之一般結構,且各結構域包括四個框架區 (FR) ’其序列高度保守’且其係由三個「高可變區」(或 互補決定區、CDR)連接。框架區採取β_折叠片構形,且 CDR可形成連接β-折叠片結構體之環。各鏈中CDR之三維 結構係由框架區保持,且與來自另一條鏈2CDR—起形成 抗原結合位點。抗體重鏈及輕鏈CDR3區在根據本發明之 抗體的結合特異性/親和力上扮演特別重要之角色,且因 148423.doc • 18- 201111394 此提供本發明之另一標的。 如文中所用’術語「高可變區」或「抗體之抗原結合部 份」係指抗體中負責與抗原結合之胺基酸殘基。高可變區 包括「互補決定區」或r CDR」之胺基酸殘基。「框架 區」或「FR」區為彼等除了文中所定義之高可變區殘基以 外的可變結構域區域。因此,抗體之輕鏈及重鏈包括自N 端至C端之如下結構域:FR1、CDR1、FR2、CDR2、 FR3、CDR3、及FR4。藉由該等框架胺基酸分離各鏈上之 CDR。特別地,重鏈之CDR3為對抗原結合性影響最大之 Q 域 根據 Kabat,專人,Sequences of Proteins of
Immunological Interest,第 5 版 ’ Public Health Service, National Institutes of Health,Bethesda,MD(1991)之標準定 義決定CDR及FR區。 如文中所用,術語「結合」、「特異性結合」、或「其特 異性結合」係指於活體外分析中,抗體/抗原結合蛋白與 抗原之抗原決定基結合,較佳為在電漿共振分析(BIAc〇re, GE-Healthcare Uppsala,Sweden)中,與經純化之野生型抗 原結合。該結合親和力係由術語ka(抗體/抗原複合物中抗 體(或抗體或抗原結合蛋白)之締合速率常數)、kD(解離常 數)、及KD(kD/ka)定義。結合或特異性結合意指結合親和 力(kd)為1〇·8 m〇w或以下,較佳1〇-9]^至1〇-丨3 m〇m。因 此,根據本發明之雙特異性四價之抗原結合蛋白可與各特 異性抗原特異性結合,且結合親和力(尺〇)為1〇·8爪“/丨或以 下’較佳 ΙΟ·9 Μ至 10-n mol/1。 I48423.doc -19- 201111394 可藉由 BIAcore分析法(GE_Healthcare Uppsaia,瑞典)研 究抗體與FcYRIII之結合。該結合親和力係由術語匕(抗體/ 抗原複合物中抗體之締合速率常數)、kD(解離常數)、及 KD(kD/ka)定義。 術語「抗原決定基」包括能夠與抗體特異性結合之任一 多肽決定基。於某些實施例中,抗原決定基包括具化學活 性之表面分子基團,諸如胺基酸、糖侧鏈、磷醯基、或磺 醯基,且於某些實施例中,其可具有特異性三維結構特 徵,或特異性電荷特徵。抗原決定基為抗原中與抗體結合 之區域。 於某些實施例中,當抗體在蛋白質及/或大分子物之複 雜混合物中優先識別其標靶抗原時,稱該抗體係與抗原特 異性結合。 於另一實施例中,根據本發明之雙特異性抗原結合蛋白 之特徵為:該全長抗體屬於人類IgG1亞類,或屬於具有突 變L234A及L235A之人類igG1亞類。 於另一實施例中,根據本發明之雙特異性抗原結合蛋白 之特徵為:該全長抗體屬於人類IgG2亞類。 於另一實施例中,根據本發明之雙特異性抗原結合蛋白 之特徵為:該全長抗體屬於人類IgG3亞類。 於另一實施例中,根據本發明之雙特異性抗原結合蛋白 之特徵為:該全長抗體屬於人類IgG4亞類、或具有另一突 變S228P之人類IgG4亞類。 争乂佳地’根據本發明之雙特異性抗原結合蛋白之特徵 148423.d〇c -20- 201111394 為.該全長抗體屬於人類1凸麵 η . 八頰igui亞頒、具有另一突變S228p 之人類IgG4亞類。 現已發現’根據本發明之雙特異性抗原結合蛋白具有改 善之特徵’諸如生物學或藥理學活性、藥物動力學性質或 毒性。其可用於例如治療諸如癌症之疾病。 如本申請案所用,術語「恆定區」係指抗體中除了可變 區之外之全部結構域。恆定區不直接參與與抗原之結合, 但顯示多種效應子功能。取決於重鏈恆定區之胺基酸序 列,將抗體分成如下幾組:IgA、IgD、IgE、砂及_, 且其中一些可進一步分成子組,諸如IgGl、IgG2、IgG3、 及IgG4、IgAl及IgA2。對應於不同抗體組之重鏈恆定區分 別稱為α、δ、ε、γ、及μ。存在於所有五種抗體組之輕鏈 值定區(CL)稱為 K(kappa)及 X(Iambda)。 如本申請案所用,術語「源自人類來源之恆定區」係指 屬於IgGl、IgG2、IgG3、或IgG4子組之人類抗體之怪定重 鏈區域及/或恆定輕鏈κ或λ區域《該等恆定區係當前技術 中已熟知’且係例如由Kabat, Ε.Α.闡述(參見例如Johnson, G.及 Wu,T.T.,Nucleic Acids Res. 28(2000)214-218 ; Kabat, E.A.,等人,Proc· Natl. Acad. Sci. USA 72(1975)2785-2788)中。 雖然屬於IgG4亞類之抗體顯示下降之Fc受體(FcyRIIIa) 結合力,但屬於其他IgG亞類之抗體仍顯示強力結合力。 然而,如下殘基(若改變)亦可降低Fc受體結合力: Pro23 8、Asp265、Asp270、Asn297(喪失 Fc碳水化合物)、 148423.doc 21 201111394
Pro329、Leu234、Leu235、Gly236、Gly237、Ile253、 Ser254 、Lys288 、Thr307 、Gln311 、Asn434 、及 His435(Shields, R.L.,等人,J. Biol. Chem. 276(2001) 6591-6604 ; Lund, J·,等人,FASEB J. 9(1995) 1 15-1 19 ; Morgan, A.,等人,Immunology 86(1995)319-324 ; EP 0 307 434) ° 於一項實施例中,根據本發明之抗原結合蛋白具有比 IgGl抗體降低之FcR結合力,且與FcR結合力相關之其全 長母本抗體為在S228、L234、L235及/或D265處具有突 變、及/或含有PVA236突變之IgG4亞類或IgGl或IgG2亞 類。於一項實施例中,全長母本抗體中之突變為S228P、 L234A、L235A、L235E及 / 或 PVA236。於另一項實施例 中,全長母本抗體中之突變位於IgG4 S228P處及IgGl L234A及 L235A處。 抗體之恆定區直接參與ADCC(依賴抗體之由細胞介導之 細胞毒性作用)及CDC(依賴補體之細胞毒性作用)。補體活 化作用(CDC)係由補體因子Clq與大多數IgG抗體子組之恆 定區結合所引發。由位於所謂的結合位點處之所界定之蛋 白質-蛋白質相互作用引發Clq與抗體結合。該等恆定區結 合位點係相關技藝中已知,且闡述於例如如下文獻中: Lukas,T.J·,等人,J. Immunol. 127(1981)2555-2560 ; Brunhouse,R.及 Cebra,J.J·,Mol. Immunol. 16(1979)907-917 ; Burton, D.R·,等人,Nature 288(1980)338-344 ; Thomason, J.E.,等人,Mol. Immunol. 37(2000)995- 148423.doc • 11· 201111394 1004 ; Idusogie, E.E.,等人’】.11111111111〇1.164(2000)4178-4184 ; Hezareh, M.,等人,】.\^1*〇1.75(2001)12161_ 12168 ; Morgan, Α·,等人,Immunology 86(1995)319- 324 ;及EP 0 307 434。該等恆定區結合位點之特徵為例如 具胺基酸 L234、L235、D270、N297、E318、K320、 K322、P331、及P329(根據Kabat EU目錄編號)。 術語「依賴抗體之細胞毒性作用(ADCC)」係指在效應 子細胞存在下,由根據本發明之抗原結合蛋白裂解人類標 靶細胞。ADCC之較佳測定方法為:在效應子細胞(諸如新 鮮分離得之PBMC、或自白血球層純化所得之效應子細胞 (如單核細胞或天然殺手(NK)細胞)、或無限生長之NK細胞 株)存在下,以根據本發明之抗原結合蛋白處理可表現抗 原之細胞製劑。 術語「依賴補體之細胞毒性作用(CDC)」係指由補體因 子Clq與大多數igG抗體亞類之Fc部份之間的結合所引發之 過程。C 1 q與抗體之結合係由位於所謂之結合位點處所界 定之蛋白質-蛋白質相互作用所導致。該等Fc部份結合位 點係相關技術中已知(參見上文)。該等以部份結合位點之 特徵為例如具胺基酸L234、L235、D270、N297、E318、 K320、K322、P331、及P329(根據 Kabat £ϋ 目錄編號)。
IgGl、IgG2、及IgG3子組之抗體通常顯示包括與ciq及C3 結合之補體活化作用’而IgG4不會活化補體系統,且不與 Clq及/或C3結合。 可如 Umana,P.,等人,Nature Biotechnol. 17(1999)176- I48423.doc -23- 201111394 180 ;及US 6,6〇2,684中所述,藉由工程處理單株抗體之寡 糖組分,可強化單株抗體之由細胞介導之效應子功能。 IgGl類型抗體(最常使用之治療用抗體)為醣蛋白,其在各 CH2結構域之Asn297處均具有保守之與N鍵連之醣基化位 點。附接至Asn297之兩個複合之雙觸寡糖埋藏於CH2結構 域之中,形成與多肽主鏈的深度接觸,且其存在性係抗體 介導效應子功能(諸如依賴抗體之細胞毒性作用(ADCC))所 必須(Lifely,M., R.,等人,Glycobiology 5(1995)813-822 ; Jefferis,R.,等人,Immunol. Rev. 163(1998)59-76 ;
Wright, A.、及 Morrison,S·,L·,Trends Biotechnol. (1997)26-32)。Umana,P.,等人,Nature Biotechnol. 17(1999)176-180及WO 99/54342顯示,於中國倉鼠卵巢 (CHO)細胞中過度表現之β(1,4)-Ν-乙醯基葡糖胺基轉移酶 III(「GnTIII」)(一種催化形成等分寡糖之糖基轉移酶)顯 著增加抗體之活體外ADCC活性。改變Asn297碳水化合物 之組成或將其消除亦影響與FcyR及Clq之結合(Umana, P., 等人,Nature Biotechnol. 17(1999)176-180 ; Davies, J., 等人,Biotechnol· Bioeng. 74(2001)288-294 ; Mimura, Y., 等人,J. Biol. Chem. 276(2001)45539-45547 ; Radaev,S., 等人,J_ Biol. Chem. 276(2001)16478-16483 ; Shields,R·, L·,等人,J. Biol. Chem. 276(2001)6591-6604 ; Shields, R., L.,等人,J· Biol. Chem· 277(2002)26733-26740 ; Simmons, L_, C.,等人,J. Immunol. Methods 263(2002) 133-147)。 148423.doc •24- 201111394
於例如如下文獻中已報導增強單株抗體之由細胞介導之 效應子功能之方法 :WO 2005/018572 、 WO 2006/116260、WO 2006/114700、WO 2004/065540、WO 2005/01 1735 ' WO 2005/027966、WO 1997/028267、US 2006/0134709、US 2005/0054048 ' US 2005/0152894、WO 2003/035835 ' WO 2000/061739 ° 於本發明之一項較佳實施例中,雙特異性抗原結合蛋白 係經醣基化(若其包括IgGl、IgG2、IgG3或IgG4亞類,較 佳IgGl或IgG3亞類之Fc部份),其具有於Asn297上之糖 鏈,其中該糖鏈中之岩藻糖含量為65%或以下(根據Kabat 編號)。於另一實施例中,該糖鏈中之岩藻糖含量為5%至 65%,較佳20%至40%。根據本發明之「Asn297」意指位 於Fc區之約第297位之胺基酸--精胺酸。基於抗體之微小序 列變化,Asn297亦可位於第297位之上游或下游幾個胺基 酸位置處(通常不超過±3個胺基酸位置),亦即位於第294 與第300位之間。於一項實施例中,根據本發明之經醣基 化之抗原結合蛋白的IgG亞類為人類IgGl亞類、具有突變 L234A及L235A之人類IgGl亞類 '或IgG3亞類。於另一實 施例中,N-乙醇醯神經胺糖酸(NGNA)之含量占該糖鏈之 1%或以下、及/或N端α-1,3-半乳糖之含量占1°/。或以下。糖 鏈較佳顯示出如下特徵:與Ν鍵連之聚醣係附接至於CHO 細胞中重組表現之抗體的Asn297處。 術語「糖鏈顯示出如下特徵:與N鍵連之聚醣係附接至 於CHO細胞中重組表現之抗體的Asn297處」係指除岩藻糖 148423.doc •25- 201111394 殘基之外,位於根據本發明之全長母本抗體的Asn297處之 糖鏈的結構及糖殘基序列與在未經修飾之CH〇細胞中表現 之相同抗體(例如於WO 2006/103 100中報導之抗體)相同。 如本申請案所用,術語「NGNA」係指糖殘基--N-乙醇 醯神經胺糖酸。 人類IgGl或IgG3之醣基化發生於Asn297處,其係作為核 心岩藻糖基化之雙觸複合寡糖,且醣基化之末端為至多兩 個Gal殘基。IgG 1或IgG3亞類之人類恆定重鏈區詳細報導 於如下文獻中· Kabat,E·,A.,等人,Sequences of
Proteins of Immunological Interest,第 5 版,Public Health
Service, National Institutes of Health, Bethesda, MD. (1991);及 Brtiggemann,M.,等人,j. Exp. Med. 166 (1987) 1351-1361 ; Love,T.W·,等人,Methods Enzymol. 178(1989)515-527。依末端Gal殘基數量而定,該等結構稱 為 GO、Gl(a-1、6 -或 α-1、3-)、或 G2 聚膽殘基(Raju,t.S., Bioprocess Int. 1(2003)44-53)。抗體 Fc 部份之 CHO 型醣基 化闡述於例如 Routier,F.H·,Glycoconjugate J. 14 (1997)201-207中。於未經糖基修飾之CHO宿主細胞中重組 表現之抗體通常係於Asn297處岩藻糖基化,且其含量為至 少8 5 %。全長母本抗體中之經修飾之寡糖可為雜化型或複 合型。等分、還原/未經岩藻糖基化之寡糖較佳為雜化 型。於另一實施例中’等分、還原/未經岩藻糖基化之寡 糖為複合型。 根據本發明’「岩藻糖含董」意指於糖鍵中Asn297處之 148423.doc -26- 201111394 該種糖之含量,盆rtj ffAA .. 如複合、雜Asn297上之所有糖結構(例 ’、 问甘露糖結構)之總數相關,且ϋ ώ 譜測得,並計算為平均值。岩藻:::: =理之樣本中所判別所有糖結構(例如複合、雜化及 寡甘路糖及高甘露糖結構)的百分比。 藉由重組方法製得根據本發明之抗原結合蛋白。因此, 本發明之第-態樣為—種可編碼根據本發明之抗原結合蛋 白之核I且另-4樣為-種包括該可編碼根據本發明之 抗原結合蛋白之核酸的細胞。重組製備之方法係相關技藝 熟知者’ 1包括於原核及真核細胞中表現蛋白質且隨後 分離出抗原結合蛋白,並通t純化至醫藥上可接受之純 度。爲了於宿主細胞中表現前述抗體,藉由標準方法將分 別編碼經修飾之輕鏈及重鏈的核酸插入表現載體中。於諸 如CH0細胞、NS0細胞、SP2/0細胞、HEK293細胞、COS 細胞、PER.C6細胞、酵母菌或大腸桿菌(E c〇u)細胞之適 宜原核或真核細胞中表現,並自細胞(上清液或溶胞物)收 集抗原結合蛋白。重組產生抗體之一般方法係相關技藝熟 知者’且闡述於例如以下文獻中:Makrides, S.C., Protein
Expr. Purif. 17(1999)183-202 ; Geisse, S·,等人,protein
Expr. Purif. 8(1996)271-282 ; Kaufman, R.J., Mol. Biotechnol. 16(2000)151-161 ; Werner, R.G., Drug Res. 48(1998)870-880。 宜藉由諸如例如蛋白質A-瓊脂糖凝膠術、羥基磷石灰層 148423.doc -27- 201111394 析術、凝膠電泳術、透析術、或親和層析術之習知免疫球 蛋白純化製程,自培養基中分離出根據本發明之雙特異性 抗原結合蛋白。採用常用步驟,很容易分離出編碼單株抗 體的DNA或RNA,並測序。融合瘤細胞可作為該種DNA及 RNA之來源。一旦分離出’即可將DNA插入表現載體中, 隨後轉染至宿主細胞(諸如HEK 293細胞、CHO細胞、或原 本不產生免疫球蛋白之融合瘤細胞)中,使得在宿主細胞 中合成重組單株抗體。 藉由在抗原結合蛋白DNA中引入適宜核苷酸變化、或藉 由核苷酸合成法,製得雙特異性抗原結合蛋白之胺基酸序 列變異體(或突變體然而,僅可於例如如上所述之極有 限之幅度内進行該等修飾。例如,該修飾法不可改變上述 之抗體性質’諸如IgG同型及抗原結合性質,但可改善重 組生產之產量,蛋白質穩定性或便於純化。 如本申请案所用,術語「宿主細胞」係指可經工程處理 產生根據本發明抗體的任一種細胞系統。於一項實施例 中,使用HEK293細胞及CHO細胞作為宿主細胞。如文中 所用,表達方式「細胞」、「細胞系」、及「細胞培養物」 可相互交換使用,且所有該等名稱均包括子代。因此字 詞「轉形體」&「經轉形細胞」包括第一標的細胞及自其 衍生之培養物,而不管轉形物之數量。亦應瞭解由於有 意或無意之變異,所有子代之DNA内容物可能不完全相 同。包括與原始經轉形之細胞具有相同功能或生物學活性 之變異子代。 148423.doc -28· 201111394 於NS0細胞中之表現闡述於例如Barnes, L.M.,等人, Cytotechnology 32(2000)109-123 ; Barnes, L.M.,等人, Biotech. Bioeng. 73(2001)261-270 中。瞬時表現法闡述於 例如 Durocher,Y.,等人,Nucl. Acids. Res. 30(2002)E9 中。可變結構域之選殖法闡述於Orlandi,R.,等人,Proc. Natl. Acad. Sci. USA 86(1989)3833-3837 ; Carter, P.,等 人 ’ Proc. Natl. Acad. Sci. USA 89(1992)4285-4289 ;及 Norderhaug,L·,等人,J. Immunol. Methods 204(1997)77-87中。較佳之瞬時表現系統(Hek 293)闡述於Schlaeger, E.-J.,及 Christensen, K.,Cytotechnology 30(1999)71-83 ;及
Schlaeger, E.-J., J. Immunol. Methods 194(1996)191-199 中。 適於原核生物之對照序列例如包括啟動子、視需要選擇 之操作子序列、及核糖體結合位點。已知真核細胞使用啟 動子、增強子及聚腺苷酸化信號。 當一核酸與另一核酸序列存在功能關係時,稱其「在操 作上相連」。例如’若一前序列或分泌引導序列之DNA表 現出參與多肽分泌之前蛋白,則該DNA係與該多肽iDNA 操作上相連,若啟動子或增強子影響序列之轉錄,則該 文動子或增強子係與該序列在操作上相4 ;或核糖體結合 所處之位置有β於轉冑,則該核糖體位點係與該編碼 序列在操作上相連。「在操作上相連」通常意指所相連之 _序列相鄰接,且若為分泌引導序列,則為相㈣或於 個閱讀框中。缺而辦 …、向增強子並不一定相鄰接。藉由於習 148423.doc -29- 201111394 知之限制酶位點處接合即可完成連接。若不存在該等位 點’則根據習知之操作,使用合成性寡核苷酸接頭或連接 〇 藉由標準技術(包括鹼/SDS處理、CsCl帶狀分離術、管 柱層析術、瓊脂糖凝膠電泳術、及相關技術中已熟知之其 他技術)純化抗體,以除去細胞組分或其他雜質,例如其 他細胞核酸或蛋白質。參見Ausube丨,F,等人,編輯之 Current Protocols in Molecular Biology, Greene Publishing and Wiley Interscience,New York(1987)« 已建立不同方 法,且廣泛應用於蛋白質純化,諸如利用微生物蛋白質之 親和層析術(例如蛋白質A或蛋白質(3親和層析術)、離子交 換層析術(例如陽離子交換(羧甲基樹脂)、陰離子交換(胺 基乙基樹脂)及混合模式交換)、嗜硫吸附術(例如利用卜巯 基乙醇及其他SH配體)、疏水性交互作用或芳香基團吸附 層析術(例如利用苯基-瓊脂糖、親氮雜_芳香系樹脂、或 間-胺基苯基二羥硼酸)、金屬螯合親和層析術(例如利用 Ni(II)-及Cu(Ii)_親和材料)、尺寸排除層析術、及電泳法 (諸如凝膠電泳術、毛細管電泳術)(Vijayalakshmi, M.A.,
Appl. Biochem. Biotech. 75(1998)93-102)。 本發明之一態樣為一#包括根據本發明之抗原結合蛋白 之醫藥組合物。本發明之另一態樣為一種以根據本發明之 抗原結合蛋白於製備醫藥組合物上之用途。本發明之另一 態樣為-種製備包括根據本發明之抗原結合蛋白之醫藥組 合物的方法。於另-態樣中,本發明提供一種組合物,例 148423.doc -30· 201111394 如醫藥組合物,其含有根據本發明之抗原結合蛋白、及與 之調配在一起之醫樂载劑。 本發明之另-態樣為用於治療癌症之該醫藥組合物。 本發明之另-態樣為-種用於治療癌症之根據本發明之 雙特異性抗原結合蛋白。 本發明之另'態樣為-種以根據本發明之抗原結合蛋白 於製備用於治療癌症之醫藥物上之用途。 本發明之另一態樣為一種治療癌症患者之方法其係向 該需要該種治療之患者投與根據本發明之抗原社人蛋白 如文中所用’「醫藥載劑」包括任一種及所有:溶劑: 分散介質、包衣'抗細菌劑及抗真菌劑、等渗及吸收 劑、及生理上相容之類似製劑。較佳地,載劑適於經靜脈 内'經肌内、經皮下、非經腸、經脊髓或經表皮投 如藉由注射或輸液)。 、 可藉由相關技術中已知之多種方法投與本發明組合物。 如熟習此項技術者咸瞭解,投與途徑及/或方式將取決於 所需之結果而變化。爲了經由某也投 、 八 杈樂途偟投與本發明化 ::,可能必須以一種物質包被該化合物、或共同投與該 化^物及一種防止化合物失活 1 M 例如,向受檢者投 /、含於適宜載劑(例如脂質體)或稀釋劑中之化合物。㈣ =受的稀釋劑包括生理食鹽水及水性緩衝溶液。醫藥 、、容液無菌可注射溶液或分散液之無菌水 物質及無菌粉末。該等基質及試劑於醫藥活性 質上之用途係相關技術中已知。 148423.doc •31 · 201111394 如文中所用,紐S吾「非經腸投與」及「非經腸式投與」 意指除了經腸投與及外部投與之外的投與方式通常為經 /主射,且包括但不限於經靜脈内、經肌内、經動脈内、經 鞘内、經被膜内、經眼窩内、經心内、經皮内、經腹膜 内、經氣管、經皮下、經表皮下、經關節内、經被膜下、 經蛛網膜下、經脊柱内及經胸骨内注射及輸液。 如文中所用,術語癌症係指增生性疾病,諸如淋巴瘤、 淋巴細胞性白血病、肺癌、非小細胞肺(NSCL)癌、細支氣 管肺泡細胞肺癌、骨癌、胰腺癌、皮膚癌、頭或頸癌、皮 膚或眼内黑色素瘤、子宮癌、卵巢癌、直腸癌、肛門癌、 月癌(stomach cancer)、胃癌(gastric cancer)、結腸癌、乳 癌、子呂癌、輸卵管癌瘤、子宮内膜癌瘤、子宮頸癌瘤、 陰道癌瘤、外陰癌瘤、霍奇金氏病(H〇dgkin,s Disease)、 食道癌、小腸癌、内分泌系統癌、甲狀腺癌、甲狀旁腺 癌、腎上腺癌、軟組織肉瘤、尿道癌、陰莖癌、前列腺 癌、膀胱癌、腎臟或輸尿管癌、腎細胞癌瘤、腎盂癌瘤、 間皮瘤、肝細胞癌、膽癌、中枢神經系統(CNS)腫瘤、脊 柱(spinal axis)腫瘤、腦幹膠質瘤、多形性膠質母細胞瘤、 星形細胞瘤、許旺氏瘤(schwanomas)、室管膜瘤、髓母細 胞瘤、腦(脊髓)膜瘤、鱗狀細胞癌、垂體腺瘤及依汶氏肉 瘤(Ewings sarcoma) ’包括上述癌症之任何難治療形式, 或一種或多種上述癌症之組合。 該等組合物亦可含有佐劑,諸如防腐劑、濕潤劑、乳化 劑及分散劑。可藉由前述滅菌製程,及藉由包含多種抗細 148423.doc •32. 201111394 菌劑及抗真菌劑,例如對經基苯甲酸顆、氣丁醇、苯齡、 ==等,確保防止微生物之存在。亦可能需要組合物包 =劑,諸如糖'氯化納等。此外,可藉由包含延遲吸 =劑’諸如單硬脂酸铭及明膠’延長可注射醫藥形式之 吸收。 發ΖΓΓ擇之投與途徑,可以適宜水合物形式使用之本 =化5物及/或本發„藥組合物藉由熟習此項技術者 已知之習知方法調配成醫藥上可接受的劑型。 可變化本發明醫藥組合物中之活性成份之實際劑量以 ==份含量可針對特定患者、組合物、及投與方式, 戶無毒性之條件τ,有效獲得所需之治療反應。而 Γ:量將取決於多種藥物動力學因子,包括所使用本 月之特定組合物活性、投與途徑、投與時間 :::物之排泄速率、治療持續時間、與所使用之特= 3物&併使用之其他藥物、化合物及/或物質、接受户療 =者的年齡、性別、體重、狀態、一般健康狀態及先前 病史、及醫學技術中已熟知之類似因素。 ’且°物必須無菌’且其流動程度應可使該組合物可藉由 生理食鹽水溶液。卜,其他較佳之載劑為等渗性緩衝 :例:藉由使用諸如㈣脂之包衣;若為分散液則藉由 ’’所“立徑,及藉由使用界面活性劑維持適宜之 =料多情形t,較佳使組合物包含等滲劑,例如:、 夕元醇諸如甘露醇或山梨糖醇、及氯化納。 14S423.doc -33- 201111394 如文中所用,表達方式Γ細胞」、「細胞株」、及「細胞 培養物」可相互交換使用,且所有該等名稱均包括子代。 因此’字詞「轉形體」及「經轉形之細胞」包括第一標的 細胞及自其衍生之培養物’而不管轉形物之數量。亦應瞭 解,由於有意或無意之突變,所有子代之DNA内容物可以 不70全相同。包括與原始經轉形之細胞具有相同功能或生 物活性之變異子代。當意指確切名稱時,將於上下文中瞭 解。 如文中所用,術語Γ轉形」係指以載體/核酸轉染宿主 細胞之過程。若使用不具有難以穿透之細胞壁壁障之細胞 作為伯主細胞,轉染之方法為例如如Graham及van der Eh, Virology 52(1978)546ff中所述之藉由磷酸鈣沉澱法。然 而,亦可使用諸如核注射或原生質體融合術之向細胞引入 DNA之其他方法。若使用原核細胞或實質上含有細胞壁構 造之細胞,一種轉染方法實例為如c〇hen, S N,等人, pnas 69(1972)211G_2114中所述,制氯化㈣行辦處 理。 如文中所用,「表現」係指核酸轉錄成〇1111^八之過程;及/ 或所轉錄之mRNA(亦稱為轉錄本)隨後轉譯成肽、多肽、 或蛋白質之過程。轉錄本及所編碼之多肽統稱為基因產 物。若聚核苷酸源自基因體DNA,則於真核細胞中之表現 可包括mRNA之剪接。 「載體」為-種核酸分子,特定言之,自我複製之核酸 分子,其將所插入之核酸分子轉染至宿主細胞中及/或於 J48423.doc • 34 · 201111394 宿主細胞之間。該術語包括主要用於將DNA或RN A插入細 胞之載體(例如染色體整合術)、主要用於使DNA或RNA複 製之複製載體、及用於使DNA或RNA轉錄及/或轉譯之表 現載體。亦包括提供一種以上上述功能之載體。 「表現載體」為一種聚核苷酸,當引入適宜宿主細胞 時,其可轉錄並轉譯成多肽。「表現系統」通常係指包括 可用於產生所需表現產物之表現載體的適宜宿主細胞。 提供以下實例、序列表及圖式用於幫助理解本發明,其 確實範圍係於隨附專利申請範圍中出示。應瞭解,在不偏 離本發明之精神下,可修改所出示之製程。 對序列表之闡述 SEQ ID ΝΟ:1 SEQ ID NO:2 SEQ ID NO:3 SEQ ID NO:4 C端與<Ang-2>抗體VH-CH1結構域融合之 經修飾之 <八%-2>抗體重鏈 未經修飾之 <八叩-2>抗體輕鏈 經修飾之<\^0卩>抗體重鏈,其具有CH1-CL交換,且C端係與<VEGF>抗體VH-CL結 構域融合 經修飾之<乂£0卩>抗體輕鏈,其具有CH1-CL交換(<VEGF>抗體VL-CH1結構域) 實例 材料及一般方法 關於人類免疫球蛋白輕鏈及重鏈之核苷酸序列的一般訊 息出示於 Kabat,E.A·,等人,Sequences of Proteins of Immunological Interest,第 5 版,Public Health Service, 148423.doc -35- 201111394
National Institutes of Health,Bethesda,MD(1991)中。根據 EU編號法(Edelman,G.M.,等人,Proc. Natl. Acad. Sci. USA 63(1969)78-85 ; Kabat,E.A.,等人,Sequences of Proteins of Immunological Interest,第 5版,Public Health Service, National Institutes of Health, Bethesda, MD, (1991))對抗體鏈胺基酸編號及命名。 重組DNA技術 如 Sambrook,J.等人,Molecular cloning : A laboratory manual ; Cold Spring Harbor Laboratory Press, Cold Spring Harbor,New York, 1 989中所述,利用標準方法操作DNA。 根據製造商之說明使用分子生物學試劑。 基因合成法 由化學合成法所製得之寡核苷酸製備所需之基因片段。 組裝側接獨特限制性内切酶裂解位點之基因片段的方法 為:藉由包括退火及連接募核苷酸之PCR擴增法’且隨後 經指定之限制酶位點(例如Kpnl/ SacI或AscI/PacI)選殖入 基於 pPCRScript(Stratagene)之 pGA4 選殖載體。藉由 DNA 測序證實經次選殖之基因片段的DNA序列。根據 Geneart(Regensburg,德國)中之分類對基因合成片段進行 排序。 DNA序列測定 藉由於MediGenomix GmbH(Martinsried,德國)或 Sequiserve GmbH(Vaterstetten,德國)中進行雙股測序而決定DNA序 列。 148423.doc -36- 201111394 DNA及蛋白質序列分析及序列數據處理 使用 GCG(Genetics Computer Group,Madison, Wisconsin) 套裝軟體(10.2 版本)及 Infomax's Vector NT1 Advance suite(8.0版本)建立序列、圖譜、分析、註釋及闡述。 表現載體 爲了表現所述之雙特異性四價抗體,使用基於存在或不 存在CMV-Intron A啟動子下之cDNA結構,或基於存在 CMV啟動子下之基因組結構,於細胞(例如HEK293 EBNA 或HEK293-F)中瞬時表現之表現質體變體。 除了抗體表現卡匣之外,載體還包含: -允許該質體於大腸桿菌中複製之複製起點;及 -為大腸桿菌提供胺苄西林(ampicillin)抗性之β-内醯胺 酶基因。 抗體基因之轉錄單位係由如下組件組成: -位於5’端之獨特限制酶位點; -源自人類巨大細胞病毒之中早期增強子及啟動子; _ :¾•為cDNA結構,則隨後接著intron a序列; -人類抗體基因之5,·不轉錄區; -免疫球蛋白重鏈信號序列; -具有免疫球蛋白外顯子-内含子結構之呈cDNA或呈基 因體結構之人類雙特異性四價抗體鏈(野生型或交換 結構域); _具有聚腺苷酸化信號序列之3,不轉錄區;及 -位於3 ’端之獨特限制酶位點。 I48423.doc -37- 201111394 如下所述之包括所述抗體鏈的融合基因的產生方法為: 採用PCR及/或基因合成法,並利用已知之重組方法及技術 組裝’其係例如利用各自載體中之獨特限制酶位點,連接 對應之核S文片#又。藉由DNA測序決定經次選殖之核酸序 列。爲了瞬時轉染’藉由獲自經轉形之大腸桿菌培養物 (Nucleobond AX,Macherey-Nagel)的質體製劑製得大量質 體。 細胞培養技術 採用如下文獻中所述之標準細胞培養技術:Current Protocols in Cell Biology, Bonifacino, J.S., Dasso, M.,
Harford, J.B” Lippincott-Schwartz, J.及 Yamada, K.M.(編 輯),John Wiley & Sons, Inc(2000)。 如下所述,於黏附生長之HEK293-EBNA中,或於懸浮 生長之HEK29-F細胞中’藉由瞬時共轉染各自三種表現質 體,表現雙特異性四價抗體。 於HEK293-EBNA系統中瞬時轉染 取培養於DMEM(經Dulbecco改質之Eagle培養基, Gibco)(其中補充10%極少igG之FCS(胎牛血清,Gibco)、 2 mM L-麵醯胺酸(Gibco)、及 250 pg/ml 遺傳黴素(Gibco)) 中之黏附生長之HEK293-EBNA細胞(表現Epstein-Barr病毒 核抗原之人類胚胎腎臟細胞株293 ;美國菌種收集中心寄 存編號ATCC # CRL-10852,Lot. 959 218)中,分別瞬時共 轉染各三種表現質體(例如編碼經修飾之重鏈、及對應之 輕鏈及經修飾之輕鏈),表現雙特異性四價抗體。爲了轉 148423.doc -38· 201111394 染,依FuGENEtm試劑(μ1)對〇ΝΑ(μ§)比例為4:1(自3:1至 6:1)使用 FiiGENEtm6 轉染試劑(R〇che M〇lecuiar
Biochemicals)。由各自之質體表現蛋白質,且所採用之莫 耳比分別為:編碼(經修飾及野生型)輕鏈的質體對編碼經 修飾之重鏈的質體的莫耳比為1:1:1(等莫耳)、自1:1:2至 2:2:1。於第3天,向細胞添加L-麩醯胺酸4 mM、葡萄糖 [Sigma]及NAA[Gibco]。於轉染之後5至丨丨天,藉由離心, 收集含有雙特異性四價抗體之細胞培養物上清液,並貯存 於-20 C下。關於於例如HEK293細胞中重組表現人類免疫 球蛋白之一般訊息出示於Meissner, P·,等人,Biotechnol, Bioeng. 75(2001)197-203中。 於HEK293-F系統中瞬時表現 或者’可根據製造商之說明,利用HEK293-F系統 (Invitrogen),藉由瞬時轉染各自之質體(例如編碼經修飾 之重鏈的質體、及對應之輕鏈及經修飾之輕鏈的質體)產 生雙特異性四價抗體。簡言之,使HEK293-F細胞 (Invitrogen)懸浮生長於含無血清之FreeStyle 293表現培養 基(Invitrogen)的震盪燒瓶或帶攪拌發酵槽中,並以上述三 種表現質體混合物及293fectin或fectin(Invitrogen)轉染。 於2 L震盪燒瓶(Corning)中,HEK293-F細胞之接種密度為 1.0E*6個細胞/mL,總體積為600 mL,並於120 rpm,8% C〇2下培養。次日,依細胞密度為約1 5E*6個細胞/mL,以 約42 mL含如下物質之混合物轉染細胞:A)含共6〇〇叫質 體 DNA(1 pg/mL)之 20 mL Opti-MEM(Invitrogen),其係依 148423.doc -39- 201111394 等莫耳比例編碼經修飾之重鏈、對應之輕鏈及對應之經修 飾之輕鏈;及B)20 ml Opti-MEM+1.2 mL 293 fectin 或 fectin(2 μΙ/mL)。根據葡萄糖消耗量,於發酵期間添加葡 萄糖溶液。於5至10天之後,收集含有分泌型抗體之上清 液’並直接由上清液純化出抗體’或冷凍並貯存上清液。 蛋白質測定 於280 nm下之光密度(OD)下,利用基於根據pace,CN, 等人,Protein Science,1995, 4, 241 1-1423 中之胺基酸序列 s十算所得之莫耳消光係數,測定已純化之雙特異性四價抗 體及衍生物的蛋白質濃度。 上清液中抗體濃度之測定 利用蛋白質A瓊脂糖小珠(Roche),藉由免疫沉殿術,測 疋細胞培養物上清液中之雙特異性四價抗體的濃度。於 TBS-NP40(50 mM Tris > pH 7.5 > 150 mM NaCl » 1% N〇nidet-P40)中’三次洗滌60 μί蛋白質A瓊脂糖小珠。隨 後’在已先於TBS-NP40中預平衡之蛋白質a瓊脂糖小珠中 添加1至15 mL細胞培養物上清液。於室溫下培養1 h之 後’依如下方式’於Ultrafree-MC-過濾管柱(Amicon)上洗 滌小珠:以0.5 mL TBS-NP40洗滌一次,以〇 5 mL 鱗酸 鹽緩衝液(2xPBS,Roche)洗蘇兩次,且以〇 5 mL 1 00 mM 檸檬酸鈉pH 5,0簡短洗滌四次。藉由添加3 5 μΐ NuPAGE® LDS樣本緩衝液(Invitrogen)溶離出已結合之抗體。取一半 樣本與NuPAGE®樣本還原試劑合併,且另一半保持不還 原,並於70°C下加熱10 min。隨後,將5-30 μΐ添加至4- 148423.doc -40· 201111394 12% NuPAGE® Bis-Tris SDS-PAGE(Invitrogen)(未還原之 SDS-PAGE係使用MOPS緩衝液;且經還原之SDS-PAGE係 使用MES緩衝液及NuPAGE®抗氧化操作緩衝液添加劑 (Invitrogen)),並利用考馬斯藍(Coomassie Blue)染色。 藉由親和HPLC層析術,定量測量細胞培養物上清液中 之雙特異性四價抗體濃度。簡言之,向含200 mM KH2PO4、1〇〇 mM 擦檬酸鈉(pH 7.4)之 Applied Biosystems Poros A/20管柱添加含有已與蛋白質A結合之抗體及衍生 物的細胞培養物上清液,並於AgUent HPLC 11 00系統中, 利用200 mM NaCl、100 mM檸檬酸(pH 2.5),自擔體溶 離。藉由UV吸光度及波峰面積之積分值定量溶離出之蛋 白質。使用經純化之標準IgGl抗體作為標準物。 或者,藉由夾心-IgG-ELISA測定細胞培養物上清液中之 雙特異性四價抗體之濃度。簡言之,依1〇〇 pL/孔,以0.1 pg/mL之經生物素化之抗-人類IgG捕捉分子F(ab')2<h-Fcy> BI(Dianova)塗布 StreptaWell High Bind Strepatavidin A-96 孔微量滴定盤(Roche),於室溫下靜置1 h,或於4°C下靜置 過夜’且隨後以 200 pL/孔之 PBS、〇·〇5% Tween(PBST ’ Sigma)洗務三次。向各孔添加100 μί/孔之含於PBS(Sigma) 中之含各抗體之細胞培養物上清液系列稀釋液,並於室溫 下,於微量滴定盤振盪器中培養1-2 h。以200 pL/孔之 PBST洗滌各孔三次,並利用1〇〇 μΐ 0.1 Mg/mL之 F(ab')2<hFcY>POD(Dianova)作為偵測抗體,於室溫下,於 微量滴定盤振盪器中偵測已結合之抗體1 -2 h。以200 μί/ 148423.doc 41 201111394 孔之PBST洗滌三次,以沖去未結合之偵測抗體,並藉由 添加100 pL ABTS/孔,偵測已結合之偵測抗體。於Tecan Fluor分光計中,於測量波長405 nm下(對照波長492 nm), 測定吸光度。 蛋白質純化 根據標準方法,自經過濾之細胞培養物上清液中純化出 蛋白質。簡言之,在蛋白質A瓊脂糖管柱(GE healthcare) 上添加雙特異性四價抗體,並利用PBS洗滌。於pH 2.8 下,溶離出雙特異性四價抗體,且隨後立即中和樣本。聚 結之蛋白質係於PBS或於20 mM組胺酸、150 mM NaCl(pH 6.0)中經過尺寸排除層析術(Superdex 200,GE Healthcare) 而與單聚抗體分離。彙集單體溶離份,且若需要,則利用 例如 MILLIPORE Amicon Ultra(30 MWCO)離心式濃縮器濃 縮,冷凍並貯存於-20°C或-80°C下。提供部份樣本用於隨 後之蛋白質分析及特徵分析’例如藉由SDS-PAGE、尺寸 排除層析術或質譜。
SDS-PAGE 根據製造商之說明,使用NuPAGE®預鑄凝膠系統(Pre-Cast gel system)(Invitrogen) 。 特 定言之 ,使用 10% 或 4-12%之 NuPAGE® Novex® Bis-TRIS 預鑄凝膠(pH 6.4)、及 NuPAGE® MES(經還原之凝膠,使用NuPAGE®抗氧化操作 緩衝液添加劑)或MOPS(未經還原之凝膠)操作緩衝液。 分析級尺寸排除層析術 藉由HPLC層析術進行尺寸排除層析術,以確定雙特異 148423.doc -42· 201111394 性四價抗體之聚結與寡聚狀態。簡言之,取蛋白質A純化 抗體,在 Agilent HPLC 1100 系統上,於含 300 mM NaCl、 50 mM KH2P〇4/K2HP〇4(pH 7.5)中,加至 Tosoh TSKgel G3000SW管柱,或在Dionex HPLC-系統上,於2xPBS中, 加至Superdex 200管柱(GE Healthcare)中。藉由UV吸光度 及波峰面積之積分值定量溶離出之蛋白質。使用BioRad凝 膠過濾標準151-1901作為標準物。 質譜術
藉由電喷霧離子化質譜(ESI-MS)測定並證實去醣基化之 雙特異性四價抗體之總質量。簡言之,於蛋白質濃度至多 2 mg/ml下,於37°C 下,利用含於 100 mM KH2P04/K2HP04 (pH 7)中之 50 mU N-醣苦酶 F(PNGaseF,ProZyme),使 100 Mg純化抗體進行去醣基化12-24 h,且隨後於Sephadex G25 管柱(GE Healthcare)上進行HPLC而脫鹽化。經去醣基化及 還原之後,藉由ESI-MS測定各經修飾之重鏈、輕鏈及經修 飾之輕鏈之質量。簡言之,由50 pg雙特異性四價抗體於 115 μΐ中,與60 μΐ 1M TCEP及50 μΐ 8 Μ鹽酸胍一起培養, 且隨後脫鹽。藉由配備NanoMate源之Q-Star Elite MS系統 中之ESI-MS測定總質量、及經還原之重鏈及輕鏈之質量。 結合VEGF之ELISA 利用全長 VEGF165-His蛋白質(R&D Systems),於 ELISA 分析中評定雙特異性四價抗體之結合性質。為此,以1 〇〇 μΐ 含2 pg/mL重組人類 VEGF165(R&D Systems)之PBS 溶液 塗布Falcon聚苯乙烯透明加強型微量滴定盤,並靜置於室 148423.doc -43- 201111394 溫下2 h或於4°C下靜置過夜。以300 μΐ PBST(0.2% Tween 20)三次洗滌各孔,並於室溫下,利用200 μΐ 2% BSA 0.1% Tween 20阻斷30 min,且隨後利用300 μΐ PBST洗務三次。 依100 pL/孔,向各孔添加含於PBS(Sigma)中之純化雙特 異性四價抗體之系列稀釋液,並於室溫下,於微量滴定盤 振盪器中,培養1 h。以300 μΐ PBST(0.2% Tween 20)洗滌 各孔三次,並利用100 μΐ^/孔之含於2% BS A 0.1% Tween 20 中之 0.1 pg/ml F(ab') <hFcgamma>POD(Immuno research) 作為偵測抗體,於室溫下,於微量滴定盤振盪器中,偵測 已結合之抗體。以300 μί/孔之PBST洗滌三次,以洗去未 結合之偵測抗體,且藉由添加100 pL ABTS/孔,偵測已結 合之偵測抗體。於Tec an Fluor分光計中,於測量波長405 nm下(參考波長492 nm)測定吸光度。 VEGF結合性:藉由表面電漿共振(Biacore)判定於37°C下 與VEGF結合之動力學特徵 爲了進一步確證ELISA之結果,根據如下方法,利用於 Biacore T1 00儀器上之表面電漿共振術定量分析雙特異性 四價抗體與VEGF之結合,並利用T100套裝軟體進行分 析:簡言之,藉由與山羊-抗-人類IgG(JIR 109-005-098)之 結合,使雙特異性四價抗體捕捉於CM5-晶片上。利用如 下之標準胺基偶合法,藉由胺基偶合,使捕捉抗體固定: 使用HBS-N緩衝液作為操作緩衝液,藉由EDC/NHS混合物 活化,使得配體密度為700 RU。以偶合緩衝液NaAc(pH 5.0)稀釋捕捉抗體,使得c=2 pg/mL,最後注射1 Μ乙醇 148423.doc -44- 201111394 胺,阻斷仍然活化之叛基。於流速5 pL/min下,並依濃度 為c=10 nM,於操作緩衝液+ 1 mg/mL BSA中稀釋,捕捉雙 特異性四價<VEGF>抗體;捕捉量應達到約30 RU。使用 rhVEGF(rhVEGF,R&D-Systems 目錄號 293-VE)作為分析 物。於作為操作緩衝液之PBS+0.005v/v°/〇 Tween20中,於 25°C或37°C下,評估VEGF與雙特異性四價<VEGF>抗體結 合的動力學特性。依流速50 pL/min注入樣本,且與 rhVEGF之自300至0.29 nM的濃度系列的締合時間為80秒 鐘,且解離時間為1200秒鐘。於每一分析物循環之後,使 用10 mM甘胺酸(pH 1.5),並接觸60秒鐘,使游離捕捉抗 體表面再生。利用常用之雙對照方法(對照:rhVEGF與捕 捉分子山羊·抗-人類IgG之結合,以測量中之流槽為背景 值,rhVEGF濃度「0」,模型:一對一朗繆爾結合模型 (Langmuir binding 1:1),(由於係與捕捉分子之結合,將 Rmax設定為局部),計算動力學常數。
結合 Ang-2之 ELISA 利用全長Ang-2-His蛋白質(R&D Systems),於ELISA分 析中評定雙特異性四價抗體之結合性質。為此,以1〇〇 μΐ 含於PBS中之1 pg/mL重組人類Ang-2(R&D Systems,無載 劑)塗布Falcon聚苯乙烯透明加強型微量滴定盤,並於室溫 下靜置2 h,或於4°C下靜置過夜。以300 μΐ PBST(0.2% Tween 20)洗滌各孔三次,並於室溫下,以200 μΐ 2% BSA 0.1% Tween 20阻斷30 min,且隨後以300 μΐ PBST洗滌三 次。向各孔添加100 μί/孔之含於PBS(Sigma)中之純化雙 148423.doc .45- 201111394 特異性四價抗體系列稀釋液,並於室溫下,於微量滴定盤 振盪器上培養1 h。以300 μΐ PBST(0.2% Tween 20)洗務各 孔三次,並利用100 μΐ^/孔之含於2% BSA 0.1% Tween 20中 之 0.1 pg/ml F(ab’)<hk>POD(Biozol 目錄號 206005)作為抗 體,於室溫下,於微量滴定盤振盪器上偵測已結合之抗體 1 h。以300 pL/孔PBST洗滌三次,以洗去未結合之偵測抗 體,並藉由添加100 μί ABTS/孔偵測已結合之偵測抗體。 於Tecan Fluor分光計中,於測量波長405 nm下(對照波長 492 nm),測定吸光度。
結合 Ang-2之 BIACORE 利用 BIACORE T100 儀器(GE Healthcare Biosciences AB,Uppsala,瑞典),藉由表面電漿共振研究雙特異性四 價抗體與人類Ang-2之結合。簡言之,為測定親和力,藉 由胺偶合術,使山羊<hIgG-Fcy>多株抗體固定於CM5晶片 上,以呈現對抗人類Ang-2之雙特異性四價抗體。於25°C 下,於 HBS 緩衝液(HBS-P)(l〇 mM HEPES、150 mM NaCl、0.005°/oTween20(Ph7.4))中,測定結合性。添加不 同浪度之含純化Ang-2-His(R&D Systems或經自行純化)的 溶液。藉由注入Ang-2 3分鐘測定締合度;以HBS緩衝液洗 滌晶片表面3分鐘,測定解離度;並利用一對一朗繆爾結 合模型評估KD值。由於Ang-2製劑之異質性,無法觀察到 1:1結合性;因此KD值僅為相對評估值。自樣本曲線減去 陰性對照數據(例如緩衝液曲線),以校正系統之固有基線 漂移及減少噪音信號。使用Biacore T100評估軟體(版本 148423.doc -46- 201111394 1.1.1)分析感應曲線圖,並計算親和力數據。或者,可藉 由經胺偶合術(無BSA)固定於CM5晶片上之五組胺酸抗體 (PentaHisAntibody)(PentaHis-Ab,無 BSA,Qiagen 第 34660 號)捕捉Ang-2,且捕捉量為2000-1700 RU(參見下文)。
橋連 Ang-2-VEGF之 ELISA 利用經固定之全長VEGF165-His蛋白質(R&D Systems)及 用於偵測已結合之雙特異性抗體的人類Ang-2-His蛋白質 (R&D Systems),於ELIS A分析中評定雙特異性四價抗體之 結合性質。由於只有雙特異性四價<VEGF-Ang-2>抗體能 夠同時結合VEGF及Ang-2,因此其橋連兩種抗原,而單特 異性「標準」IgGl抗體則不能同時結合VEGF及Ang-2(圖 7)。 為此,以100 μΐ之含2 pg/mL重組人類VEGF165(R&D Systems)之PBS溶液塗布Falcon聚苯乙稀透明加強型微量滴 定盤,並於室溫下靜置2 h,或於4°C下靜置過夜。以300 μΐ PBST(0.2% Tween 20)洗滌各孔三次,並於室溫下,以 200 μΐ 2% BSA 0.1% Tween 20阻斷 30 min,且隨後以 300μ1 PBST洗滌三次。向各孔添加100 pL/孔之含於PBS(Sigma) 中之純化雙特異性四價抗體系列稀釋液,並於室溫下,於 微量滴定盤振盪器上培養1 h。以300 μΐ PBST(0.2°/。Tween 20)洗務各孔三次,並藉由添加100 μΐ之含0.5 pg/ml人類 Ang-2-His(R&D Systems)之PBS溶液伯測已結合之抗體。 以300 μΐ PBST(0.2% Tween 20)洗滌各孔三次,並於室溫 下,以 100 μΐ 0.5 pg/mL <Ang‘2>mIgGl-生物素抗體 148423.doc -47- 201111394 (BAM0981,R&D Systems)偵測已結合之 Ang-2 1 h。以 300 μΐ PBST(0.2% Tween 20)洗滌三次,以沖去未結合之偵測 抗體,並藉由添加100 μΐ於封阻緩衝液中稀釋1:4之1:2000 抗生物鏈菌素-POD共輛物(Roche Diagnostics GmbH,目錄 號1 1089153),並置於室溫下lh,偵測已結合之抗體。以 300 μΐ PBST(0.2°/〇 Tween 20)洗滌三至六次,以沖去未結 合之抗生物鏈菌素-POD共軛物,並藉由添加100 μι ABTS/ 孔,偵測已結合之抗生物鏈菌素-POD共軛物。於Tecan Fluor分光計中,於測量波長405 nm下(對照波長492 nm)測 定吸光度。 藉由Biacore證實雙特異性四價之抗體<VEGF-Ang-2>可同 時結合VEGF-A及Ang-2 爲了進一步證實獲自橋連ELISA之數據,進行另一種分 析以證實可同時與VEGF及Ang-2結合,其係根據如下方 法,於Biacore T100儀器上進行表面電漿共振術,並利用 T100 套裝軟體(T100 Control ,2.01 版本;T100 Evaluation,2.01 版本;T100 Kinetics Summary ; 1.01 版 本)分析:於PBS、0.005 v/v°/〇 Tween20操作緩衝液中,利 用藉由胺偶合(BSA-free)固定於CM5晶片上之五組胺酸抗 體(PentaHis-Ab,無 BSA,Qiagen號 34660)捕捉 Ang-2,使 捕捉量為2000-1700 RU。於偶合時,使用HBS-N緩衝液作 為操作緩衝液,藉由EDC/NHS混合物活化。使PentaHis· Ab無BSA捕捉抗體於偶合緩衝液NaAc(pH4.5)中稀釋, c=30 pg/mL,最後注入1 Μ乙醇胺,阻斷仍然活化之羧 148423.doc -48- 201111394 基;測試之配位體密度為5000及17000 RU。於流速5 pL/min下,藉由於操作緩衝液+1 mg/mL BSA中稀釋之 PentaHis-Ab捕捉濃度為 500 nM之Ang-2。隨後,與rhVEGF 一起培養,形成夾心型複合物,證實<Ang-2、VEGF>雙特 異性四價之抗體與Ang-2及VEGF之結合。為此,於流速50 pL/min下,使於操作緩衝液+1 mg/mL BSA中稀釋至濃度 1 00 nM之雙特異性四價之<VEGF-Ang-2>抗體結合至Ang-2,且在流速50 pL/min及VEGF濃度為150 nM下,與含 VEGF(rhVEGF,R&D-Systems 目錄號 293-VE)之 PBS + 0.005 v/v% Tween20操作緩衝液溶液一起培養,彳貞測同時結合 性。締合時間為120秒鐘,解離時間為1200秒鐘。於每一 循環之後,採用流速為50 pL/min之2 X 10 mM甘胺酸(pH 2.0),並使接觸60秒鐘,完成再生。利用常用之雙對照(對 照:雙特異性抗體及rhVEGF與捕捉分子PentaHisAb之結 合)校正感應曲線圖。利用濃度為「0」之rhVEGF測定各 Ab之空白值。 產生HEK293-Tie2細胞株 爲了測定<Ang-2、特異性四價之抗體對細胞上 受Ang-2激發之Tie2填酸化及Ang-2與Tie2之結合的干擾, 故製備重組HEK293-Tie細胞株。簡言之,利用Fugene (Roche Applied Science)作為轉染試劑,使於CMV啟動子 控制下可編碼全長人類Tie2及新黴素(Neomycin)抗性標記 物之基於pcDNA3之質體(RB22-pcDNA3 Topo hTie2)轉染 HEK293 細胞(ATCC),並於 DMEM 10% FCS、500 pg/mi 148423.doc -49- 201111394 G4 1 8中選出具抗性之細胞。利用選殖柱(cloning cylinder) 分離出各細胞株,且隨後藉由FACS分析Tie2表現。鑑定出 第22株系為即使在G418不存在下仍然具有高量且穩定之 Tie2表現量之細胞株(HEK293-Tie2第22株系)。HEK293-Tie2第22株系隨後用於細胞分析:由Ang-2誘發之Tie2磷酸 化及Ang-2細胞配位體結合分析。 由Ang-2誘發之Tie2攝酸化分析 根據如下分析準則,測定<Ang-2、VEGF>雙特異性四價 之抗體對由Ang-2誘發之Tie2磷酸化的抑制。於Ang-2抗體 不存在或存在下,以Ang-2激發HEK293-Tie2第22株系5分 鐘,並藉由夾心ELISA定量P-Tie2。簡言之,於經聚-D-離 胺酸塗布之96孔-微量滴定盤上,依每孔2x1 05個細胞,使 HEK293-Tie2 第 22株系於 100 μΐ DMEM、10% FCS、500 μβ/ηι1遺傳黴素中生長過夜。次曰,於微量滴定盤中製得 一排用於滴定之<Ang-2、VEGF>雙特異性四價之抗體(4倍 濃縮,75 μΐ最終體積/孔,雙重複),並與75 μΐ Ang-2(R&D systems 第 623-AN 號]稀釋液(3.2 pg/ml,4 倍濃縮溶 液)混合。於室溫下,預培養抗體及Ang-2 15 min。向 HEK293-Tie2 第 22 株系細胞(與 1 mM NaV304,Sigma #S6508 —起預培養5 min)添加100 μΐ之該混合物,並於 37°C下培養5 min。隨後,以每孔200 μΐ冰冷之PBS + 1 mM NaV304洗滌細胞,且藉由於冰上,每孔添加120 μΐ之溶胞 緩衝液(20 mM Tris(pH 8.0)、137 mM NaCn、1% ΝΡ-40、 10%甘油、2 mM EDTA、1 mM NaV304、1 mM PMSF及 10 148423.doc -50- 201111394 pg/ml抑肽酶)溶胞。於4°C下,於微量滴定盤振盪器上使 細胞溶解30 min,且在未離心且未測定總蛋白質下,直接 取100 μΐ溶胞產物轉移至p-Tie2丑[18八微量滴定盤(11&0 Systems,R&D #DY990)。根據製造商之說明定量P-Tie2含 量,且利用Excel外掛程式XLfit4(單一位點劑量反應,模 型205)測定抑制性之IC50值。比較實驗内之IC50值,但可 能隨不同實驗之間變化。 由VEGF誘發HUVEC增殖之分析 選擇由VEGF誘發HUVEC(人類臍靜脈上皮細胞, Promocell #C-12200)之增殖用於測定 <Ang-2、VEGF>雙特 異性四價之抗體之細胞功能。簡言之,於經膠原蛋白I塗 布之BD Biocoat膠原蛋白I 96孔微量滴定盤(BD #354407/ 35640)中,使每96個孔5000個HUVEC細胞(低繼代次數, S5次繼代)於100 μΐ飢餓培養基(EBM-2内皮基本培養基 (Promocell # C-22211)、0.5% FCS、青黴素(Penicilline)/鏈 黴素(Streptomycine))中培養過夜。混合不同濃度之<Ang-2、VEGF>雙特異性四價之抗體與rhVEGF(最終濃度30 ngl/ml,BD # 3 54107),並於室溫下預培養15分鐘。隨 後,向HUVEC細胞添加該混合物,並於37°C,5% C02下 培養72 h。於分析當日,使分析板平衡至室溫30 min,並 利用CellTiter-GloTM發光細胞存活率分析套組(Promega,# G7571/2/3),根據手冊測定細胞存活率/增殖。於分光光度 計中測定發光性。 實例1 148423.doc 51 201111394 產生、表現、純化可識別Ang-2及VEGF-Α之雙特異性四 價之抗原結合蛋白、並鑑定其特徵 於第一實例中,製備可識別Ang-2及VEGF-Α之雙特異性 四價之抗原結合蛋白的方法為:經(G4S)4-連接子使可識 別Ang-2之抗體中之VH-CH1結構域融合體與可識別Ang-2 之該抗體之重鏈(SEQ1)的C端融合;且經(G4S)4-連接子, 使可識別VEGF-Α之抗體的VH-CL結構域融合體與可識別 VEGF之CH1-CL交換抗體的重鏈(SEQ3)的C端融合。爲了 誘發兩條不同重鏈之雜二聚化,使SEQ 1含有例如結序列 (T366W),且使SEQ2含有例如穴序列T366S、L368A及 Y407V ,並使其分別含有兩個半胱胺酸殘基 S3 54C/Y349'C,用於在雜二聚化時形成穩定之雙硫橋。爲 了獲得根據本發明之雙特異性四價之抗原結合蛋白,使該 重鏈構築體與可編碼Ang-2抗體之輕鏈(SEQ2)及識別 VEGF-Α之VL-CH1結構域融合體(SEQ4)的質體共同表現。 具結-入-穴修飾之雙特異性四價之抗原結合蛋白的一般結 構圖出示於圖4中。 如一般方法部份中所述,藉由常用分子生物學技術產生 該雙特異性四價之抗原結合蛋白,並如上所述,於 HEK293F細胞中瞬時表現。隨後,藉由蛋白質A親和層析 術並組合尺寸排除層析術,自上清液中純化得到。藉由質 譜鑑定所獲產物之特徵,並分析其性質,諸如經SDS-PAGE測得之純度、單體含量及穩定性。 148423.doc -52- 201111394 表現 純化 效價 [pg/ml] 產生最終產物 均質性(終產物) 9.8 16.7 mg/L 29% 該等數據顯示,可高產量製得該雙特異性四價之抗原結 合蛋白,且其穩定。 隨後,藉由上述ELISA及Biacore分析法探討與Ang-2及 VEGF-A之結合性,及與二者之同時結合性,並分析功能 性質(諸如抑制Tie2磷酸化及抑制由VEGF誘發之HUVEC增 殖),結果顯示,所產生之雙特異性四價抗體能夠與Ang-2 及VEGF-A結合,且同時阻斷其活性。 【圖式簡單說明】 圖1不具CH4結構域之全長抗體的結構圖解,其係與第 一抗原1特異性結合,且具有兩對重鏈及輕鏈,其包括依 通常順序之可變結構域及恆定結構域。 圖2導致不需要之副產物實例的兩種錯誤配對可能性(四 種中之兩種)的結構圖解,其使可與第一抗原1及第二抗原 2結合之雙特異性四價抗體的產量減少,且其中不交換結 構域。 圖3根據本發明之雙特異性四價之抗原結合蛋白的結構 圖解--藉由置換可與第二抗原特異性結合之抗體的重鏈及 輕鏈中之CH1及CL結構域,防止錯誤配對。 圖4具有結及穴之根據本發明之雙特異性四價之抗原結 合蛋白實例的結構圖解。 148423.doc -53- 201111394 序列表 <11〇>瑞士商赫孚孟拉羅股份公司 <120〉雙特異性四價之抗原结合蛋白
<130> 26168 FT <140> 099119745 <141> 2010-06-17 <150> EP09007966.6 <151〉 2009-06-18 <160> 4 <170> Patentln version 3.2 <210〉 1 <211> 711 <212> PRT <213>人工 <220> <223〉C末端與<Ang-2>抗體VH-CH1結構域融合之經修飾之<>\1^-2>抗體重鏈 <400> 1
Gin Val Gin Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ala 15 10 15
Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr 20 25 30
Tyr Met His Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met 35 40 45
Gly Trp lie Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gin Lys Phe 50 55 60
Gin Gly Arg Val Thr Met Thr Arg Asp Thr Ser lie Ser Thr Ala Tyr 65 70 75 80
Met Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys 85 90 95
Ala Arg Ser Pro Asn Pro Tyr Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr 100 105 110 148423·序列表.doc 201111394
Pro Gly Ala Phe Asp lie Trp Gly Gin Gly Thr Met Val Thr Val Ser 115 120 125
Ser Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser 130 135 140
Lys Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp 145 150 155 160
Tyr Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr 165 170 175
Ser Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr 180 185 190
Ser Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin 195 200 205
Thr Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp 210 215 220
Lys Lys Val Glu Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro 225 230 235 240
Cys Pro Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro 245 250 255
Pro Lys Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr 260 265 270
Cys Val Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn 275 280 285
Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg 290 295 300
Glu Glu Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val 305 310 315 320
Leu His Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser 325 330 335 148423-序列表.doc 201111394
Asn Lys Ala Leu Pro Ala Pro lie Glu Lys Thr lie Ser Lys Ala Lys 340 345 350
Gly Gin Pro Arg Glu Pro Gin Val Tyr Thr Leu Pro Pro Cys Arg Asp 355 360 365
Glu Leu Thr Lys Asn Gin Val Ser Leu Trp Cys Leu Val Lys Gly Phe 370 375 380
Tyr Pro Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu 385 390 395 400
Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe 405 410 415
Phe Leu Tyr Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly 420 425 430
Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr 435 440 445
Thr Gin Lys Ser Leu Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser 450 455 460
Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Gin 465 470 475 480
Val Gin Leu Val Glu Ser Gly Ala Glu Val Lys Lys Pro Gly Ala Ser 485 490 495
Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr Tyr 500 505 510
Met His Trp Val Arg Gin Ala Pro Gly Gin Gly Leu Glu Trp Met Gly 515 520 525
Trp lie Asn Pro Asn Ser Gly Gly Thr Asn Tyr Ala Gin Lys Phe Gin 530 535 540
Gly Arg Val Thr Met Thr Arg Asp Thr Ser lie Ser Thr Ala Tyr Met 148423-序列表.doc 201111394 545 550 555 560
Glu Leu Ser Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys Ala 565 570 575
Arg Ser Pro Asn Pro Tyr Tyr Tyr Asp Ser Ser Gly Tyr Tyr Tyr Pro 580 585 590
Gly Ala Phe Asp lie Trp Gly Gin Gly Thr Met Val Thr Val Ser Ser 595 600 605
Ala Ser Thr Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys 610 615 620
Ser Thr Ser Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr 625 630 635 . 640
Phe Pro Glu Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser 645 650 655
Gly Val His Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser 660 665 670
Leu Ser Ser Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr 675 680 685
Tyr lie Cys Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys 690 695 700
Lys Val Glu Pro Lys Ser Cys 705 710 <210> 2 <211> 215 <212> PRT <213〉人工 <220> <223>未經_之<一2>抗體輕鏈 <400> 2
Gin Pro Gly Leu Thr Gin Pro Pro Ser Val Ser Val Ala Pro Gly Gin 15 10 15
Thr Ala Arg lie Thr Cys Gly Gly Asn Asn lie Gly Ser Lys Ser Val -4- 148423-序列表.doc 201111394 20 25 30
His Trp Tyr Gin Gin Lys Pro Gly Gin Ala Pro Val Leu Val Val Tyr 35 40 45
Asp Asp Ser Asp Arg Pro Ser Gly lie Pro Glu Arg Phe Ser Gly Ser 50 55 60
Asn Ser Gly Asn Thr Ala Thr Leu Thr 工le Ser Arg Val Glu Ala Gly 65 70 75 80
Asp Glu Ala Asp Tyr Tyr Cys Gin Val Trp Asp Ser Ser Ser Asp His 85 90 95
Tyr Val Phe Gly Thr Gly Thr Lys Val Thr Val Leu Arg Thr Val Ala 100 105 110
Ala Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser 115 120 125
Gly Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu 130 135 140
Ala Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser 145 150 155 160
Gin Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu 165 170 175
Ser Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val 180 185 190
Tyr Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys 195 200 205
Ser Phe Asn Arg Gly Glu Cys 210 215 <210> 3 <211> 707 <212> PRT <213>人工 <220> <223>具有CHl-CL交換,且C末端與<VEGF>抗體VH-CL結構域融合之經修飾i<VEGF^)L體重鏈 148423-序列表.doc 201111394 <400> 3
Glu Val Gin Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly 1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Asn Tyr 20 25 30
Gly Met Asn Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val 35 40 45
Gly Trp lie Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Ala Asp Phe 50 55 60
Lys Arg Arg Phe Thr Phe Ser Leu Asp Thr Ser Lys Ser Thr Ala Tyr 65 70 75 80
Leu Gin Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys 85 90 95
Ala Lys Tyr Pro His Tyr Tyr Gly Ser Ser His Trp Tyr Phe Asp Val 100 105 110
Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ser Ala Ser Val Ala Ala 115 120 125
Pro Ser Val Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly 130 135 140
Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala 145 150 155 160
Lys Val Gin Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin 165 170 175
Glu Ser Val Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser 180 185 190
Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr 195 200 205
Ala Cys Glu Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys Ser 210 215 220 148423·序列表.doc 201111394
Phe Asn Arg Gly Glu Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro 225 230 235 240
Ala Pro Glu Leu Leu Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys 245 250 255
Pro Lys Asp Thr Leu Met lie Ser Arg Thr Pro Glu Val Thr Cys Val 260 265 270
Val Val Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr 275 280 285
Val Asp Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu 290 295 300
Gin Tyr Asn Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His 305 310 315 320
Gin Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys Lys Val Ser Asn Lys 325 330 335
Ala Leu Pro Ala Pro lie Glu Lys Thr 工le Ser Lys Ala Lys Gly Gin 340 345 350
Pro Arg Glu Pro Gin Val Cys Thr Leu Pro Pro Ser Arg Asp Glu Leu 355 360 365
Thr Lys Asn Gin Val Ser Leu Ser Cys Ala Val Lys Gly Phe Tyr Pro 370 375 380
Ser Asp lie Ala Val Glu Trp Glu Ser Asn Gly Gin Pro Glu Asn Asn 385 390 395 400
Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu 405 410 415
Val Ser Lys Leu Thr Val Asp Lys Ser Arg Trp Gin Gin Gly Asn Val 420 425 430
Phe Ser Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gin 435 440 445
Lys Ser Leu Ser Leu Ser Pro Gly Lys Gly Gly Gly Gly Ser Gly Gly 148423-序列表.doc 201111394 450 455 460
Gly Gly Ser Gly Gly Gly Gly Ser Gly Gly Gly Gly Ser Glu Val Gin 465 470 475 480
Leu Val Glu Ser Gly Gly Gly Leu Val Gin Pro Gly Gly Ser Leu Arg 485 490 495
Leu Ser Cys Ala Ala Ser Gly Tyr Thr Phe Thr Asn Tyr Gly Met Asn 500 505 510
Trp Val Arg Gin Ala Pro Gly Lys Gly Leu Glu Trp Val Gly Trp lie 515 520 525
Asn Thr Tyr Thr Gly Glu Pro Thr Tyr Ala Ala Asp Phe Lys Arg Arg 530 535 540
Phe Thr Phe Ser Leu Asp Thr Ser Lys Ser Thr Ala Tyr Leu Gin Met 545 550 555 560
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Lys Tyr 565 570 575
Pro His Tyr Tyr Gly Ser Ser His Trp Tyr Phe Asp Val Trp Gly Gin 580 585 590
Gly Thr Leu Val Thr Val Ser Ser Ala Ser Val Ala Ala Pro Ser Val 595 600 605
Phe lie Phe Pro Pro Ser Asp Glu Gin Leu Lys Ser Gly Thr Ala Ser 610 615 620
Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala Lys Val Gin 625 630 635 640
Trp Lys Val Asp Asn Ala Leu Gin Ser Gly Asn Ser Gin Glu Ser Val 645 650 655
Thr Glu Gin Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser Ser Thr Leu 660 665 670
Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr Ala Cys Glu 675 680 685 148423-序列表.doc 201111394
Val Thr His Gin Gly Leu Ser Ser Pro Val Thr Lys Ser Phe Asn Arg 690 695 700
Gly Glu Cys 705 <210> 4 <211> 212 <212> PRT <213〉人工 <220> <223>具有CH1-CL交換(<VEGF>iSJt VL-CH1結構域)之經修飾之<VEGFH?L體輕鏈 <400> 4
Asp He Gin Met Thr Gin Ser Pro Ser Ser Leu Ser Ala Ser Val Gly 15 10 15
Asp Arg Val Thr lie Thr Cys Ser Ala Ser Gin Asp lie Ser Asn Tyr 20 25 30
Leu Asn Trp Tyr Gin Gin Lys Pro Gly Lys Ala Pro Lys Val Leu lie 35 40 45
Tyr Phe Thr Ser Ser Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly 50 55 60
Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr lie Ser Ser Leu Gin Pro 65 70 75 80
Glu Asp Phe Ala Thr Tyr Tyr Cys Gin Gin Tyr Ser Thr Val Pro Trp 85 90 95
Thr Phe Gly Gin Gly Thr Lys Val Glu lie Lys Ser Ser Ala Ser Thr 100 105 110
Lys Gly Pro Ser Val Phe Pro Leu Ala Pro Ser Ser Lys Ser Thr Ser 115 120 125
Gly Gly Thr Ala Ala Leu Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu 130 135 140
Pro Val Thr Val Ser Trp Asn Ser Gly Ala Leu Thr Ser Gly Val His 145 150 155 160 148423-序列表doc 201111394
Thr Phe Pro Ala Val Leu Gin Ser Ser Gly Leu Tyr Ser Leu Ser Ser 165 170 175
Val Val Thr Val Pro Ser Ser Ser Leu Gly Thr Gin Thr Tyr lie Cys 180 185 190
Asn Val Asn His Lys Pro Ser Asn Thr Lys Val Asp Lys Lys Val Glu 195 200 205
Pro Lys Ser Cys 210 10· 148423·序列表.doc
Claims (1)
- 201111394 七、申請專利範圍: 1 · 一種雙特異性四價之抗原結合蛋白質,其包含: a) 第一抗體之經修獅重鍵, 其與第一抗原特異性結合, 且该重鍵之C端另與該第一抗體之VH-CH1結構域的N - 端融合; b) 如a)之該第一抗體之兩條輕鏈; c) 第一 ·Ι/ι*體之經修飾重鏈, 其與第二抗原特異性結合, 其中CH1結構域經該第二抗體之CL結構域置換, 且該重鏈之C端另與該第二抗體之VH_CL結構域的^^端 融合;及 d) 如c)之β玄第一抗體之兩條經修飾之輕鍵, 其中CL結構域經該第二抗體之CH1結構域置換。 2·如请求項1之抗原結合蛋白,其特徵為: a)之抗體之經修飾重鏈的CH3結構域與c)之抗體之經修 飾重鏈的CH3結構域各於一個包含該等抗體CH3結構 域之間原始介面之介面會合(meet); . 其中該介面經改變以促進形成該雙特異性四價之抗原 結合蛋白’其中該改變之特徵為: i) 一條重鍵之CH3結構域改變, 使得在該雙特異性四價之抗原結合蛋白中與另一條 重鍵之CH3結構域之原始介面會合之一條重鍵之 CH3結構域之原始介面内, 148423.doc 201111394 一個胺基酸殘基被一個具有較大側鏈體積之胺基酸 殘基置換’由此在一條重鏈之CH3結構域之介面内 產生一個突起’其可置於另一條重鏈之CH3結構域 之介面内一個孔穴中 及 ii)另一條重鍵之CH3結構域改變, 使得在該雙特異性四價之抗原結合蛋白中與該第一 CH3結構域之原始介面會合之該第二CH3結構域之 原始介面内, 一個胺基酸殘基被一個具有較小側鏈體積之胺基酸 殘基置換,由此於該第二CH3結構域之介面内產生 一個孔穴,其中可置入該第一 CH3結構域之介面内 之一個突起。 3·如請求項2之抗原結合蛋白,其特徵為: 該具有較大側鏈體積之胺基酸殘基係選自由以下組成 之群:精胺酸(R)、苯丙胺酸(F)、赂胺酸(γ)、色胺酸 (W),及 該具有較小侧鏈體積之胺基酸殘基係選自由以下組成 之群:丙胺酸(A)、絲胺酸(S)、蘇胺酸(τ)、纈胺酸(v)。 4.如請求項2或3之抗原結合蛋白,其特徵為: 兩個CH3結構域進一步改變,藉由半胱胺酸(c)胺基酸 引入各CH3結構域之對應位置’使得兩個CH3結構域之 間可形成雙硫橋。 5·—種製備如請求項1至4之雙特異性四價之抗原結合蛋白 148423.doc -2- 201111394 的方法,b)該宿主細 培養;及 胞於允許合成該抗原 至4之雙特異性四價之抗原結合 轉形宿主細皰; 結合蛋白分子之條件下 c)自該培養物回收該抗原結合蛋白分子。 6. 一種包含如請求項5之載體的宿主細胞。 7· 一種醫藥組合物,其包含如請求項⑴之雙特異性四價 之抗原結合蛋白及至少一種醫藥上可接受之賦形劑。 8.如請求項1至4之雙特異性四價之抗原結合蛋白其係用 於治療癌症。 9 · 種如睛求項1至4之雙特異性四價之抗原結合蛋白於製 備用於治療癌症之藥物之用途。 148423.doc
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
EP09007966 | 2009-06-18 |
Publications (1)
Publication Number | Publication Date |
---|---|
TW201111394A true TW201111394A (en) | 2011-04-01 |
Family
ID=41168686
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
TW099119745A TW201111394A (en) | 2009-06-18 | 2010-06-17 | Bispecific, tetravalent antigen binding proteins |
Country Status (20)
Country | Link |
---|---|
US (2) | US8703132B2 (zh) |
EP (1) | EP2443153B1 (zh) |
JP (1) | JP5689118B2 (zh) |
KR (1) | KR101431344B1 (zh) |
CN (1) | CN102803295B (zh) |
AR (1) | AR077111A1 (zh) |
AU (1) | AU2010262130A1 (zh) |
BR (1) | BRPI1010109A2 (zh) |
CA (1) | CA2764393C (zh) |
CL (1) | CL2011003149A1 (zh) |
ES (1) | ES2442917T3 (zh) |
HK (1) | HK1174645A1 (zh) |
IL (1) | IL216620A0 (zh) |
MX (1) | MX2011013616A (zh) |
PE (1) | PE20120414A1 (zh) |
RU (1) | RU2604189C2 (zh) |
SG (1) | SG177272A1 (zh) |
TW (1) | TW201111394A (zh) |
WO (1) | WO2010145793A1 (zh) |
ZA (1) | ZA201108922B (zh) |
Families Citing this family (187)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7658924B2 (en) * | 2001-10-11 | 2010-02-09 | Amgen Inc. | Angiopoietin-2 specific binding agents |
US20090162359A1 (en) | 2007-12-21 | 2009-06-25 | Christian Klein | Bivalent, bispecific antibodies |
US9266967B2 (en) * | 2007-12-21 | 2016-02-23 | Hoffmann-La Roche, Inc. | Bivalent, bispecific antibodies |
NZ586701A (en) | 2008-01-03 | 2013-07-26 | Scripps Research Inst | Antibody targeting through a modular recognition domain (MRD) wherein the MRD targets angiopoietin-2 (ANG-2) |
US8557243B2 (en) | 2008-01-03 | 2013-10-15 | The Scripps Research Institute | EFGR antibodies comprising modular recognition domains |
US8454960B2 (en) | 2008-01-03 | 2013-06-04 | The Scripps Research Institute | Multispecific antibody targeting and multivalency through modular recognition domains |
US8557242B2 (en) | 2008-01-03 | 2013-10-15 | The Scripps Research Institute | ERBB2 antibodies comprising modular recognition domains |
US8574577B2 (en) | 2008-01-03 | 2013-11-05 | The Scripps Research Institute | VEGF antibodies comprising modular recognition domains |
US8133979B2 (en) * | 2008-12-16 | 2012-03-13 | Hoffmann-La Roche Inc. | Antibodies against human angiopoietin 2 |
MX2011010159A (es) | 2009-04-02 | 2011-10-17 | Roche Glycart Ag | Anticuerpos multiespecificos que comprenden anticuerpos de longitud completa y fragmentos fab de cadena sencilla. |
JP5616428B2 (ja) | 2009-04-07 | 2014-10-29 | ロシュ グリクアート アクチェンゲゼルシャフト | 三価の二重特異性抗体 |
EP2435473B1 (en) | 2009-05-27 | 2013-10-02 | F.Hoffmann-La Roche Ag | Tri- or tetraspecific antibodies |
US9676845B2 (en) | 2009-06-16 | 2017-06-13 | Hoffmann-La Roche, Inc. | Bispecific antigen binding proteins |
US8703132B2 (en) | 2009-06-18 | 2014-04-22 | Hoffmann-La Roche, Inc. | Bispecific, tetravalent antigen binding proteins |
WO2011034605A2 (en) | 2009-09-16 | 2011-03-24 | Genentech, Inc. | Coiled coil and/or tether containing protein complexes and uses thereof |
AR080793A1 (es) | 2010-03-26 | 2012-05-09 | Roche Glycart Ag | Anticuerpos biespecificos |
WO2011147834A1 (en) | 2010-05-26 | 2011-12-01 | Roche Glycart Ag | Antibodies against cd19 and uses thereof |
US20120100166A1 (en) | 2010-07-15 | 2012-04-26 | Zyngenia, Inc. | Ang-2 Binding Complexes and Uses Thereof |
EP2824457A1 (en) | 2010-07-19 | 2015-01-14 | F. Hoffmann-La Roche AG | Method to identify a patient with an increased likelihood of responding to an anti-cancer therapy |
MX2013000667A (es) | 2010-07-19 | 2013-02-27 | Hoffmann La Roche | Metodo para identificar pacientes con probabilidad incrementada de responder a una terapia anticancer. |
WO2012010582A1 (en) | 2010-07-21 | 2012-01-26 | Roche Glycart Ag | Anti-cxcr5 antibodies and methods of use |
CA2805564A1 (en) | 2010-08-05 | 2012-02-09 | Stefan Jenewein | Anti-mhc antibody anti-viral cytokine fusion protein |
CN103068846B9 (zh) | 2010-08-24 | 2016-09-28 | 弗·哈夫曼-拉罗切有限公司 | 包含二硫键稳定性Fv片段的双特异性抗体 |
RS59589B1 (sr) | 2010-11-05 | 2019-12-31 | Zymeworks Inc | Dizajniranje stabilnog heterodimernog antitela sa mutacijama u fc domenu |
WO2012085111A1 (en) | 2010-12-23 | 2012-06-28 | F. Hoffmann-La Roche Ag | Polypeptide-polynucleotide-complex and its use in targeted effector moiety delivery |
AR085403A1 (es) | 2011-02-28 | 2013-09-25 | Hoffmann La Roche | Proteinas monovalentes que se unen a antigenos |
WO2012116926A1 (en) * | 2011-02-28 | 2012-09-07 | F. Hoffmann-La Roche Ag | Antigen binding proteins |
EA201892619A1 (ru) * | 2011-04-29 | 2019-04-30 | Роше Гликарт Аг | Иммуноконъюгаты, содержащие мутантные полипептиды интерлейкина-2 |
CN106432506A (zh) | 2011-05-24 | 2017-02-22 | 泽恩格尼亚股份有限公司 | 多价和单价多特异性复合物及其用途 |
BR112013032235A2 (pt) | 2011-06-15 | 2016-11-22 | Hoffmann La Roche | anticorpos do receptor de epo anti-humano e métodos de uso |
AR086982A1 (es) | 2011-06-22 | 2014-02-05 | Hoffmann La Roche | Eliminacion de celulas diana por parte de celulas t citotoxicas especificas de virus utilizando complejos que comprenden mhc de clase i |
EP2543680A1 (en) * | 2011-07-07 | 2013-01-09 | Centre National de la Recherche Scientifique | Multispecific mutated antibody Fab fragments |
AU2012288846B2 (en) * | 2011-07-27 | 2016-05-19 | Glaxo Group Limited | Anti-VEGF single variable domains fused to fc domains |
CA2837975C (en) * | 2011-08-23 | 2022-04-05 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
PL2748202T3 (pl) * | 2011-08-23 | 2018-12-31 | Roche Glycart Ag | Dwuswoiste cząsteczki wiążące antygen |
US20130078250A1 (en) * | 2011-08-23 | 2013-03-28 | Oliver Ast | Bispecific t cell activating antigen binding molecules |
CN107586340B (zh) | 2011-08-23 | 2022-01-21 | 罗切格利卡特公司 | 对t细胞活化性抗原和肿瘤抗原特异性的双特异性抗体及使用方法 |
RU2014114119A (ru) | 2011-09-23 | 2015-10-27 | Рош Гликарт Аг | Биспецифические анти-egfr/анти igf-1r-антитела |
CN104080811B (zh) | 2011-11-04 | 2019-09-27 | 酵活有限公司 | 在Fc结构域中具有突变的稳定异源二聚的抗体设计 |
BR112014011535A2 (pt) | 2011-12-19 | 2017-05-09 | Hoffmann La Roche | método para a determinação in vitro e imunológica de presença da presença e/ou da quantidade de um parceiro de ligação, método in vitro e utilização de um anticorpo |
CA3149402A1 (en) | 2011-12-22 | 2013-06-27 | F. Hoffman-La Roche Ag | Expression vector element combinations, novel production cell generation methods and their use for the recombinant production of polypeptides |
CN104011080B (zh) | 2011-12-22 | 2017-10-20 | 弗·哈夫曼-拉罗切有限公司 | 用于真核细胞的全长抗体展示系统及其用途 |
KR102080356B1 (ko) | 2011-12-22 | 2020-02-24 | 에프. 호프만-라 로슈 아게 | 발현 벡터 구성, 신규한 생산 세포 생성 방법 및 폴리펩티드의 재조합 생산을 위한 그의 용도 |
CN104105966B (zh) | 2012-02-01 | 2016-10-26 | 弗·哈夫曼-拉罗切有限公司 | 用于检测多特异性结合物的结合搭档的方法 |
JP6363021B2 (ja) | 2012-02-03 | 2018-07-25 | エフ・ホフマン−ラ・ロシュ・アクチェンゲゼルシャフト | 抗原をトランスフェクトされたt細胞を伴う二重特異性抗体分子及び医薬におけるそれらの使用 |
RU2644341C2 (ru) | 2012-02-10 | 2018-02-08 | Дженентек, Инк. | Одноцепочечные антитела и другие гетеромультимеры |
MX360352B (es) | 2012-02-15 | 2018-10-30 | Hoffmann La Roche | Cromatografia de afinidad basada en receptores fc. |
KR101674784B1 (ko) | 2012-04-05 | 2016-11-09 | 에프. 호프만-라 로슈 아게 | 인간 tweak 및 인간 il17에 대한 이중특이적 항체 및 이의 용도 |
US9499634B2 (en) | 2012-06-25 | 2016-11-22 | Zymeworks Inc. | Process and methods for efficient manufacturing of highly pure asymmetric antibodies in mammalian cells |
EP2867253B1 (en) | 2012-06-27 | 2016-09-14 | F. Hoffmann-La Roche AG | Method for selection and production of tailor-made highly selective and multi-specific targeting entities containing at least two different binding entities and uses thereof |
CA2871882A1 (en) | 2012-06-27 | 2014-01-03 | F. Hoffmann-La Roche Ag | Method for making antibody fc-region conjugates comprising at least one binding entity that specifically binds to a target and uses thereof |
HUE029435T2 (en) | 2012-07-04 | 2017-02-28 | Hoffmann La Roche | Anti-theophylline antibodies and methods of their application |
EP3339328A1 (en) | 2012-07-04 | 2018-06-27 | F. Hoffmann-La Roche AG | Anti-biotin antibodies and methods of use |
CN107973856B (zh) | 2012-07-04 | 2021-11-23 | 弗·哈夫曼-拉罗切有限公司 | 共价连接的抗原-抗体缀合物 |
ES2627482T3 (es) | 2012-07-13 | 2017-07-28 | F. Hoffmann-La Roche Ag | Procedimiento para la detección de una proteína de unión multiespecífica |
RU2646159C2 (ru) | 2012-09-14 | 2018-03-01 | Ф. Хоффманн-Ля Рош Аг | Способ получения и отбора молекул, включающих по меньшей мере две различные группировки, и их применение |
JP6581505B2 (ja) * | 2012-10-03 | 2019-09-25 | ザイムワークス,インコーポレイテッド | 重鎖および軽鎖ポリペプチドの対を定量化する方法 |
EP2905290B1 (en) | 2012-10-05 | 2019-12-04 | Kyowa Kirin Co., Ltd. | Heterodimeric protein composition |
KR20150064068A (ko) * | 2012-10-08 | 2015-06-10 | 로슈 글리카트 아게 | 2개의 Fab 단편을 포함하는 FC-부재 항체 및 이용 방법 |
EP2917243B1 (en) | 2012-11-08 | 2018-03-14 | F.Hoffmann-La Roche Ag | Her3 antigen binding proteins binding to the beta-hairpin of her3 |
US9914785B2 (en) | 2012-11-28 | 2018-03-13 | Zymeworks Inc. | Engineered immunoglobulin heavy chain-light chain pairs and uses thereof |
CA2893562C (en) | 2012-11-28 | 2023-09-12 | Zymeworks Inc. | Engineered immunoglobulin heavy chain-light chain pairs and uses thereof |
CN105143270B (zh) | 2013-02-26 | 2019-11-12 | 罗切格利卡特公司 | 双特异性t细胞活化抗原结合分子 |
MY192312A (en) | 2013-02-26 | 2022-08-17 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
EA201890895A1 (ru) | 2013-03-15 | 2019-02-28 | Зинджения, Инк. | Мультивалентные и моновалентные мультиспецифические комплексы и их применение |
WO2014144357A1 (en) | 2013-03-15 | 2014-09-18 | Merck Patent Gmbh | Tetravalent bispecific antibodies |
CA2904805A1 (en) | 2013-04-29 | 2014-11-06 | F. Hoffmann-La Roche Ag | Fc-receptor binding modified asymmetric antibodies and methods of use |
EP3327034A1 (en) | 2013-04-29 | 2018-05-30 | F. Hoffmann-La Roche AG | Fcrn-binding abolished anti-igf-1r antibodies and their use in the treatment of vascular eye diseases |
KR20210094669A (ko) | 2013-04-29 | 2021-07-29 | 에프. 호프만-라 로슈 아게 | 인간 fcrn-결합 변형된 항체 및 사용 방법 |
WO2015025054A1 (en) | 2013-08-22 | 2015-02-26 | Medizinische Universität Wien | Dye-specific antibodies for prestained molecular weight markers and methods producing the same |
CA2922912A1 (en) * | 2013-10-11 | 2015-04-16 | F. Hoffmann-La Roche Ag | Multispecific domain exchanged common variable light chain antibodies |
GB2519786A (en) * | 2013-10-30 | 2015-05-06 | Sergej Michailovic Kiprijanov | Multivalent antigen-binding protein molecules |
EP3783020A1 (en) | 2013-11-21 | 2021-02-24 | F. Hoffmann-La Roche AG | Anti-alpha-synuclein antibodies and methods of use |
AU2014357292B2 (en) | 2013-11-27 | 2020-06-25 | Zymeworks Bc Inc. | Bispecific antigen-binding constructs targeting HER2 |
CA2932958A1 (en) | 2013-12-20 | 2015-06-25 | F. Hoffmann-La Roche Ag | Humanized anti-tau(ps422) antibodies and methods of use |
PL3089996T3 (pl) | 2014-01-03 | 2021-12-13 | F. Hoffmann-La Roche Ag | Dwuswoiste przeciwciała przeciw haptenowi/przeciw receptorowi występującemu w barierze krew-mózg, ich kompleksy i ich zastosowanie jako przenośniki wahadłowe występujące w barierze krew-mózg |
CA2930046A1 (en) | 2014-01-03 | 2015-07-09 | F. Hoffmann-La Roche Ag | Covalently linked polypeptide toxin-antibody conjugates |
MX2016008189A (es) | 2014-01-03 | 2016-09-29 | Hoffmann La Roche | Conjugados helicoidales-anticuerpo anti-helicoidal unidos covalentemente y usos de los mismos. |
KR102381685B1 (ko) | 2014-01-06 | 2022-04-01 | 에프. 호프만-라 로슈 아게 | 1가 혈액 뇌 장벽 셔틀 모듈 |
CN110903398B (zh) | 2014-01-15 | 2023-08-15 | 豪夫迈·罗氏有限公司 | 具有修饰的FCRN和保持的蛋白A结合性质的Fc区变体 |
EP3126389A1 (en) | 2014-04-02 | 2017-02-08 | F. Hoffmann-La Roche AG | Method for detecting multispecific antibody light chain mispairing |
EP3107938B1 (en) | 2014-05-28 | 2022-05-04 | Zymeworks Inc. | Modified antigen binding polypeptide constructs and uses thereof |
GB201411320D0 (en) | 2014-06-25 | 2014-08-06 | Ucb Biopharma Sprl | Antibody construct |
AR100978A1 (es) | 2014-06-26 | 2016-11-16 | Hoffmann La Roche | LANZADERAS CEREBRALES DE ANTICUERPO HUMANIZADO ANTI-Tau(pS422) Y USOS DE LAS MISMAS |
WO2015197736A1 (en) | 2014-06-26 | 2015-12-30 | F. Hoffmann-La Roche Ag | Anti-brdu antibodies and methods of use |
US9840553B2 (en) | 2014-06-28 | 2017-12-12 | Kodiak Sciences Inc. | Dual PDGF/VEGF antagonists |
TW201623329A (zh) | 2014-06-30 | 2016-07-01 | 亞佛瑞司股份有限公司 | 針對骨調素截斷變異體的疫苗及單株抗體暨其用途 |
WO2016001485A1 (en) | 2014-06-30 | 2016-01-07 | Glykos Finland Oy | Saccharide derivative of a toxic payload and antibody conjugates thereof |
JP2017520575A (ja) | 2014-07-01 | 2017-07-27 | ファイザー・インク | 二重特異的ヘテロ二量体ダイアボディおよびその使用 |
CN107074925B (zh) | 2014-07-10 | 2021-08-24 | 阿费里斯股份公司 | 用于预防和/或治疗亨廷顿氏病的物质和方法 |
EP3174897B1 (en) * | 2014-07-29 | 2020-02-12 | F.Hoffmann-La Roche Ag | Multispecific antibodies |
DK3608337T3 (da) | 2014-08-04 | 2024-06-17 | Hoffmann La Roche | Bispecifikke T-celleaktiverende antigenbindende molekyler |
MA41044A (fr) | 2014-10-08 | 2017-08-15 | Novartis Ag | Compositions et procédés d'utilisation pour une réponse immunitaire accrue et traitement contre le cancer |
JP6576456B2 (ja) * | 2014-11-06 | 2019-09-18 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | 修飾されたFcRn結合特性およびプロテインA結合特性を有するFc領域変種 |
PL3215528T3 (pl) | 2014-11-06 | 2020-01-31 | F.Hoffmann-La Roche Ag | Warianty regionu Fc ze zmodyfikowanym wiązaniem FcRn i sposoby stosowania |
PL3221355T3 (pl) | 2014-11-20 | 2021-03-08 | F. Hoffmann-La Roche Ag | Terapia skojarzona składająca się z dwuswoistych aktywujących limfocyty T cząsteczek wiążących antygen CD3 i receptor folianowy 1 (FolR1) oraz antagonistów wiązania osi PD-1 |
EP3227341A1 (en) | 2014-12-02 | 2017-10-11 | CeMM - Forschungszentrum für Molekulare Medizin GmbH | Anti-mutant calreticulin antibodies and their use in the diagnosis and therapy of myeloid malignancies |
CN107001482B (zh) | 2014-12-03 | 2021-06-15 | 豪夫迈·罗氏有限公司 | 多特异性抗体 |
BR112017011170A2 (pt) | 2014-12-18 | 2018-02-27 | Hoffmann La Roche | método para determinar a citotoxicidade dependente do complemento de uma composição |
AR103477A1 (es) | 2015-01-28 | 2017-05-10 | Lilly Co Eli | Compuestos de vegfa / ang2 |
PL3313877T3 (pl) | 2015-06-24 | 2020-11-02 | F. Hoffmann-La Roche Ag | Humanizowane przeciwciała przeciwko białku tau(pS422) i sposoby stosowania |
AU2016323440B2 (en) | 2015-09-15 | 2023-07-13 | Amgen Inc. | Tetravalent bispecific and tetraspecific antigen binding proteins and uses thereof |
AR106188A1 (es) | 2015-10-01 | 2017-12-20 | Hoffmann La Roche | Anticuerpos anti-cd19 humano humanizados y métodos de utilización |
EP3356406A1 (en) * | 2015-10-02 | 2018-08-08 | H. Hoffnabb-La Roche Ag | Bispecific anti-human cd20/human transferrin receptor antibodies and methods of use |
MX2018003629A (es) | 2015-10-02 | 2018-08-01 | Hoffmann La Roche | Anticuerpos anti-pd1 y metodos de uso. |
EP3150636A1 (en) * | 2015-10-02 | 2017-04-05 | F. Hoffmann-La Roche AG | Tetravalent multispecific antibodies |
CR20180162A (es) | 2015-10-02 | 2018-05-25 | Hoffmann La Roche | Moléculas biespecíficas de unión a antígeno activadoras de células t |
WO2017055399A1 (en) | 2015-10-02 | 2017-04-06 | F. Hoffmann-La Roche Ag | Cellular based fret assay for the determination of simultaneous binding |
JP7074665B2 (ja) * | 2015-10-07 | 2022-05-24 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | 共刺激tnf受容体に対する四価の二重特異性抗体発明の分野 |
IL295756A (en) | 2015-10-29 | 2022-10-01 | Hoffmann La Roche | Antibodies against fc-variable region and methods of use |
EP3184547A1 (en) | 2015-10-29 | 2017-06-28 | F. Hoffmann-La Roche AG | Anti-tpbg antibodies and methods of use |
CA2997406C (en) | 2015-12-09 | 2024-05-28 | F. Hoffmann-La Roche Ag | Type ii anti-cd20 antibody for reducing formation of anti-drug antibodies or cytokine release |
CR20180365A (es) | 2015-12-16 | 2018-09-28 | Amgen Inc | PROTEÍNAS DE UNIÓN AL ANTÍGENO BISPECÍFICO DE ANTI-TL1A/ANTI-TNF-a Y SUS USOS |
AU2016381964B2 (en) | 2015-12-30 | 2024-02-15 | Kodiak Sciences Inc. | Antibodies and conjugates thereof |
AU2017205089B2 (en) | 2016-01-08 | 2023-10-05 | F. Hoffmann-La Roche Ag | Methods of treating CEA-positive cancers using PD-1 axis binding antagonists and anti-CEA/anti-CD3 bispecific antibodies |
EP3433281A1 (en) | 2016-03-21 | 2019-01-30 | Elstar Therapeutics, Inc. | Multispecific and multifunctional molecules and uses thereof |
DK3433280T3 (da) | 2016-03-22 | 2023-06-19 | Hoffmann La Roche | Protease-aktiverede T-celle-bispecifikke molekyler |
MX2018015173A (es) | 2016-06-17 | 2019-07-04 | Genentech Inc | Purificacion de anticuerpos multiespecificos. |
US20190233534A1 (en) | 2016-07-14 | 2019-08-01 | Fred Hutchinson Cancer Research Center | Multiple bi-specific binding domain constructs with different epitope binding to treat cancer |
CN116731197A (zh) | 2016-09-19 | 2023-09-12 | 豪夫迈·罗氏有限公司 | 基于补体因子的亲和层析 |
JP6785372B2 (ja) | 2016-09-30 | 2020-11-18 | エフ.ホフマン−ラ ロシュ アーゲーF. Hoffmann−La Roche Aktiengesellschaft | 多重特異性分子の機能分析のためのsprに基づく二重結合アッセイ |
JP7022123B2 (ja) | 2016-09-30 | 2022-02-17 | エフ・ホフマン-ラ・ロシュ・アクチェンゲゼルシャフト | Cd3に対する二重特異性抗体 |
TW201829463A (zh) | 2016-11-18 | 2018-08-16 | 瑞士商赫孚孟拉羅股份公司 | 抗hla-g抗體及其用途 |
CR20190297A (es) | 2016-11-23 | 2019-11-01 | Bioverativ Therapeutics Inc | Anticuerpos biespecíficos que se unen al factor de coagulación ix y al factor de coagulación x |
KR102317884B1 (ko) | 2016-12-21 | 2021-10-26 | 에프. 호프만-라 로슈 아게 | 항체의 시험관 내 당조작 |
BR112019009839A2 (pt) | 2016-12-21 | 2019-09-17 | Hoffmann La Roche | método para a produção enzimática de um anticorpo e anticorpo |
KR102293106B1 (ko) | 2016-12-21 | 2021-08-24 | 에프. 호프만-라 로슈 아게 | 항체의 시험관 내 당조작 방법 |
US20200291089A1 (en) | 2017-02-16 | 2020-09-17 | Elstar Therapeutics, Inc. | Multifunctional molecules comprising a trimeric ligand and uses thereof |
JP2020522254A (ja) | 2017-05-31 | 2020-07-30 | エルスター セラピューティクス, インコーポレイテッド | 骨髄増殖性白血病(mpl)タンパク質に結合する多特異性分子およびその使用 |
WO2019035938A1 (en) | 2017-08-16 | 2019-02-21 | Elstar Therapeutics, Inc. | MULTISPECIFIC MOLECULES BINDING TO BCMA AND USES THEREOF |
US20200300852A1 (en) | 2017-11-29 | 2020-09-24 | Hoffmann-La Roche Inc. | Target interference suppressed anti-drug antibody assay |
US11866498B2 (en) | 2018-02-08 | 2024-01-09 | Genentech, Inc. | Bispecific antigen-binding molecules and methods of use |
US20210009711A1 (en) | 2018-03-14 | 2021-01-14 | Elstar Therapeutics, Inc. | Multifunctional molecules and uses thereof |
US20210238280A1 (en) | 2018-03-14 | 2021-08-05 | Elstar Therapeutics, Inc. | Multifunctional molecules that bind to calreticulin and uses thereof |
AR114789A1 (es) | 2018-04-18 | 2020-10-14 | Hoffmann La Roche | Anticuerpos anti-hla-g y uso de los mismos |
AU2019265699A1 (en) | 2018-05-08 | 2021-01-07 | Amgen Inc. | Bispecific antibodies with cleavable C-terminal charge-paired tags |
US11845797B2 (en) | 2018-07-03 | 2023-12-19 | Marengo Therapeutics, Inc. | Anti-TCR antibody molecules and uses thereof |
CN112135626B (zh) | 2018-07-25 | 2023-05-23 | 信达生物制药(苏州)有限公司 | 抗tigit抗体及其用途 |
EP3608674A1 (en) | 2018-08-09 | 2020-02-12 | Regeneron Pharmaceuticals, Inc. | Methods for assessing binding affinity of an antibody variant to the neonatal fc receptor |
WO2020136566A1 (en) | 2018-12-24 | 2020-07-02 | Sanofi | Multispecific binding proteins with mutant fab domains |
WO2020136564A1 (en) * | 2018-12-24 | 2020-07-02 | Sanofi | Pseudofab-based multispecific binding proteins |
CN113227789A (zh) | 2018-12-30 | 2021-08-06 | 豪夫迈·罗氏有限公司 | 基于pH梯度SPR的结合测定 |
EP3927744A1 (en) | 2019-02-21 | 2021-12-29 | Marengo Therapeutics, Inc. | Multifunctional molecules that bind to t cell related cancer cells and uses thereof |
CN114127112A (zh) | 2019-02-21 | 2022-03-01 | 马伦戈治疗公司 | 与t细胞结合的多功能分子及其治疗自身免疫性病症的用途 |
GB2599229B (en) | 2019-02-21 | 2024-04-24 | Marengo Therapeutics Inc | Multifunctional molecules that bind to calreticulin and uses thereof |
SG11202108955QA (en) | 2019-02-21 | 2021-09-29 | Marengo Therapeutics Inc | Antibody molecules that bind to nkp30 and uses thereof |
AU2020226904A1 (en) | 2019-02-21 | 2021-09-16 | Marengo Therapeutics, Inc. | Anti-TCR antibody molecules and uses thereof |
EP3947440A1 (en) | 2019-03-29 | 2022-02-09 | F. Hoffmann-La Roche AG | Method for generating avid-binding multispecific antibodies |
WO2020200941A1 (en) | 2019-03-29 | 2020-10-08 | F. Hoffmann-La Roche Ag | Spr-based binding assay for the functional analysis of multivalent molecules |
CN113811770B (zh) | 2019-05-13 | 2024-06-28 | 豪夫迈·罗氏有限公司 | 抑制干扰的药代动力学免疫测定 |
MX2021015538A (es) | 2019-06-19 | 2022-02-10 | Hoffmann La Roche | Metodo para la generacion de una celula que expresa un anticuerpo biespecifico bivalente mediante la integracion dirigida de multiples casetes de expresion en una organizacion definida. |
CA3140192A1 (en) | 2019-06-19 | 2020-12-24 | Johannes Auer | Method for the generation of a multivalent, multispecific antibody expressing cell by targeted integration of multiple expression cassettes in a defined organization |
WO2020254352A1 (en) | 2019-06-19 | 2020-12-24 | F. Hoffmann-La Roche Ag | Method for the generation of a trivalent antibody expressing cell by targeted integration of multiple expression cassettes in a defined organization |
EP3986928A1 (en) | 2019-06-19 | 2022-04-27 | F. Hoffmann-La Roche AG | Method for the generation of a protein expressing cell by targeted integration using cre mrna |
CN113993887A (zh) | 2019-06-19 | 2022-01-28 | 豪夫迈·罗氏有限公司 | 通过以限定的组织形式靶向整合多个表达盒来产生多价双特异性抗体表达细胞的方法 |
EP3990646A1 (en) | 2019-06-26 | 2022-05-04 | F. Hoffmann-La Roche AG | Mammalian cell lines with sirt-1 gene knockout |
JP2022553640A (ja) | 2019-10-10 | 2022-12-26 | コディアック サイエンシーズ インコーポレイテッド | 眼障害を処置する方法 |
JP2023508596A (ja) | 2020-01-02 | 2023-03-02 | エフ. ホフマン-ラ ロシュ アーゲー | 脳内の治療用抗体の量を決定するための方法 |
EP4084821A4 (en) | 2020-01-03 | 2024-04-24 | Marengo Therapeutics, Inc. | CD33-BINDING MULTIFUNCTIONAL MOLECULES AND THEIR USES |
JP2023523011A (ja) | 2020-04-24 | 2023-06-01 | マレンゴ・セラピューティクス,インコーポレーテッド | T細胞関連のがん細胞に結合する多機能性分子およびその使用 |
WO2021254926A1 (en) | 2020-06-16 | 2021-12-23 | F. Hoffmann-La Roche Ag | Method for determining the free antigen of an antibody in a sample |
AU2021291011A1 (en) | 2020-06-19 | 2023-01-05 | F. Hoffmann-La Roche Ag | Antibodies binding to CD3 and CD19 |
AU2021331076A1 (en) | 2020-08-26 | 2023-04-06 | Marengo Therapeutics, Inc. | Antibody molecules that bind to NKp30 and uses thereof |
AU2021333779A1 (en) | 2020-08-26 | 2023-04-13 | Marengo Therapeutics, Inc. | Methods of detecting TRBC1 or TRBC2 |
JP2023542080A (ja) | 2020-08-26 | 2023-10-05 | マレンゴ・セラピューティクス,インコーポレーテッド | カルレティキュリンに結合する多機能性分子およびその使用 |
CA3191328A1 (en) | 2020-09-21 | 2022-03-24 | Genentech, Inc. | Purification of multispecific antibodies |
US20220154207A1 (en) | 2020-09-24 | 2022-05-19 | Hoffmann-La Roche Inc. | Mammalian cell lines with gene knockout |
KR20220082775A (ko) | 2020-12-10 | 2022-06-17 | 주식회사 유틸렉스 | 항-pd-1 항체 및 이의 용도 |
JP2024501662A (ja) | 2020-12-22 | 2024-01-15 | エフ. ホフマン-ラ ロシュ アーゲー | Xbp1を標的とするオリゴヌクレオチド |
CA3203257A1 (en) | 2020-12-23 | 2022-06-30 | Li Li | Anti-b7-h3 antibody and uses thereof |
CN114716548A (zh) | 2021-01-05 | 2022-07-08 | (株)爱恩德生物 | 抗-fgfr3抗体及其用途 |
EP4289865A1 (en) | 2021-02-04 | 2023-12-13 | Innovent Biologics (Suzhou) Co., Ltd. | Anti-tnfr2 antibody and use thereof |
JP2024512240A (ja) | 2021-02-18 | 2024-03-19 | エフ. ホフマン-ラ ロシュ アーゲー | 複雑な多段階の抗体相互作用を解明するための方法 |
BR112023020832A2 (pt) | 2021-04-08 | 2023-12-19 | Marengo Therapeutics Inc | Moléculas multifuncionais ligadas a tcr e seus usos |
JP2024513474A (ja) | 2021-04-09 | 2024-03-25 | エフ. ホフマン-ラ ロシュ アーゲー | 異種ポリペプチドを発現する細胞クローンを選択するための方法 |
JP2024517844A (ja) * | 2021-05-04 | 2024-04-23 | リジェネロン・ファーマシューティカルズ・インコーポレイテッド | 多重特異性fgf21受容体アゴニスト及びそれらの使用 |
CN117597365A (zh) * | 2021-05-04 | 2024-02-23 | 再生元制药公司 | 多特异性fgf21受体激动剂及其应用 |
CN113278071B (zh) | 2021-05-27 | 2021-12-21 | 江苏荃信生物医药股份有限公司 | 抗人干扰素α受体1单克隆抗体及其应用 |
CN113683694B (zh) | 2021-09-03 | 2022-05-13 | 江苏荃信生物医药股份有限公司 | 一种抗人tslp单克隆抗体及其应用 |
CN113603775B (zh) | 2021-09-03 | 2022-05-20 | 江苏荃信生物医药股份有限公司 | 抗人白介素-33单克隆抗体及其应用 |
CN118284809A (zh) | 2021-11-25 | 2024-07-02 | 豪夫迈·罗氏有限公司 | 少量抗体副产物的定量 |
WO2023117325A1 (en) | 2021-12-21 | 2023-06-29 | F. Hoffmann-La Roche Ag | Method for the determination of hydrolytic activity |
WO2023129974A1 (en) | 2021-12-29 | 2023-07-06 | Bristol-Myers Squibb Company | Generation of landing pad cell lines |
WO2023202967A1 (en) | 2022-04-19 | 2023-10-26 | F. Hoffmann-La Roche Ag | Improved production cells |
WO2023232961A1 (en) | 2022-06-03 | 2023-12-07 | F. Hoffmann-La Roche Ag | Improved production cells |
WO2024079069A1 (en) | 2022-10-12 | 2024-04-18 | F. Hoffmann-La Roche Ag | Method for classifying cells |
WO2024110426A1 (en) | 2022-11-23 | 2024-05-30 | F. Hoffmann-La Roche Ag | Method for increasing recombinant protein expression |
Family Cites Families (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
DE69922159T2 (de) | 1998-01-23 | 2005-12-01 | Vlaams Interuniversitair Instituut Voor Biotechnologie | Mehrzweck-antikörperderivate |
AU760854B2 (en) * | 1998-06-22 | 2003-05-22 | Immunomedics Inc. | Use of bi-specific antibodies for pre-targeting diagnosis and therapy |
AU2001247616B2 (en) | 2000-04-11 | 2007-06-14 | Genentech, Inc. | Multivalent antibodies and uses therefor |
RU2430927C2 (ru) * | 2000-10-20 | 2011-10-10 | Тугаи Сейяку Кабусики Кайся | Агонистическое соединение, способное специфически узнавать и поперечно сшивать молекулу клеточной поверхности или внутриклеточную молекулу |
EP1541319B1 (en) * | 2002-06-25 | 2011-10-05 | Sumitomo Bakelite Co., Ltd. | Device and method for processing carrier tape |
JO3000B1 (ar) | 2004-10-20 | 2016-09-05 | Genentech Inc | مركبات أجسام مضادة . |
JP2008531049A (ja) | 2005-02-28 | 2008-08-14 | セントカー・インコーポレーテツド | ヘテロ二量体タンパク質結合組成物 |
EP3623473A1 (en) | 2005-03-31 | 2020-03-18 | Chugai Seiyaku Kabushiki Kaisha | Process for production of polypeptide by regulation of assembly |
WO2007044887A2 (en) | 2005-10-11 | 2007-04-19 | Transtarget, Inc. | Method for producing a population of homogenous tetravalent bispecific antibodies |
FR2894959B1 (fr) | 2005-12-15 | 2008-02-29 | Galderma Res & Dev | Derives biphenyliques agonistes selectifs du recepteur rar-gamma |
US20070274985A1 (en) | 2006-05-26 | 2007-11-29 | Stefan Dubel | Antibody |
US8227577B2 (en) | 2007-12-21 | 2012-07-24 | Hoffman-La Roche Inc. | Bivalent, bispecific antibodies |
US20090162359A1 (en) | 2007-12-21 | 2009-06-25 | Christian Klein | Bivalent, bispecific antibodies |
US8242247B2 (en) | 2007-12-21 | 2012-08-14 | Hoffmann-La Roche Inc. | Bivalent, bispecific antibodies |
US9266967B2 (en) * | 2007-12-21 | 2016-02-23 | Hoffmann-La Roche, Inc. | Bivalent, bispecific antibodies |
MX2011010159A (es) | 2009-04-02 | 2011-10-17 | Roche Glycart Ag | Anticuerpos multiespecificos que comprenden anticuerpos de longitud completa y fragmentos fab de cadena sencilla. |
JP5616428B2 (ja) | 2009-04-07 | 2014-10-29 | ロシュ グリクアート アクチェンゲゼルシャフト | 三価の二重特異性抗体 |
EP2435473B1 (en) | 2009-05-27 | 2013-10-02 | F.Hoffmann-La Roche Ag | Tri- or tetraspecific antibodies |
US9676845B2 (en) | 2009-06-16 | 2017-06-13 | Hoffmann-La Roche, Inc. | Bispecific antigen binding proteins |
US8703132B2 (en) | 2009-06-18 | 2014-04-22 | Hoffmann-La Roche, Inc. | Bispecific, tetravalent antigen binding proteins |
AR085403A1 (es) | 2011-02-28 | 2013-09-25 | Hoffmann La Roche | Proteinas monovalentes que se unen a antigenos |
WO2012116926A1 (en) | 2011-02-28 | 2012-09-07 | F. Hoffmann-La Roche Ag | Antigen binding proteins |
PL2748202T3 (pl) | 2011-08-23 | 2018-12-31 | Roche Glycart Ag | Dwuswoiste cząsteczki wiążące antygen |
CA2837975C (en) | 2011-08-23 | 2022-04-05 | Roche Glycart Ag | Bispecific t cell activating antigen binding molecules |
-
2010
- 2010-05-25 US US12/786,477 patent/US8703132B2/en active Active
- 2010-06-14 BR BRPI1010109A patent/BRPI1010109A2/pt not_active IP Right Cessation
- 2010-06-14 CN CN201080036158.4A patent/CN102803295B/zh active Active
- 2010-06-14 RU RU2012101485/10A patent/RU2604189C2/ru not_active IP Right Cessation
- 2010-06-14 ES ES10724717.3T patent/ES2442917T3/es active Active
- 2010-06-14 AU AU2010262130A patent/AU2010262130A1/en not_active Abandoned
- 2010-06-14 PE PE2011002112A patent/PE20120414A1/es not_active Application Discontinuation
- 2010-06-14 MX MX2011013616A patent/MX2011013616A/es active IP Right Grant
- 2010-06-14 SG SG2011093655A patent/SG177272A1/en unknown
- 2010-06-14 WO PCT/EP2010/003560 patent/WO2010145793A1/en active Application Filing
- 2010-06-14 KR KR1020127001020A patent/KR101431344B1/ko not_active IP Right Cessation
- 2010-06-14 JP JP2012515382A patent/JP5689118B2/ja active Active
- 2010-06-14 CA CA2764393A patent/CA2764393C/en not_active Expired - Fee Related
- 2010-06-14 EP EP10724717.3A patent/EP2443153B1/en active Active
- 2010-06-16 AR ARP100102127A patent/AR077111A1/es unknown
- 2010-06-17 TW TW099119745A patent/TW201111394A/zh unknown
-
2011
- 2011-11-27 IL IL216620A patent/IL216620A0/en unknown
- 2011-12-05 ZA ZA2011/08922A patent/ZA201108922B/en unknown
- 2011-12-14 CL CL2011003149A patent/CL2011003149A1/es unknown
-
2013
- 2013-02-08 HK HK13101811.0A patent/HK1174645A1/zh unknown
-
2014
- 2014-04-18 US US14/256,891 patent/US20150044214A1/en not_active Abandoned
Also Published As
Publication number | Publication date |
---|---|
EP2443153A1 (en) | 2012-04-25 |
ZA201108922B (en) | 2012-08-29 |
RU2012101485A (ru) | 2013-07-27 |
PE20120414A1 (es) | 2012-05-04 |
KR20120028382A (ko) | 2012-03-22 |
US8703132B2 (en) | 2014-04-22 |
US20150044214A1 (en) | 2015-02-12 |
CA2764393C (en) | 2016-11-22 |
CA2764393A1 (en) | 2010-12-23 |
CN102803295A (zh) | 2012-11-28 |
HK1174645A1 (zh) | 2013-06-14 |
MX2011013616A (es) | 2012-01-19 |
US20100322934A1 (en) | 2010-12-23 |
EP2443153B1 (en) | 2013-11-06 |
ES2442917T3 (es) | 2014-02-14 |
RU2604189C2 (ru) | 2016-12-10 |
JP2012530089A (ja) | 2012-11-29 |
IL216620A0 (en) | 2012-02-29 |
AR077111A1 (es) | 2011-08-03 |
WO2010145793A1 (en) | 2010-12-23 |
KR101431344B1 (ko) | 2014-08-19 |
CL2011003149A1 (es) | 2012-07-20 |
AU2010262130A1 (en) | 2011-12-15 |
SG177272A1 (en) | 2012-02-28 |
CN102803295B (zh) | 2015-04-22 |
JP5689118B2 (ja) | 2015-03-25 |
BRPI1010109A2 (pt) | 2016-03-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
US11673945B2 (en) | Bispecific antigen binding proteins | |
US20210309730A1 (en) | Multispecific antibodies | |
JP5689118B2 (ja) | 二重特異性四価抗原結合タンパク質 | |
EP2435473B1 (en) | Tri- or tetraspecific antibodies | |
KR20150013188A (ko) | 다중특이적 항체 | |
TW201039850A (en) | Trivalent, bispecific antibodies | |
WO2013164325A1 (en) | Multispecific antigen binding proteins |