CN113372446A - 用于结合白细胞介素4受体的抗体 - Google Patents
用于结合白细胞介素4受体的抗体 Download PDFInfo
- Publication number
- CN113372446A CN113372446A CN202110760590.8A CN202110760590A CN113372446A CN 113372446 A CN113372446 A CN 113372446A CN 202110760590 A CN202110760590 A CN 202110760590A CN 113372446 A CN113372446 A CN 113372446A
- Authority
- CN
- China
- Prior art keywords
- seq
- variable region
- chain variable
- gly
- ser
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
- 230000027455 binding Effects 0.000 title claims abstract description 17
- 102000010787 Interleukin-4 Receptors Human genes 0.000 title description 4
- 108010038486 Interleukin-4 Receptors Proteins 0.000 title description 4
- 102000004388 Interleukin-4 Human genes 0.000 claims abstract description 33
- 108090000978 Interleukin-4 Proteins 0.000 claims abstract description 33
- 229940028885 interleukin-4 Drugs 0.000 claims abstract description 26
- 150000007523 nucleic acids Chemical group 0.000 claims abstract description 25
- 239000013598 vector Substances 0.000 claims abstract description 24
- 238000004519 manufacturing process Methods 0.000 claims abstract description 4
- 238000000034 method Methods 0.000 claims description 29
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 22
- 108020001507 fusion proteins Proteins 0.000 claims description 15
- 102000037865 fusion proteins Human genes 0.000 claims description 15
- 208000026935 allergic disease Diseases 0.000 claims description 13
- 208000006673 asthma Diseases 0.000 claims description 13
- 239000008194 pharmaceutical composition Substances 0.000 claims description 13
- 239000003814 drug Substances 0.000 claims description 11
- 206010039085 Rhinitis allergic Diseases 0.000 claims description 10
- 201000010105 allergic rhinitis Diseases 0.000 claims description 10
- 201000008937 atopic dermatitis Diseases 0.000 claims description 10
- 208000010668 atopic eczema Diseases 0.000 claims description 9
- 229940079593 drug Drugs 0.000 claims description 8
- ZFXYFBGIUFBOJW-UHFFFAOYSA-N theophylline Chemical compound O=C1N(C)C(=O)N(C)C2=C1NC=N2 ZFXYFBGIUFBOJW-UHFFFAOYSA-N 0.000 claims description 8
- 206010064212 Eosinophilic oesophagitis Diseases 0.000 claims description 7
- 201000000708 eosinophilic esophagitis Diseases 0.000 claims description 7
- 208000023275 Autoimmune disease Diseases 0.000 claims description 6
- 206010061218 Inflammation Diseases 0.000 claims description 6
- 230000002757 inflammatory effect Effects 0.000 claims description 6
- 230000004054 inflammatory process Effects 0.000 claims description 6
- 102000005962 receptors Human genes 0.000 claims description 6
- 108020003175 receptors Proteins 0.000 claims description 6
- 201000004624 Dermatitis Diseases 0.000 claims description 5
- 206010012434 Dermatitis allergic Diseases 0.000 claims description 5
- 239000000739 antihistaminic agent Substances 0.000 claims description 5
- 208000027866 inflammatory disease Diseases 0.000 claims description 5
- VOVIALXJUBGFJZ-KWVAZRHASA-N Budesonide Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@@H]2[C@@H]1[C@@H]1C[C@H]3OC(CCC)O[C@@]3(C(=O)CO)[C@@]1(C)C[C@@H]2O VOVIALXJUBGFJZ-KWVAZRHASA-N 0.000 claims description 4
- UCHDWCPVSPXUMX-TZIWLTJVSA-N Montelukast Chemical compound CC(C)(O)C1=CC=CC=C1CC[C@H](C=1C=C(\C=C\C=2N=C3C=C(Cl)C=CC3=CC=2)C=CC=1)SCC1(CC(O)=O)CC1 UCHDWCPVSPXUMX-TZIWLTJVSA-N 0.000 claims description 4
- 208000000592 Nasal Polyps Diseases 0.000 claims description 4
- QJJXYPPXXYFBGM-LFZNUXCKSA-N Tacrolimus Chemical compound C1C[C@@H](O)[C@H](OC)C[C@@H]1\C=C(/C)[C@@H]1[C@H](C)[C@@H](O)CC(=O)[C@H](CC=C)/C=C(C)/C[C@H](C)C[C@H](OC)[C@H]([C@H](C[C@H]2C)OC)O[C@@]2(O)C(=O)C(=O)N2CCCC[C@H]2C(=O)O1 QJJXYPPXXYFBGM-LFZNUXCKSA-N 0.000 claims description 4
- NDAUXUAQIAJITI-UHFFFAOYSA-N albuterol Chemical compound CC(C)(C)NCC(O)C1=CC=C(O)C(CO)=C1 NDAUXUAQIAJITI-UHFFFAOYSA-N 0.000 claims description 4
- 239000000924 antiasthmatic agent Substances 0.000 claims description 4
- NBMKJKDGKREAPL-DVTGEIKXSA-N beclomethasone Chemical compound C1CC2=CC(=O)C=C[C@]2(C)[C@]2(Cl)[C@@H]1[C@@H]1C[C@H](C)[C@@](C(=O)CO)(O)[C@@]1(C)C[C@@H]2O NBMKJKDGKREAPL-DVTGEIKXSA-N 0.000 claims description 4
- 229940092705 beclomethasone Drugs 0.000 claims description 4
- 229960004436 budesonide Drugs 0.000 claims description 4
- 229940082638 cardiac stimulant phosphodiesterase inhibitors Drugs 0.000 claims description 4
- 239000005556 hormone Substances 0.000 claims description 4
- 229940088597 hormone Drugs 0.000 claims description 4
- 239000003018 immunosuppressive agent Substances 0.000 claims description 4
- 229940125721 immunosuppressive agent Drugs 0.000 claims description 4
- 229960001361 ipratropium bromide Drugs 0.000 claims description 4
- KEWHKYJURDBRMN-ZEODDXGYSA-M ipratropium bromide hydrate Chemical compound O.[Br-].O([C@H]1C[C@H]2CC[C@@H](C1)[N@@+]2(C)C(C)C)C(=O)C(CO)C1=CC=CC=C1 KEWHKYJURDBRMN-ZEODDXGYSA-M 0.000 claims description 4
- 229960003088 loratadine Drugs 0.000 claims description 4
- JCCNYMKQOSZNPW-UHFFFAOYSA-N loratadine Chemical compound C1CN(C(=O)OCC)CCC1=C1C2=NC=CC=C2CCC2=CC(Cl)=CC=C21 JCCNYMKQOSZNPW-UHFFFAOYSA-N 0.000 claims description 4
- KBOPZPXVLCULAV-UHFFFAOYSA-N mesalamine Chemical compound NC1=CC=C(O)C(C(O)=O)=C1 KBOPZPXVLCULAV-UHFFFAOYSA-N 0.000 claims description 4
- 229960004963 mesalazine Drugs 0.000 claims description 4
- 229960005127 montelukast Drugs 0.000 claims description 4
- 208000016366 nasal cavity polyp Diseases 0.000 claims description 4
- 239000000041 non-steroidal anti-inflammatory agent Substances 0.000 claims description 4
- 229940021182 non-steroidal anti-inflammatory drug Drugs 0.000 claims description 4
- 239000000825 pharmaceutical preparation Substances 0.000 claims description 4
- 239000002571 phosphodiesterase inhibitor Substances 0.000 claims description 4
- KASDHRXLYQOAKZ-ZPSXYTITSA-N pimecrolimus Chemical compound C/C([C@H]1OC(=O)[C@@H]2CCCCN2C(=O)C(=O)[C@]2(O)O[C@@H]([C@H](C[C@H]2C)OC)[C@@H](OC)C[C@@H](C)C/C(C)=C/[C@H](C(C[C@H](O)[C@H]1C)=O)CC)=C\[C@@H]1CC[C@@H](Cl)[C@H](OC)C1 KASDHRXLYQOAKZ-ZPSXYTITSA-N 0.000 claims description 4
- 229960005330 pimecrolimus Drugs 0.000 claims description 4
- 206010039073 rheumatoid arthritis Diseases 0.000 claims description 4
- 229960002052 salbutamol Drugs 0.000 claims description 4
- 229960001967 tacrolimus Drugs 0.000 claims description 4
- QJJXYPPXXYFBGM-SHYZHZOCSA-N tacrolimus Natural products CO[C@H]1C[C@H](CC[C@@H]1O)C=C(C)[C@H]2OC(=O)[C@H]3CCCCN3C(=O)C(=O)[C@@]4(O)O[C@@H]([C@H](C[C@H]4C)OC)[C@@H](C[C@H](C)CC(=C[C@@H](CC=C)C(=O)C[C@H](O)[C@H]2C)C)OC QJJXYPPXXYFBGM-SHYZHZOCSA-N 0.000 claims description 4
- 229960000278 theophylline Drugs 0.000 claims description 4
- 108060003951 Immunoglobulin Proteins 0.000 claims description 3
- 102000018358 immunoglobulin Human genes 0.000 claims description 3
- 238000002347 injection Methods 0.000 claims description 3
- 239000007924 injection Substances 0.000 claims description 3
- 230000002265 prevention Effects 0.000 claims description 3
- 101001002709 Homo sapiens Interleukin-4 Proteins 0.000 claims description 2
- 241000124008 Mammalia Species 0.000 claims description 2
- 239000003937 drug carrier Substances 0.000 claims description 2
- 102000055229 human IL4 Human genes 0.000 claims description 2
- 239000003199 leukotriene receptor blocking agent Substances 0.000 claims description 2
- 102000003835 leukotriene receptors Human genes 0.000 claims description 2
- 108090000146 leukotriene receptors Proteins 0.000 claims description 2
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 2
- 239000003087 receptor blocking agent Substances 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 2
- 238000012258 culturing Methods 0.000 claims 1
- 210000004027 cell Anatomy 0.000 description 94
- 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 81
- PZTZYZUTCPZWJH-FXQIFTODSA-N Val-Ser-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PZTZYZUTCPZWJH-FXQIFTODSA-N 0.000 description 47
- YQPFCZVKMUVZIN-AUTRQRHGSA-N Glu-Val-Gln Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O YQPFCZVKMUVZIN-AUTRQRHGSA-N 0.000 description 46
- 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 46
- WNZOCXUOGVYYBJ-CDMKHQONSA-N Gly-Phe-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)NC(=O)CN)O WNZOCXUOGVYYBJ-CDMKHQONSA-N 0.000 description 46
- 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 46
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 46
- 108010067216 glycyl-glycyl-glycine Proteins 0.000 description 46
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 46
- 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 44
- ZDILXFDENZVOTL-BPNCWPANSA-N Ala-Val-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZDILXFDENZVOTL-BPNCWPANSA-N 0.000 description 42
- 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 42
- 108010089804 glycyl-threonine Proteins 0.000 description 41
- OYTPNWYZORARHL-XHNCKOQMSA-N Gln-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)N)N OYTPNWYZORARHL-XHNCKOQMSA-N 0.000 description 37
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 37
- FEHQLKKBVJHSEC-SZMVWBNQSA-N Leu-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC(C)C)C(O)=O)=CNC2=C1 FEHQLKKBVJHSEC-SZMVWBNQSA-N 0.000 description 37
- 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 37
- 108010008355 arginyl-glutamine Proteins 0.000 description 37
- 108010077037 tyrosyl-tyrosyl-phenylalanine Proteins 0.000 description 37
- IOVHBRCQOGWAQH-ZKWXMUAHSA-N Ser-Gly-Ile Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOVHBRCQOGWAQH-ZKWXMUAHSA-N 0.000 description 36
- AJBVYEYZVYPFCF-CIUDSAMLSA-N Ala-Lys-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(O)=O AJBVYEYZVYPFCF-CIUDSAMLSA-N 0.000 description 35
- HQIZDMIGUJOSNI-IUCAKERBSA-N Arg-Gly-Arg Chemical compound N[C@@H](CCCNC(N)=N)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(O)=O HQIZDMIGUJOSNI-IUCAKERBSA-N 0.000 description 35
- UVTGNSWSRSCPLP-UHFFFAOYSA-N Arg-Tyr Natural products NC(CCNC(=N)N)C(=O)NC(Cc1ccc(O)cc1)C(=O)O UVTGNSWSRSCPLP-UHFFFAOYSA-N 0.000 description 35
- JSHWXQIZOCVWIA-ZKWXMUAHSA-N Asp-Ser-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JSHWXQIZOCVWIA-ZKWXMUAHSA-N 0.000 description 35
- MSHXWFKYXJTLEZ-CIUDSAMLSA-N Gln-Met-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N MSHXWFKYXJTLEZ-CIUDSAMLSA-N 0.000 description 35
- QSDKBRMVXSWAQE-BFHQHQDPSA-N Gly-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)CN QSDKBRMVXSWAQE-BFHQHQDPSA-N 0.000 description 35
- KRRMJKMGWWXWDW-STQMWFEESA-N Gly-Arg-Phe Chemical compound NC(=N)NCCC[C@H](NC(=O)CN)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 KRRMJKMGWWXWDW-STQMWFEESA-N 0.000 description 35
- BYYNJRSNDARRBX-YFKPBYRVSA-N Gly-Gln-Gly Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O BYYNJRSNDARRBX-YFKPBYRVSA-N 0.000 description 35
- PDUHNKAFQXQNLH-ZETCQYMHSA-N Gly-Lys-Gly Chemical compound NCCCC[C@H](NC(=O)CN)C(=O)NCC(O)=O PDUHNKAFQXQNLH-ZETCQYMHSA-N 0.000 description 35
- 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 35
- JWOBLHJRDADHLN-KKUMJFAQSA-N Ser-Leu-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JWOBLHJRDADHLN-KKUMJFAQSA-N 0.000 description 35
- FQPDRTDDEZXCEC-SVSWQMSJSA-N Thr-Ile-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O FQPDRTDDEZXCEC-SVSWQMSJSA-N 0.000 description 35
- KZSYAEWQMJEGRZ-RHYQMDGZSA-N Thr-Leu-Val Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O KZSYAEWQMJEGRZ-RHYQMDGZSA-N 0.000 description 35
- TYGHOWWWMTWVKM-HJOGWXRNSA-N Tyr-Tyr-Phe 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=CC=1)C(O)=O)C1=CC=C(O)C=C1 TYGHOWWWMTWVKM-HJOGWXRNSA-N 0.000 description 35
- 239000000243 solution Substances 0.000 description 35
- 108010073969 valyllysine Proteins 0.000 description 35
- KXEVYGKATAMXJJ-ACZMJKKPSA-N Ala-Glu-Asp Chemical compound C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O KXEVYGKATAMXJJ-ACZMJKKPSA-N 0.000 description 34
- DGLQWAFPIXDKRL-UBHSHLNASA-N Ala-Met-Phe Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N DGLQWAFPIXDKRL-UBHSHLNASA-N 0.000 description 34
- PQWTZSNVWSOFFK-FXQIFTODSA-N Arg-Asp-Asn Chemical compound C(C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)CN=C(N)N PQWTZSNVWSOFFK-FXQIFTODSA-N 0.000 description 34
- WEDZFLRYSIDIRX-IHRRRGAJSA-N Phe-Ser-Arg Chemical compound NC(=N)NCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=CC=C1 WEDZFLRYSIDIRX-IHRRRGAJSA-N 0.000 description 34
- 108010008685 alanyl-glutamyl-aspartic acid Proteins 0.000 description 34
- 108010051242 phenylalanylserine Proteins 0.000 description 31
- NBTGEURICRTMGL-WHFBIAKZSA-N Ala-Gly-Ser Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O NBTGEURICRTMGL-WHFBIAKZSA-N 0.000 description 30
- QCBYAHHNOHBXIH-UWVGGRQHSA-N His-Pro-Gly Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)C1=CN=CN1 QCBYAHHNOHBXIH-UWVGGRQHSA-N 0.000 description 30
- MVJRBCJCRYGCKV-GVXVVHGQSA-N Leu-Val-Gln Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O MVJRBCJCRYGCKV-GVXVVHGQSA-N 0.000 description 30
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 30
- 108010079364 N-glycylalanine Proteins 0.000 description 23
- 108010087924 alanylproline Proteins 0.000 description 23
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 23
- JPPLRQVZMZFOSX-UWJYBYFXSA-N Asn-Tyr-Ala Chemical compound NC(=O)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=C(O)C=C1 JPPLRQVZMZFOSX-UWJYBYFXSA-N 0.000 description 22
- XZLLTYBONVKGLO-SDDRHHMPSA-N Gln-Lys-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCCN)NC(=O)[C@H](CCC(=O)N)N)C(=O)O XZLLTYBONVKGLO-SDDRHHMPSA-N 0.000 description 22
- CQZDZKRHFWJXDF-WDSKDSINSA-N Gly-Gln-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CN CQZDZKRHFWJXDF-WDSKDSINSA-N 0.000 description 22
- VCYJKOLZYPYGJV-AVGNSLFASA-N Pro-Arg-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(O)=O VCYJKOLZYPYGJV-AVGNSLFASA-N 0.000 description 22
- PKZIWSHDJYIPRH-JBACZVJFSA-N Trp-Tyr-Gln Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O PKZIWSHDJYIPRH-JBACZVJFSA-N 0.000 description 22
- SMLCYZYQFRTLCO-UWJYBYFXSA-N Tyr-Cys-Ala Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O SMLCYZYQFRTLCO-UWJYBYFXSA-N 0.000 description 22
- GULIUBBXCYPDJU-CQDKDKBSSA-N Tyr-Leu-Ala Chemical compound [O-]C(=O)[C@H](C)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H]([NH3+])CC1=CC=C(O)C=C1 GULIUBBXCYPDJU-CQDKDKBSSA-N 0.000 description 22
- 108010035534 tyrosyl-leucyl-alanine Proteins 0.000 description 22
- MRQQMVZUHXUPEV-IHRRRGAJSA-N Asp-Arg-Phe Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O MRQQMVZUHXUPEV-IHRRRGAJSA-N 0.000 description 21
- AVZHGSCDKIQZPQ-CIUDSAMLSA-N Glu-Arg-Ala Chemical compound C[C@H](NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@@H](N)CCC(O)=O)C(O)=O AVZHGSCDKIQZPQ-CIUDSAMLSA-N 0.000 description 21
- INGJLBQKTRJLFO-UKJIMTQDSA-N Glu-Ile-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCC(O)=O INGJLBQKTRJLFO-UKJIMTQDSA-N 0.000 description 21
- BHPQOIPBLYJNAW-NGZCFLSTSA-N Gly-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN BHPQOIPBLYJNAW-NGZCFLSTSA-N 0.000 description 21
- SOEGEPHNZOISMT-BYPYZUCNSA-N Gly-Ser-Gly Chemical compound NCC(=O)N[C@@H](CO)C(=O)NCC(O)=O SOEGEPHNZOISMT-BYPYZUCNSA-N 0.000 description 21
- 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 21
- SBANPBVRHYIMRR-GARJFASQSA-N Leu-Ser-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CO)C(=O)N1CCC[C@@H]1C(=O)O)N SBANPBVRHYIMRR-GARJFASQSA-N 0.000 description 21
- 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 21
- LCNASHSOFMRYFO-WDCWCFNPSA-N Leu-Thr-Gln 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)CCC(N)=O LCNASHSOFMRYFO-WDCWCFNPSA-N 0.000 description 21
- LJBVRCDPWOJOEK-PPCPHDFISA-N Leu-Thr-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O LJBVRCDPWOJOEK-PPCPHDFISA-N 0.000 description 21
- YYKZDTVQHTUKDW-RYUDHWBXSA-N Phe-Gly-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N YYKZDTVQHTUKDW-RYUDHWBXSA-N 0.000 description 21
- 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 21
- RHAPJNVNWDBFQI-BQBZGAKWSA-N Ser-Pro-Gly Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O RHAPJNVNWDBFQI-BQBZGAKWSA-N 0.000 description 21
- VCAWFLIWYNMHQP-UKJIMTQDSA-N Val-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](C(C)C)N VCAWFLIWYNMHQP-UKJIMTQDSA-N 0.000 description 21
- 108010017391 lysylvaline Proteins 0.000 description 21
- NCQMBSJGJMYKCK-ZLUOBGJFSA-N Ala-Ser-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O NCQMBSJGJMYKCK-ZLUOBGJFSA-N 0.000 description 20
- USNJAPJZSGTTPX-XVSYOHENSA-N Asp-Phe-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O USNJAPJZSGTTPX-XVSYOHENSA-N 0.000 description 20
- SZQCDCKIGWQAQN-FXQIFTODSA-N Cys-Arg-Ala Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O SZQCDCKIGWQAQN-FXQIFTODSA-N 0.000 description 20
- 241000880493 Leptailurus serval Species 0.000 description 20
- WUGMRIBZSVSJNP-UHFFFAOYSA-N N-L-alanyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C)C(O)=O)=CNC2=C1 WUGMRIBZSVSJNP-UHFFFAOYSA-N 0.000 description 20
- MGDFPGCFVJFITQ-CIUDSAMLSA-N Pro-Glu-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O MGDFPGCFVJFITQ-CIUDSAMLSA-N 0.000 description 20
- FMDHKPRACUXATF-ACZMJKKPSA-N Ser-Gln-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O FMDHKPRACUXATF-ACZMJKKPSA-N 0.000 description 20
- SFTZWNJFZYOLBD-ZDLURKLDSA-N Ser-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO SFTZWNJFZYOLBD-ZDLURKLDSA-N 0.000 description 20
- QOIKZODVIPOPDD-AVGNSLFASA-N Tyr-Cys-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(O)=O QOIKZODVIPOPDD-AVGNSLFASA-N 0.000 description 20
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 20
- HTSSXFASOUSJQG-IHPCNDPISA-N Asp-Tyr-Trp 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=CNC2=C1C=CC=C2)C(O)=O HTSSXFASOUSJQG-IHPCNDPISA-N 0.000 description 18
- 108010038745 tryptophylglycine Proteins 0.000 description 18
- 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 17
- MCPXQHVVCPTRIM-HJOGWXRNSA-N Pro-Trp-Trp Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)O)C(=O)[C@@H]1CCCN1 MCPXQHVVCPTRIM-HJOGWXRNSA-N 0.000 description 17
- FQCILXROGNOZON-YUMQZZPRSA-N Gln-Pro-Gly Chemical compound NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O FQCILXROGNOZON-YUMQZZPRSA-N 0.000 description 16
- 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 16
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 16
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 16
- 238000004113 cell culture Methods 0.000 description 14
- 239000002609 medium Substances 0.000 description 14
- PIQRHJQWEPWFJG-UWJYBYFXSA-N Ser-Tyr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O PIQRHJQWEPWFJG-UWJYBYFXSA-N 0.000 description 13
- 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 13
- 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 13
- 108010009962 valyltyrosine Proteins 0.000 description 13
- OTOXOKCIIQLMFH-KZVJFYERSA-N Arg-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N OTOXOKCIIQLMFH-KZVJFYERSA-N 0.000 description 12
- OACQOWPRWGNKTP-AVGNSLFASA-N Gln-Tyr-Asp Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(O)=O)C(O)=O OACQOWPRWGNKTP-AVGNSLFASA-N 0.000 description 12
- UAELWXJFLZBKQS-WHOFXGATSA-N Ile-Phe-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](Cc1ccccc1)C(=O)NCC(O)=O UAELWXJFLZBKQS-WHOFXGATSA-N 0.000 description 12
- 108010061238 threonyl-glycine Proteins 0.000 description 12
- 101100069857 Caenorhabditis elegans hil-4 gene Proteins 0.000 description 11
- VBZXFFYOBDLLFE-HSHDSVGOSA-N Pro-Trp-Thr Chemical compound N([C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H]([C@H](O)C)C(O)=O)C(=O)[C@@H]1CCCN1 VBZXFFYOBDLLFE-HSHDSVGOSA-N 0.000 description 11
- 239000012228 culture supernatant Substances 0.000 description 11
- 238000001514 detection method Methods 0.000 description 11
- 230000014509 gene expression Effects 0.000 description 11
- 239000000523 sample Substances 0.000 description 10
- UMBDRSMLCUYIRI-DVJZZOLTSA-N Gly-Trp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)CN)O UMBDRSMLCUYIRI-DVJZZOLTSA-N 0.000 description 9
- 210000002966 serum Anatomy 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- DBKNLHKEVPZVQC-LPEHRKFASA-N Arg-Ala-Pro Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@@H]1C(O)=O DBKNLHKEVPZVQC-LPEHRKFASA-N 0.000 description 8
- 238000002965 ELISA Methods 0.000 description 8
- SGVGIVDZLSHSEN-RYUDHWBXSA-N Gln-Tyr-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O SGVGIVDZLSHSEN-RYUDHWBXSA-N 0.000 description 8
- KAXZXLSXFWSNNZ-XVYDVKMFSA-N His-Ser-Ala Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O KAXZXLSXFWSNNZ-XVYDVKMFSA-N 0.000 description 8
- PRTZQMBYUZFSFA-XEGUGMAKSA-N Ile-Tyr-Gly Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)NCC(=O)O)N PRTZQMBYUZFSFA-XEGUGMAKSA-N 0.000 description 8
- KSFXWENSJABBFI-ZKWXMUAHSA-N Val-Ser-Asn Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KSFXWENSJABBFI-ZKWXMUAHSA-N 0.000 description 8
- 210000004369 blood Anatomy 0.000 description 8
- 239000008280 blood Substances 0.000 description 8
- 230000002401 inhibitory effect Effects 0.000 description 8
- 239000013642 negative control Substances 0.000 description 8
- CUXJENOFJXOSOZ-BIIVOSGPSA-N Ser-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CO)N)C(=O)O CUXJENOFJXOSOZ-BIIVOSGPSA-N 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 102000004127 Cytokines Human genes 0.000 description 6
- 108090000695 Cytokines Proteins 0.000 description 6
- 206010012438 Dermatitis atopic Diseases 0.000 description 6
- 241000282567 Macaca fascicularis Species 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 6
- 239000000427 antigen Substances 0.000 description 6
- 102000036639 antigens Human genes 0.000 description 6
- 108091007433 antigens Proteins 0.000 description 6
- 239000011248 coating agent Substances 0.000 description 6
- 238000000576 coating method Methods 0.000 description 6
- 239000013604 expression vector Substances 0.000 description 6
- 238000001943 fluorescence-activated cell sorting Methods 0.000 description 6
- 210000001616 monocyte Anatomy 0.000 description 6
- 239000013641 positive control Substances 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- 108090000790 Enzymes Proteins 0.000 description 5
- 102000004190 Enzymes Human genes 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 5
- 210000003719 b-lymphocyte Anatomy 0.000 description 5
- 230000000903 blocking effect Effects 0.000 description 5
- 230000004663 cell proliferation Effects 0.000 description 5
- 239000006285 cell suspension Substances 0.000 description 5
- 239000003153 chemical reaction reagent Substances 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- 108090000623 proteins and genes Proteins 0.000 description 5
- JUJBNYBVVQSIOU-UHFFFAOYSA-M sodium;4-[2-(4-iodophenyl)-3-(4-nitrophenyl)tetrazol-2-ium-5-yl]benzene-1,3-disulfonate Chemical compound [Na+].C1=CC([N+](=O)[O-])=CC=C1N1[N+](C=2C=CC(I)=CC=2)=NC(C=2C(=CC(=CC=2)S([O-])(=O)=O)S([O-])(=O)=O)=N1 JUJBNYBVVQSIOU-UHFFFAOYSA-M 0.000 description 5
- 230000003068 static effect Effects 0.000 description 5
- 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 4
- LOJYQMFIIJVETK-WDSKDSINSA-N Gln-Gln Chemical compound NC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(O)=O LOJYQMFIIJVETK-WDSKDSINSA-N 0.000 description 4
- 108010017213 Granulocyte-Macrophage Colony-Stimulating Factor Proteins 0.000 description 4
- 102100039620 Granulocyte-macrophage colony-stimulating factor Human genes 0.000 description 4
- GIRSNERMXCMDBO-GARJFASQSA-N His-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O GIRSNERMXCMDBO-GARJFASQSA-N 0.000 description 4
- LHSGPCFBGJHPCY-UHFFFAOYSA-N L-leucine-L-tyrosine Natural products CC(C)CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 LHSGPCFBGJHPCY-UHFFFAOYSA-N 0.000 description 4
- FGWUALWGCZJQDJ-URLPEUOOSA-N Phe-Thr-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O FGWUALWGCZJQDJ-URLPEUOOSA-N 0.000 description 4
- 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 4
- 210000001744 T-lymphocyte Anatomy 0.000 description 4
- NKUGCYDFQKFVOJ-JYJNAYRXSA-N Tyr-Leu-Gln Chemical compound NC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 NKUGCYDFQKFVOJ-JYJNAYRXSA-N 0.000 description 4
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 4
- 230000000694 effects Effects 0.000 description 4
- 108010012058 leucyltyrosine Proteins 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 230000008506 pathogenesis Effects 0.000 description 4
- 239000003755 preservative agent Substances 0.000 description 4
- 230000002335 preservative effect Effects 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 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 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 3
- 206010020751 Hypersensitivity Diseases 0.000 description 3
- 150000001413 amino acids Chemical group 0.000 description 3
- 238000003556 assay Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 210000003979 eosinophil Anatomy 0.000 description 3
- 210000003630 histaminocyte Anatomy 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 230000001404 mediated effect Effects 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 239000008188 pellet Substances 0.000 description 3
- 230000001737 promoting effect Effects 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- 230000009870 specific binding Effects 0.000 description 3
- 238000003860 storage Methods 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000001890 transfection Methods 0.000 description 3
- 230000010474 transient expression Effects 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- GIVATXIGCXFQQA-FXQIFTODSA-N Arg-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N GIVATXIGCXFQQA-FXQIFTODSA-N 0.000 description 2
- DCGLNNVKIZXQOJ-FXQIFTODSA-N Arg-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N DCGLNNVKIZXQOJ-FXQIFTODSA-N 0.000 description 2
- 241000283707 Capra Species 0.000 description 2
- 108010001857 Cell Surface Receptors Proteins 0.000 description 2
- 108091026890 Coding region Proteins 0.000 description 2
- 208000035473 Communicable disease Diseases 0.000 description 2
- OTQSTOXRUBVWAP-NRPADANISA-N Gln-Ser-Val Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O OTQSTOXRUBVWAP-NRPADANISA-N 0.000 description 2
- LJPIRKICOISLKN-WHFBIAKZSA-N Gly-Ala-Ser Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O LJPIRKICOISLKN-WHFBIAKZSA-N 0.000 description 2
- XUORRGAFUQIMLC-STQMWFEESA-N Gly-Arg-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)CN)O XUORRGAFUQIMLC-STQMWFEESA-N 0.000 description 2
- XTQFHTHIAKKCTM-YFKPBYRVSA-N Gly-Glu-Gly Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O XTQFHTHIAKKCTM-YFKPBYRVSA-N 0.000 description 2
- HMHRTKOWRUPPNU-RCOVLWMOSA-N Gly-Ile-Gly Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(O)=O HMHRTKOWRUPPNU-RCOVLWMOSA-N 0.000 description 2
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 2
- HGCNKOLVKRAVHD-UHFFFAOYSA-N L-Met-L-Phe Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 HGCNKOLVKRAVHD-UHFFFAOYSA-N 0.000 description 2
- LINKCQUOMUDLKN-KATARQTJSA-N Leu-Thr-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(C)C)N)O LINKCQUOMUDLKN-KATARQTJSA-N 0.000 description 2
- 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 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 229940127593 SEQ-9 Drugs 0.000 description 2
- BTKUIVBNGBFTTP-WHFBIAKZSA-N Ser-Ala-Gly Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)NCC(O)=O BTKUIVBNGBFTTP-WHFBIAKZSA-N 0.000 description 2
- GXXTUIUYTWGPMV-FXQIFTODSA-N Ser-Arg-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O GXXTUIUYTWGPMV-FXQIFTODSA-N 0.000 description 2
- DINQYZRMXGWWTG-GUBZILKMSA-N Ser-Pro-Pro Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DINQYZRMXGWWTG-GUBZILKMSA-N 0.000 description 2
- 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 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- XPNSAQMEAVSQRD-FBCQKBJTSA-N Thr-Gly-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)NCC(=O)NCC(O)=O XPNSAQMEAVSQRD-FBCQKBJTSA-N 0.000 description 2
- UIRPULWLRODAEQ-QEJZJMRPSA-N Trp-Ser-Glu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 UIRPULWLRODAEQ-QEJZJMRPSA-N 0.000 description 2
- BURPTJBFWIOHEY-UWJYBYFXSA-N Tyr-Ala-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 BURPTJBFWIOHEY-UWJYBYFXSA-N 0.000 description 2
- QNJYPWZACBACER-KKUMJFAQSA-N Tyr-Asp-His Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N)O QNJYPWZACBACER-KKUMJFAQSA-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
- 230000007815 allergy Effects 0.000 description 2
- 108010050025 alpha-glutamyltryptophan Proteins 0.000 description 2
- 230000004071 biological effect Effects 0.000 description 2
- 230000008827 biological function Effects 0.000 description 2
- 229960000074 biopharmaceutical Drugs 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 229950003468 dupilumab Drugs 0.000 description 2
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 238000001502 gel electrophoresis Methods 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 108010018006 histidylserine Proteins 0.000 description 2
- 230000002519 immonomodulatory effect Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 230000001788 irregular Effects 0.000 description 2
- 229960002725 isoflurane Drugs 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 102000006240 membrane receptors Human genes 0.000 description 2
- 108010068488 methionylphenylalanine Proteins 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 229920001184 polypeptide Polymers 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 102000004196 processed proteins & peptides Human genes 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 235000011149 sulphuric acid Nutrition 0.000 description 2
- 238000004114 suspension culture Methods 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 230000001052 transient effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- AXFMEGAFCUULFV-BLFANLJRSA-N (2s)-2-[[(2s)-1-[(2s,3r)-2-amino-3-methylpentanoyl]pyrrolidine-2-carbonyl]amino]pentanedioic acid Chemical compound CC[C@@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O AXFMEGAFCUULFV-BLFANLJRSA-N 0.000 description 1
- MPLOSMWGDNJSEV-WHFBIAKZSA-N Ala-Gly-Asp Chemical compound [H]N[C@@H](C)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O MPLOSMWGDNJSEV-WHFBIAKZSA-N 0.000 description 1
- LJFNNUBZSZCZFN-WHFBIAKZSA-N Ala-Gly-Cys Chemical compound N[C@@H](C)C(=O)NCC(=O)N[C@@H](CS)C(=O)O LJFNNUBZSZCZFN-WHFBIAKZSA-N 0.000 description 1
- MAEQBGQTDWDSJQ-LSJOCFKGSA-N Ala-Met-His Chemical compound C[C@@H](C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N MAEQBGQTDWDSJQ-LSJOCFKGSA-N 0.000 description 1
- XQNRANMFRPCFFW-GCJQMDKQSA-N Ala-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](C)N)O XQNRANMFRPCFFW-GCJQMDKQSA-N 0.000 description 1
- KTXKIYXZQFWJKB-VZFHVOOUSA-N Ala-Thr-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O KTXKIYXZQFWJKB-VZFHVOOUSA-N 0.000 description 1
- ZVWXMTTZJKBJCI-BHDSKKPTSA-N Ala-Trp-Ala Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](C)C(O)=O)=CNC2=C1 ZVWXMTTZJKBJCI-BHDSKKPTSA-N 0.000 description 1
- 206010049153 Allergic sinusitis Diseases 0.000 description 1
- OZNSCVPYWZRQPY-CIUDSAMLSA-N Arg-Asp-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O OZNSCVPYWZRQPY-CIUDSAMLSA-N 0.000 description 1
- ISVACHFCVRKIDG-SRVKXCTJSA-N Arg-Val-Arg Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O ISVACHFCVRKIDG-SRVKXCTJSA-N 0.000 description 1
- PCKRJVZAQZWNKM-WHFBIAKZSA-N Asn-Asn-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O PCKRJVZAQZWNKM-WHFBIAKZSA-N 0.000 description 1
- GJFYPBDMUGGLFR-NKWVEPMBSA-N Asn-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CC(=O)N)N)C(=O)O GJFYPBDMUGGLFR-NKWVEPMBSA-N 0.000 description 1
- IKLAUGBIDCDFOY-SRVKXCTJSA-N Asn-His-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(C)C)C(O)=O IKLAUGBIDCDFOY-SRVKXCTJSA-N 0.000 description 1
- UXHYOWXTJLBEPG-GSSVUCPTSA-N Asn-Thr-Thr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UXHYOWXTJLBEPG-GSSVUCPTSA-N 0.000 description 1
- GYWQGGUCMDCUJE-DLOVCJGASA-N Asp-Phe-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C)C(O)=O GYWQGGUCMDCUJE-DLOVCJGASA-N 0.000 description 1
- LIJXJYGRSRWLCJ-IHRRRGAJSA-N Asp-Phe-Arg Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O LIJXJYGRSRWLCJ-IHRRRGAJSA-N 0.000 description 1
- KPSHWSWFPUDEGF-FXQIFTODSA-N Asp-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC(O)=O KPSHWSWFPUDEGF-FXQIFTODSA-N 0.000 description 1
- PLNJUJGNLDSFOP-UWJYBYFXSA-N Asp-Tyr-Ala 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](C)C(O)=O PLNJUJGNLDSFOP-UWJYBYFXSA-N 0.000 description 1
- 208000012657 Atopic disease Diseases 0.000 description 1
- 102000019034 Chemokines Human genes 0.000 description 1
- 108010012236 Chemokines Proteins 0.000 description 1
- PZVJDMJHKUWSIV-AVGNSLFASA-N Gln-Cys-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CCC(=O)N)N)O PZVJDMJHKUWSIV-AVGNSLFASA-N 0.000 description 1
- KVXVVDFOZNYYKZ-DCAQKATOSA-N Gln-Gln-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O KVXVVDFOZNYYKZ-DCAQKATOSA-N 0.000 description 1
- VOLVNCMGXWDDQY-LPEHRKFASA-N Gln-Glu-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CCC(=O)N)N)C(=O)O VOLVNCMGXWDDQY-LPEHRKFASA-N 0.000 description 1
- JJKKWYQVHRUSDG-GUBZILKMSA-N Glu-Ala-Lys Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O JJKKWYQVHRUSDG-GUBZILKMSA-N 0.000 description 1
- DXVOKNVIKORTHQ-GUBZILKMSA-N Glu-Pro-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O DXVOKNVIKORTHQ-GUBZILKMSA-N 0.000 description 1
- FMVLWTYYODVFRG-BQBZGAKWSA-N Gly-Asn-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)CN FMVLWTYYODVFRG-BQBZGAKWSA-N 0.000 description 1
- JSLVAHYTAJJEQH-QWRGUYRKSA-N Gly-Ser-Phe Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JSLVAHYTAJJEQH-QWRGUYRKSA-N 0.000 description 1
- 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 1
- 102100029100 Hematopoietic prostaglandin D synthase Human genes 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- HRGGKHFHRSFSDE-CIUDSAMLSA-N His-Asn-Ser Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CO)C(=O)O)N HRGGKHFHRSFSDE-CIUDSAMLSA-N 0.000 description 1
- FBVHRDXSCYELMI-PBCZWWQYSA-N His-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N)O FBVHRDXSCYELMI-PBCZWWQYSA-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
- PUFNQIPSRXVLQJ-IHRRRGAJSA-N His-Val-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N PUFNQIPSRXVLQJ-IHRRRGAJSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 101000988802 Homo sapiens Hematopoietic prostaglandin D synthase Proteins 0.000 description 1
- 101001033312 Homo sapiens Interleukin-4 receptor subunit alpha Proteins 0.000 description 1
- 101000878605 Homo sapiens Low affinity immunoglobulin epsilon Fc receptor Proteins 0.000 description 1
- FXJLRZFMKGHYJP-CFMVVWHZSA-N Ile-Tyr-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N FXJLRZFMKGHYJP-CFMVVWHZSA-N 0.000 description 1
- 102100020793 Interleukin-13 receptor subunit alpha-2 Human genes 0.000 description 1
- 108010002616 Interleukin-5 Proteins 0.000 description 1
- FADYJNXDPBKVCA-UHFFFAOYSA-N L-Phenylalanyl-L-lysin Natural products NCCCCC(C(O)=O)NC(=O)C(N)CC1=CC=CC=C1 FADYJNXDPBKVCA-UHFFFAOYSA-N 0.000 description 1
- RCFDOSNHHZGBOY-UHFFFAOYSA-N L-isoleucyl-L-alanine Natural products CCC(C)C(N)C(=O)NC(C)C(O)=O RCFDOSNHHZGBOY-UHFFFAOYSA-N 0.000 description 1
- UILIPCLTHRPCRB-XUXIUFHCSA-N Leu-Arg-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(C)C)N UILIPCLTHRPCRB-XUXIUFHCSA-N 0.000 description 1
- OGUUKPXUTHOIAV-SDDRHHMPSA-N Leu-Glu-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N1CCC[C@@H]1C(=O)O)N OGUUKPXUTHOIAV-SDDRHHMPSA-N 0.000 description 1
- QLDHBYRUNQZIJQ-DKIMLUQUSA-N Leu-Ile-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O QLDHBYRUNQZIJQ-DKIMLUQUSA-N 0.000 description 1
- NRFGTHFONZYFNY-MGHWNKPDSA-N Leu-Ile-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 NRFGTHFONZYFNY-MGHWNKPDSA-N 0.000 description 1
- IEWBEPKLKUXQBU-VOAKCMCISA-N Leu-Leu-Thr Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O IEWBEPKLKUXQBU-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
- ZGGVHTQAPHVMKM-IHPCNDPISA-N Leu-Trp-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CCCCN)C(=O)O)N ZGGVHTQAPHVMKM-IHPCNDPISA-N 0.000 description 1
- 102100038007 Low affinity immunoglobulin epsilon Fc receptor Human genes 0.000 description 1
- YTJFXEDRUOQGSP-DCAQKATOSA-N Lys-Pro-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O YTJFXEDRUOQGSP-DCAQKATOSA-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
- 239000012515 MabSelect SuRe Substances 0.000 description 1
- 239000005984 Mepiquat Substances 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- ZENDEDYRYVHBEG-SRVKXCTJSA-N Phe-Asp-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 ZENDEDYRYVHBEG-SRVKXCTJSA-N 0.000 description 1
- NHDVNAKDACFHPX-GUBZILKMSA-N Pro-Arg-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O NHDVNAKDACFHPX-GUBZILKMSA-N 0.000 description 1
- CJZTUKSFZUSNCC-FXQIFTODSA-N Pro-Asp-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H]1CCCN1 CJZTUKSFZUSNCC-FXQIFTODSA-N 0.000 description 1
- UUHXBJHVTVGSKM-BQBZGAKWSA-N Pro-Gly-Asn Chemical compound [H]N1CCC[C@H]1C(=O)NCC(=O)N[C@@H](CC(N)=O)C(O)=O UUHXBJHVTVGSKM-BQBZGAKWSA-N 0.000 description 1
- JIWJRKNYLSHONY-KKUMJFAQSA-N Pro-Phe-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JIWJRKNYLSHONY-KKUMJFAQSA-N 0.000 description 1
- IALSFJSONJZBKB-HRCADAONSA-N Pro-Tyr-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N3CCC[C@@H]3C(=O)O IALSFJSONJZBKB-HRCADAONSA-N 0.000 description 1
- RDFQNDHEHVSONI-ZLUOBGJFSA-N Ser-Asn-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O RDFQNDHEHVSONI-ZLUOBGJFSA-N 0.000 description 1
- BTPAWKABYQMKKN-LKXGYXEUSA-N Ser-Asp-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BTPAWKABYQMKKN-LKXGYXEUSA-N 0.000 description 1
- HEQPKICPPDOSIN-SRVKXCTJSA-N Ser-Asp-Tyr Chemical compound OC[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 HEQPKICPPDOSIN-SRVKXCTJSA-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
- UIPXCLNLUUAMJU-JBDRJPRFSA-N Ser-Ile-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O UIPXCLNLUUAMJU-JBDRJPRFSA-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
- 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 1
- 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 1
- FGBLCMLXHRPVOF-IHRRRGAJSA-N Ser-Tyr-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FGBLCMLXHRPVOF-IHRRRGAJSA-N 0.000 description 1
- 230000006052 T cell proliferation Effects 0.000 description 1
- 210000004241 Th2 cell Anatomy 0.000 description 1
- VASYSJHSMSBTDU-LKXGYXEUSA-N Thr-Asn-Cys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CS)C(=O)O)N)O VASYSJHSMSBTDU-LKXGYXEUSA-N 0.000 description 1
- KRPKYGOFYUNIGM-XVSYOHENSA-N Thr-Asp-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N)O KRPKYGOFYUNIGM-XVSYOHENSA-N 0.000 description 1
- DHPPWTOLRWYIDS-XKBZYTNZSA-N Thr-Cys-Glu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(O)=O)C(O)=O DHPPWTOLRWYIDS-XKBZYTNZSA-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
- MECLEFZMPPOEAC-VOAKCMCISA-N Thr-Leu-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N)O MECLEFZMPPOEAC-VOAKCMCISA-N 0.000 description 1
- QFCQNHITJPRQTB-IEGACIPQSA-N Thr-Lys-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N)O QFCQNHITJPRQTB-IEGACIPQSA-N 0.000 description 1
- LXXCHJKHJYRMIY-FQPOAREZSA-N Thr-Tyr-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](C)C(O)=O LXXCHJKHJYRMIY-FQPOAREZSA-N 0.000 description 1
- CXUFDWZBHKUGKK-CABZTGNLSA-N Trp-Ala-Gly Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O)=CNC2=C1 CXUFDWZBHKUGKK-CABZTGNLSA-N 0.000 description 1
- SVGAWGVHFIYAEE-JSGCOSHPSA-N Trp-Gly-Gln Chemical compound C1=CC=C2C(C[C@H](N)C(=O)NCC(=O)N[C@@H](CCC(N)=O)C(O)=O)=CNC2=C1 SVGAWGVHFIYAEE-JSGCOSHPSA-N 0.000 description 1
- FXHOCONKLLUOCF-WDSOQIARSA-N Trp-Lys-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N FXHOCONKLLUOCF-WDSOQIARSA-N 0.000 description 1
- KBKTUNYBNJWFRL-UBHSHLNASA-N Trp-Ser-Asn Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O)=CNC2=C1 KBKTUNYBNJWFRL-UBHSHLNASA-N 0.000 description 1
- HTHCZRWCFXMENJ-KKUMJFAQSA-N Tyr-Arg-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O HTHCZRWCFXMENJ-KKUMJFAQSA-N 0.000 description 1
- OLYXUGBVBGSZDN-ACRUOGEOSA-N Tyr-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=C(O)C=C1 OLYXUGBVBGSZDN-ACRUOGEOSA-N 0.000 description 1
- 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 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
- LIQJSDDOULTANC-QSFUFRPTSA-N Val-Asn-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](C(C)C)N LIQJSDDOULTANC-QSFUFRPTSA-N 0.000 description 1
- YLHLNFUXDBOAGX-DCAQKATOSA-N Val-Cys-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N YLHLNFUXDBOAGX-DCAQKATOSA-N 0.000 description 1
- YLBNZCJFSVJDRJ-KJEVXHAQSA-N Val-Thr-Tyr Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](Cc1ccc(O)cc1)C(O)=O YLBNZCJFSVJDRJ-KJEVXHAQSA-N 0.000 description 1
- 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 1
- 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 1
- 108010081404 acein-2 Proteins 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 108010078114 alanyl-tryptophyl-alanine Proteins 0.000 description 1
- 108010041407 alanylaspartic acid Proteins 0.000 description 1
- 201000009961 allergic asthma Diseases 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 229940125715 antihistaminic agent Drugs 0.000 description 1
- 108010038850 arginyl-isoleucyl-tyrosine Proteins 0.000 description 1
- 210000004507 artificial chromosome Anatomy 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 210000003651 basophil Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000031018 biological processes and functions Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 208000037893 chronic inflammatory disorder Diseases 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000013068 control sample Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 239000003246 corticosteroid Substances 0.000 description 1
- 229960001334 corticosteroids Drugs 0.000 description 1
- 239000007822 coupling agent Substances 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000013480 data collection Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000012470 diluted sample Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 210000000222 eosinocyte Anatomy 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 210000002443 helper t lymphocyte Anatomy 0.000 description 1
- 239000000833 heterodimer Substances 0.000 description 1
- 102000054663 human IL4R Human genes 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 229940088592 immunologic factor Drugs 0.000 description 1
- 239000000367 immunologic factor Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 208000030603 inherited susceptibility to asthma Diseases 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 108040003607 interleukin-13 receptor activity proteins Proteins 0.000 description 1
- 210000002510 keratinocyte Anatomy 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 210000004698 lymphocyte Anatomy 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- NNCAWEWCFVZOGF-UHFFFAOYSA-N mepiquat Chemical compound C[N+]1(C)CCCCC1 NNCAWEWCFVZOGF-UHFFFAOYSA-N 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 229940125645 monoclonal antibody drug Drugs 0.000 description 1
- QJGQUHMNIGDVPM-UHFFFAOYSA-N nitrogen(.) Chemical compound [N] QJGQUHMNIGDVPM-UHFFFAOYSA-N 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 230000009465 prokaryotic expression Effects 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- BHMBVRSPMRCCGG-OUTUXVNYSA-N prostaglandin D2 Chemical compound CCCCC[C@H](O)\C=C\[C@@H]1[C@@H](C\C=C/CCCC(O)=O)[C@@H](O)CC1=O BHMBVRSPMRCCGG-OUTUXVNYSA-N 0.000 description 1
- BHMBVRSPMRCCGG-UHFFFAOYSA-N prostaglandine D2 Natural products CCCCCC(O)C=CC1C(CC=CCCCC(O)=O)C(O)CC1=O BHMBVRSPMRCCGG-UHFFFAOYSA-N 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000010076 replication Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- -1 small molecule compound Chemical class 0.000 description 1
- 150000003384 small molecules Chemical class 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 235000000346 sugar Nutrition 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000004565 tumor cell growth Effects 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 210000000689 upper leg Anatomy 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 230000005924 vaccine-induced immune response Effects 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 210000001631 vena cava inferior Anatomy 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 108010027345 wheylin-1 peptide Proteins 0.000 description 1
Images
Classifications
-
- 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
- C12N15/62—DNA sequences coding for fusion proteins
-
- 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
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/2866—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for cytokines, lymphokines, interferons
-
- 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
- C07K16/24—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against cytokines, lymphokines or interferons
- C07K16/244—Interleukins [IL]
- C07K16/247—IL-4
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
- A61K45/06—Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
- A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
- A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
- A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
- A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/02—Nasal agents, e.g. decongestants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/06—Antiasthmatics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/02—Drugs for skeletal disorders for joint disorders, e.g. arthritis, arthrosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
-
- 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
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
-
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
-
- 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
- C12N5/12—Fused cells, e.g. hybridomas
-
- 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
- A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
- C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
-
- 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
- C07K2317/50—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
- C07K2317/565—Complementarity determining region [CDR]
-
- 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
- C07K2317/567—Framework region [FR]
-
- 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
- C07K2317/00—Immunoglobulins specific features
- C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin
- C07K2317/92—Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
-
- 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
- C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/85—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells
- C12N15/8509—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic
- C12N2015/8518—Vectors or expression systems specially adapted for eukaryotic hosts for animal cells for producing genetically modified animals, e.g. transgenic expressing industrially exogenous proteins, e.g. for pharmaceutical use, human insulin, blood factors, immunoglobulins, pseudoparticles
Abstract
本发明提供一种能够结合白细胞介素4(IL‑4)受体(IL‑4R)的抗体。此外,本发明提供编码所述抗体的核酸序列,包含所述核酸序列的载体以及用所述载体转化或转染的宿主细胞。更进一步地,本发明提供用于生产本发明所述抗体的方法,所述抗体的医学用途以及包含所述抗体的试剂盒。
Description
技术领域
本发明涉及生物制药领域,具体而言,本发明涉及能够结合白细胞介素4(IL-4)受体(IL-4R)的抗体及其用途。
背景技术
白细胞介素-4(IL-4)是主要由活化的T细胞、单核细胞、嗜碱粒细胞、肥大细胞和嗜酸粒细胞产生的细胞因子。IL-4涉及多种生物学过程,已知其生物作用包括刺激活化B细胞和T细胞增殖、CD4+T细胞分化成II型辅助T细胞。研究表明,IL-4在介导过敏性疾病、自身免疫性疾病、感染性疾病、肿瘤等疾病的免疫反应中有多重作用,对肿瘤、自身免疫性疾病和感染性疾病等有治疗作用,并且IL-4对疫苗免疫应答具有调节作用,因此,IL-4一直是人们关注的研究热点之一。
IL-13也是由活化的T细胞分泌产生的一种细胞因子,在不同的细胞类型如单核细胞、B细胞、肥大细胞和角质形成细胞中具有不同功能。IL-13能抑制单核细胞释放炎性细胞因子和化学因子,诱导B细胞增殖和分化,促进IgE合成。IL-13与IL-4在生物功能方面有许多共性,包括抑制单核细胞释放炎症介质,诱导巨噬细胞树突样变,促进单核细胞表面表达CD23及刺激B细胞合成免疫球蛋白。同时IL-13也具有自身的生物学功能特点,主要包括:促进人单核细胞分化及细胞表面抗原发生改变;诱导B细胞增殖,分化,促进B细胞分泌抗体;调节IgE合成,与机体变态反应相关;抑制肿瘤细胞生长;抑制HIV的复制等。
IL-4的生物活性是由特异的细胞表面IL-4受体(IL-4R,在人类中为“hIL-4R”)介导的。人IL-4R是由两条多肽链形成的异二聚体,其中一条α链(hIL-4Rα,UniProtKB:P24394)对IL-4有很高的亲和力。并且研究表明,IL-13的细胞表面受体α链(IL-13Rα链)也和IL-4Rα链共同组成另一种形式的IL-4R复合体。由于在IL-4R复合物中IL-4Rα链对IL-4的结合起主导作用,并且其还涉及其他细胞因子,因此目前人们把IL-4Rα链作为主要对象开展研究,并且针对该靶点的人单克隆抗体已经在临床上证明可以有效缓解和治疗哮喘、湿疹以及特应性皮炎等症状。
已知人白细胞介素-4受体可产生一种可溶形式的蛋白(shIL-4Rα,SEQ IDNO.94),这种可溶形式的蛋白可以抑制IL-4介导的细胞增殖和T细胞介导的IL-5上调。该受体的两种形式与过敏反应相关,其表现为过敏性鼻炎、鼻窦炎、哮喘或湿疹等病症。因此将该蛋白作为靶点的阻断抗体有助于治疗和缓解此类疾病。
哮喘(Asthma)是一种由多种炎症细胞如嗜酸性粒细胞、肥大细胞和淋巴细胞共同参与的慢性气道炎症性疾病,其具体发病机制目前尚不清楚。IL-4等细胞因子在支气管哮喘发生发展中起着十分重要的作用,研制IL-4的特异性抗体是解决哮喘治疗的有效途径之一。抑制IL-4/IL-4Rα可以对哮喘起到有效的免疫调节作用。
过敏性鼻炎(allergic rhinitis,AR)与哮喘的发病机制有许多共同之处,均属于I型变态反应。研究发现,IL-4、IL-17和IgE在过敏性鼻炎的发病机制中扮演了重要的角色。药物治疗是目前AR治疗的重点,其中,鼻用皮质类固醇和抗组胺药物处于核心地位。
特应性皮炎(atopic dermatitis,AD),又称异位性皮炎或遗传过敏性皮炎,是皮肤科常见疾病,多见于儿童和青少年,常与某些遗传过敏性疾病如过敏性鼻炎、哮喘等并发。IL-4和IL-13等细胞因子参与的免疫因素是其主要发病机制之一。
嗜酸粒细胞性食管炎(eosinophilic esophagitis,EoE)是以食管壁全层嗜酸粒细胞(eosinophils,EOS)浸润为特征的慢性免疫性炎性疾病。EoE发病与Th2细胞功能紊乱相关。目前特异性强的方案,如新型生物制剂抗IL-5(如美泊利单抗)成为研究的热点。免疫调节治疗已在动物模型中取得成果,但人类临床试验仍需探索。PGD2抑制剂、抗TNF-α、抗IL-13等药物正在探索中。
目前针对hIL-4R靶点的单克隆抗体药物进入临床试验,如Dupilumab,在治疗特应性皮炎的II期临床中表现出了较好疗效。除Dupilumab以外,亦有公司申请了针对hIL-4R的其他单克隆抗体专利,如US 7,186,809和US 7,638,606。
发明内容
本发明通过抗体筛选和优化获得了针对IL-4R的特异性抗体,所述抗体能够作为IL-4与IL-4R结合的阻断剂,通过与IL-4R结合,可以用于治疗炎症或过敏性疾病等。
优选地,本发明的抗体包含轻链可变区(VL),所述轻链可变区包含选自下述的CDR1、CDR2和CDR3组合:
(1)SEQ ID NO:1所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:5所示的CDR3;
(2)SEQ ID NO:2所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:5所示的CDR3;
(3)SEQ ID NO:2所示的CDR1,SEQ ID NO:4所示的CDR2和SEQ ID NO:5所示的CDR3;
(4)SEQ ID NO:1所示的CDR1,SEQ ID NO.4所示的CDR2和SEQ ID NO:5所示的CDR3;
(5)SEQ ID NO:2所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:6所示的CDR3;
(6)SEQ ID NO:2所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:7所示的CDR3;
(7)SEQ ID NO:2所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:8所示的CDR3;
(8)SEQ ID NO:1所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:6所示的CDR3;
(9)SEQ ID NO:2所示的CDR1,SEQ ID NO:4所示的CDR2和SEQ ID NO:6所示的CDR3;
(10)SEQ ID NO:2所示的CDR1,SEQ ID NO:4所示的CDR2和SEQ ID NO:8所示的CDR3;
(11)SEQ ID NO:1所示的CDR1,SEQ ID NO:4所示的CDR2和SEQ ID NO:8所示的CDR3;和
(12)SEQ ID NO:1所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:8所示的CDR3;
如/或
所述抗体包含重链可变区(VH),所述重链可变区包含选自下述的CDR1、CDR2和CDR3组合:
(1)SEQ ID NO:14所示的CDR1,SEQ ID NO:17所示的CDR2和SEQ ID NO:19所示的CDR3;
(2)SEQ ID NO:14所示的CDR1,SEQ ID NO:18所示的CDR2和SEQ ID NO:19所示的CDR3;
(3)SEQ ID NO:14所示的CDR1,SEQ ID NO:17所示的CDR2和SEQ ID NO:20所示的CDR3;
(4)SEQ ID NO:14所示的CDR1,SEQ ID NO:18所示的CDR2和SEQ ID NO:20所示的CDR3;
(5)SEQ ID NO:15所示的CDR1,SEQ ID NO:17所示的CDR2和SEQ ID NO:19所示的CDR3;
(6)SEQ ID NO:16所示的CDR1,SEQ ID NO:17所示的CDR2和SEQ ID NO:19所示的CDR3;和
(7)SEQ ID NO:14所示的CDR1,SEQ ID NO:18所示的CDR2和SEQ ID NO:19所示的CDR3。
进一步地,所述抗体的轻链可变区包含选自下述的FR1、FR2、FR3和FR4组合:
(1)SEQ ID NO:9所示的FR1,SEQ ID NO:10所示的FR2,SEQ ID NO:12所示的FR3和SEQ ID NO:13所示的FR4;和
(2)SEQ ID NO:9所示的FR1,SEQ ID NO:11所示的FR2,SEQ ID NO:12所示的FR3和SEQ ID NO:13所示的FR4。
进一步地,所述抗体的重链可变区包含选自下述的FR1、FR2、FR3和FR4组合:
(1)SEQ ID NO:21所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(2)SEQ ID NO:22所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(3)SEQ ID NO:23所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(4)SEQ ID NO:24所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(5)SEQ ID NO:24所示的FRl,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(6)SEQ ID NO:25所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(7)SEQ ID NO:26所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(8)SEQ ID NO:27所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(9)SEQ ID NO:29所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(10)SEQ ID NO:30所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(11)SEQ ID NO:24所示的FR1,SEQ ID NO:33所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(12)SEQ ID NO:24所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:36所示的FR3和SEQ ID NO:38所示的FR4;
(13)SEQ ID NO:24所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:37所示的FR3和SEQ ID NO:38所示的FR4;
(14)SEQ ID NO:31所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(15)SEQ ID NO:27所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(16)SEQ ID NO:26所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(17)SEQ ID NO:25所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(18)SEQ ID NO:28所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(19)SEQ ID NO:28所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(20)SEQ ID NO:23所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(21)SEQ ID NO:22所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;和
(22)SEQ ID NO:21所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4。
按照本领域公知的抗体轻链可变区、重链可变区的结构域组成,本发明抗体的轻链可变区或重链可变区以FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4的顺序包含上述结构域组分,或者以(X)n-FR1-(X)n-CDR1-(X)n-FR2-(X)n-CDR2-(X)n-FR3-(X)n-CDR3-(X)n-FR4-(X)n的顺序包含上述结构域组分,其中X为任意氨基酸残基,n为零或大于零的整数。
优选地,本发明提供的抗体包含选自下述序列所示氨基酸序列的轻链可变区:
SEQ ID NO:39、SEQ ID NO:40、SEQ ID NO:41、SEQ ID NO:42、SEQ ID NO:43、SEQID NO:44、SEQ ID NO:45、SEQ ID NO:46、SEQ ID NO:47、SEQ ID NO:48、SEQ ID NO:49、SEQID NO:50、SEQ ID NO:51、SEQ ID NO:52、SEQ ID NO:53、SEQ ID NO:54、SEQ ID NO:55、SEQID NO:56、SEQ ID NO:57和SEQ ID NO:58;
和/或
本发明提供的抗体包含选自下述序列所示氨基酸序列的重链可变区:
SEQ ID NO:59、SEQ ID NO:60、SEQ ID NO:61、SEQ ID NO:62、SEQ ID NO:63、SEQID NO:64、SEQ ID NO:65、SEQ ID NO:66、SEQ ID NO:67、SEQ ID NO:68、SEQ ID NO:69、SEQID NO:70、SEQ ID NO:71、SEQ ID NO:72、SEQ ID NO:73、SEQ ID NO:74、SEQ ID NO:75、SEQID NO:76、SEQ ID NO:77、SEQ ID NO:78、SEQ ID NO:79、SEQ ID NO:80、SEQ ID NO:81、SEQID NO:82、SEQ ID NO:83、SEQ ID NO:84、SEQ ID NO:85、SEQ ID NO:86、SEQ ID NO:87、SEQID NO:88、SEQ ID NO:89、SEQ ID NO:90、SEQ ID NO:91、SEQ ID NO:92和SEQ ID NO:93。
根据本发明的具体实施方式,本发明提供的抗体包含选自下述的轻链可变区和重链可变区组合:
(1)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.62所示的重链可变区:
(2)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.63所示的重链可变区;
(3)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.59所示的重链可变区:
(4)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.60所示的重链可变区;
(5)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.61所示的重链可变区;
(6)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.67所示的重链可变区:
(7)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.65所示的重链可变区;
(8)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.66所示的重链可变区;
(9)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.64所示的重链可变区:
(10)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.68所示的重链可变区;
(11)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.69所示的重链可变区;
(12)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.70所示的重链可变区;
(13)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.71所示的重链可变区;
(14)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.72所示的重链可变区;
(15)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.73所示的重链可变区;
(16)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.89所示的重链可变区;
(17)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.88所示的重链可变区;
(18)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.87所示的重链可变区;
(19)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.86所示的重链可变区;
(20)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.83所示的重链可变区;
(21)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.82所示的重链可变区;
(22)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.81所示的重链可变区;
(23)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.85所示的重链可变区;
(24)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.84所示的重链可变区;
(25)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(26)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.90所示的重链可变区;
(27)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.74所示的重链可变区;
(28)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.75所示的重链可变区;
(29)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.76所示的重链可变区;
(30)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.77所示的重链可变区;
(31)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.78所示的重链可变区;
(32)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.80所示的重链可变区;
(33)SEQ ID NO.39所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(34)SEQ ID NO.40所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(35)SEQ ID NO.41所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(36)SEQ ID NO.42所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(37)SEQ ID NO.43所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(38)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(39)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.62所示的重链可变区;
(40)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.68所示的重链可变区;
(41)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.72所示的重链可变区;
(42)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.82所示的重链可变区;
(43)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.85所示的重链可变区;
(44)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(45)SEQ ID NO.48所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(46)SEQ ID NO.49所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(47)SEQ ID NO.50所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(48)SEQ ID NO.45所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(49)SEQ ID NO.46所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(50)SEQ ID NO.47所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(51)SEQ ID NO.56所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(52)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(53)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.62所示的重链可变区;
(54)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.68所示的重链可变区;
(55)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.72所示的重链可变区;
(56)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.82所示的重链可变区;
(57)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.85所示的重链可变区;
(58)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(59)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(60)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.62所示的重链可变区;
(61)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.68所示的重链可变区;
(62)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.72所示的重链可变区;
(63)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.82所示的重链可变区;
(64)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.85所示的重链可变区;
(65)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(66)SEQ ID NO.53所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(67)SEQ ID NO.51所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(68)SEQ ID NO.52所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(69)SEQ ID NO.52所示的轻链可变区和SEQ ID NO.62所示的重链可变区;和
(70)SEQ ID NO.52所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(71)SEQ ID NO.58所示的轻链可变区和SEQ ID NO.92所示的重链可变区;和
(72)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.93所示的重链可变区。
本发明提供的抗体能够结合IL-4R,作为IL-4R的拮抗剂发挥作用。优选地,所述抗体能够结合IL-4Rα,优选结合哺乳动物类IL-4Rα,更优选结合人IL-4Rα,甚至更优选结合人可溶性IL-4Rα。
可以通过Biacore或ELISA方法测定本发明提供的抗体与IL-4Rα的结合亲和力。测得所述抗体能够以小于100nM、小于10nM、小于1nM、小于0.5nM和甚至小于0.1nM亲和力结合IL-4Rα。
在相同条件下,本发明提供的抗体与参照抗体的表达量比为0.1-3∶1,优选0.3-3∶1,更优选0.4-3∶1,更优选0.5-3∶1,更优选0.6-3∶1,更优选0.7-3∶1,更优选1-3∶1。
就抗体类别而言,本发明提供的抗体可以是单克隆抗体、完全或部分人源化抗体或嵌合抗体;
或者优选地,所述抗体为免疫球蛋白,优选为IgA、IgD、IgE、IgG或IgM,更优选为IgG1、IgG2、IgG3或IgG4亚型,更优选为IgG2或IgG4亚型。
另一方面,本发明提供一种融合蛋白或偶联物,所述融合蛋白或偶联物包含本发明所述的抗体。该融合蛋白或偶联物还包含通过化学或物理方法结合于本发明所述抗体的细胞表面受体、活性蛋白及多肽、小分子化合物如氨基酸和糖类、小分子聚合物或对本发明所述抗体进行化学修饰的其它部分等。
又一方面,本发明提供一种核酸序列,所述核酸序列能够编码本发明所提供抗体的重链可变区和/或轻链可变区;
优选地,所述核酸序列能够编码本发明提供抗体的重链和/或轻链。
再一方面,本发明还提供一种包含本发明提供的核酸序列的载体。所述载体可以为真核表达载体、原核表达载体、人工染色体及噬菌体载体等。
上述载体或核酸序列可以用于转化或转染宿主细胞,用于保存或抗体表达等目的。因此,本发明还提供一种采用所述核酸序列或载体转化或转染的宿主细胞。宿主细胞可以是任何原核或真核细胞,例如细菌或昆虫、真菌、植物或动物细胞。
本发明提供的抗体可以利用本领域已知的任何方法获得。例如,可以先由本发明提供的核酸序列获得所述抗体的重链可变区和/或轻链可变区,或者获得所述抗体的重链和/或轻链,然后与所述抗体的任选其他结构域组装成抗体;或者,在允许本发明提供的宿主细胞表达所述抗体的重链可变区和/或轻链可变区或者所述抗体的重链和/或轻链以组装成所述抗体的情况下,培养所述宿主细胞。
任选地,所述方法还包括回收产生的抗体的步骤。
本发明提供的抗体、融合蛋白或偶联物、核酸序列、载体、宿主细胞或者通过上述方法生产的抗体可以被包含在药物组合物中,更特别地被包含在药物制剂中,从而根据实际需要用于各种目的。因此,在又一方面,本发明还提供一种药物组合物,所述药物组合物包含本发明所述的抗体、融合蛋白或偶联物、核酸序列、载体、宿主细胞和/或通过上述方法生产的抗体。
可选地,所述药物组合物可以是药物制剂。所述药物制剂例如为注射剂剂型。
取决于特定剂型,所述药物组合物或药物制剂还包含药学上可接受的载体或赋形剂。
在药物组合物或药物制剂中,还可以包含至少一种选自下述的药物:平喘药如沙丁胺醇等,抗组胺药如氯雷他定等,免疫抑制剂如他克莫司、吡美莫司等,M受体阻断剂如异丙托溴铵等,白三烯受体阻断药如孟鲁司特等,磷酸二酯酶抑制剂如茶碱等,非甾体抗炎药如5-氨基水杨酸等,激素类如倍氯米松,布地奈德等,即本发明提供的抗体、融合蛋白或偶联物、核酸序列、载体、宿主细胞或者通过上述方法生产的抗体可以根据需要与其他药物联合使用。
再一方面,本发明还提供所述抗体、融合蛋白或偶联物、核酸序列、载体、宿主细胞和/或通过所述方法生产的抗体在制备用于预防、治疗或改善炎症或过敏性疾病的药物中的用途;
优选地,所述炎症或过敏性疾病包括自身免疫学疾病,例如过敏性皮炎、哮喘、嗜酸性粒细胞食管炎、湿疹、过敏性鼻炎、鼻息肉、类风湿性关节炎等。
还一方面,本发明提供一种预防、治疗或改善受试者的炎症或过敏性疾病的方法,所述方法包括给有此需要的受试者施用本发明提供的抗体、融合蛋白或偶联物、核酸序列、载体、宿主细胞和/或通过所述方法生产的抗体。
优选地,所述受试者为哺乳类动物;更优选地,所述受试者为人。
优选地,所述炎症或过敏性疾病包括自身免疫学疾病,例如过敏性皮炎、哮喘、嗜酸性粒细胞食管炎、湿疹、过敏性鼻炎、鼻息肉、类风湿性关节炎等。
在预防、治疗或改善炎症或过敏性疾病时,还可以联合应用其他药物,例如所述方法还包括给受试者施用至少一种选自下述的药物:平喘药如沙丁胺醇等,抗组胺药如氯雷他定等,免疫抑制剂如他克莫司、吡美莫司等,M受体阻断剂如异丙托溴铵等,白三烯受体阻断药如孟鲁司特等,磷酸二酯酶抑制剂如茶碱等,非甾体抗炎药如5-氨基水杨酸等,激素类如倍氯米松、布地奈德等。
优选地,所述药物与本发明提供的抗体、融合蛋白或偶联物、核酸序列、载体、宿主细胞和/或通过所述方法生产的抗体同时或顺序施用。
再一方面,本发明还提供一种试剂盒,所述试剂盒包括本发明提供的抗体、融合蛋白或偶联物、核酸序列、载体、宿主细胞和/或通过所述方法生产的抗体。
附图说明
结合本发明非限制性实施例和附图可以更好地理解本发明,在附图中:
图1示出了本发明的抗体在小鼠体内的药代动力学曲线;
图2示出了本发明的抗体在食蟹猴体内的药代动力学曲线;
图3A-3C通过FACS示出了本发明的抗体与表达IL-4Rα的TF-1细胞特异性结合,其中3A示出没有加入本发明抗体L1012H1031时的荧光信号,3B示出加入本发明抗体L1012H1031后的荧光信号,3C示出3A-3B的信号叠加比较。
图4A-4D通过FACS示出了本发明的抗体与表达IL-4Rα的TF-1细胞的特异性结合被体系中存在的sIL-4Rα所阻断,其中4A示出没有加入本发明抗体L1012H1031时的荧光信号,4B示出加入本发明抗体L1012H1031后的荧光信号,4C示出加入本发明抗体L1012H1031及sIL-4Rα时的荧光信号,4D示出4A-4C的信号叠加比较。
具体实施方式
以下参照具体的实施例来说明本发明。本领域技术人员能够理解,这些实施例仅用于说明本发明,其不以任何方式限制本发明的范围。
下述实施例中的实验方法如无特殊说明,均为常规方法;下述实施例中所用的试验材料,如无特殊说明,均为自常规生化试剂商店购买得到的。
在下述实施例中,示例性地提供并验证了表1中所示抗体的作用。
表1本发明提供的示例性抗体
实施例1:本发明抗体的制备与非还原SDS-PAGE凝胶电泳法测定表达量
将抗体轻链可变区的编码序列利用EcoRI和BsiWI酶切位点插入到pFUSE2ss-CLIg-hK载体(Invivogen,货号:pfuse2ss-hclk)构成轻链表达载体;重链可变区的编码序列利用EcoRI和NheI酶切位点插入到pFUSEss-CHIg-hG2载体(Invivogen,货号:pfusess-hchg2)或pFUSEss-CHIg-hG4载体(Invivogen,货号:pfusess-hchg4)构成重链表达载体。
Expi293细胞的培养和转染按照Invitrogen公司Expi293TM Expression SystemKit手册进行(货号:A14635)。转染时细胞密度调整为2×106个/ml,每毫升细胞加入上述0.6μg轻链表达载体和0.4μg重链表达载体,四天后收集培养上清。
细胞培养上清按照《分子克隆实验指南》第三版附录8所述方法进行非还原SDS-PAGE凝胶电泳。
使用东方君意凝胶扫描成像系统拍照并使用Gel-PRO ANALYZER软件进行凝胶定量,以测定抗体瞬转的表达量。结果以相对于对照抗体1(根据专利US 7,186,809构建,其轻链可变区为US 7,186,809的SEQ ID No.10,重链可变区为US 7,186,809的SEQ ID No.12,下同)的表达量表示(对照抗体2根据专利US 7,638,606构建,其轻链可变区为US 7,638,606的SEQ ID No.6,重链可变区为US 7,638,606的SEQ ID No.42,下同),见下表2a-2c。
表2a本发明抗体的瞬转表达量(表达量较对照抗体1显著提高的抗体):
表2b本发明抗体的瞬转表达量(表达量较对照抗体1略有降低的抗体):
表2c本发明抗体的瞬转表达量(表达量较对照抗体1有大幅降低的抗体):
实施例2:检测本发明的抗体抑制hIL-4或hIL-13对TF-1细胞的增殖作用
1.试剂的配制
hIL-4(Invivogen,货号:rhil-4)溶液:用100μl含0.1%BSA(Beyotime,货号:ST023)的PBS溶液溶解hIL-4,得到100μg/ml的浓度,将溶解后的hIL-4按照5μl每管的体积分装至1.5ml(Nunc)离心管内,并存放于-20℃冰箱内。
WST-1(Beyotime,货号:C0036)溶液:把5ml电子耦合剂(C0036-2)加入到WST-1粉末(C0036-1)中,完全溶解即成WST-1溶液,按照每管620μl体积分装至1.5ml离心管内,并存放于-20℃冰箱内。
2.TF-1细胞培养
取出液氮中冻存的TF-1(ATCC:CRL-2003TM)细胞,在37℃水浴中震荡,使其迅速溶解。将溶解后的细胞悬液移至15ml离心管内,加1640培养基至10ml,800rpm离心5min,吸去上清,保留细胞沉淀,重复洗涤一次,加入10ml含10%FBS和2ng/ml GM-CSF(SinoBiological,货号:10015-H01H)的1640培养基,调整细胞密度为1×105~1×106个/ml,移至T75细胞培养瓶(Nunc)中,置于37℃,5%CO2培养箱(Thermo)中静置培养。每隔2-3天,取细胞悬液,800rpm离心5min,用10ml培养基重悬细胞,计数细胞1×106个细胞移入新的T75细胞瓶中,同时补加培养基至10ml,连续传代2-3次,至细胞状态良好(细胞透亮,单一悬浮的略微不规则形态细胞)后可进行增殖实验。
3.抗体的制备和纯化
a)本发明抗体细胞培养上清样品:
将带有不同组抗体基因的质粒转染Expi293细胞,转染后4天取细胞培养上清200μl,800rpm离心5min,将上清经0.22μm孔径滤膜过滤后,用于增殖阻断实验。
b)纯化的本发明抗体样品:取表达本发明抗体的细胞培养上清,过0.22μm滤膜,经GE MabSelect Sure(Cat Number:11003494)Protein A亲和层析柱纯化,纯化系统为GEAKTA purifier 10,纯化后收集的抗体使用Amicon超滤浓缩管(Cat Number:UFC903096)浓缩并定量。检测时,用PBS稀释抗体至0至1μg/ml用于增殖阻断实验。
4.抑制增殖实验
取T75细胞瓶中生长状态良好细胞,移入15ml离心管内,800rpm离心5min,弃上清,取细胞沉淀。用10ml PBS重悬细胞,800rpm离心5min,弃上清,取细胞沉淀。用10ml含10%FBS的1640培养基(不含GM-CSF)重悬细胞,800rpm离心5min,弃上清,取细胞沉淀。用5ml含10%FBS的1640培养基(不含GM-CSF)重悬细胞,细胞计数后,补加培养基,调整细胞密度至5×105个/ml。将细胞悬液按每孔80μl体积加入96孔板内(预留外圈防挥发孔不加细胞)。将准备好的不同浓度的纯化抗体10μl或细胞培养上清10μl加入96孔的细胞内(3个重复孔)。然后用含10%FBS的1640培养基稀释hIL-4至50ng/ml。将50ng/ml的hIL-4按照10μl每孔的体积分别加入96孔板上对应的细胞孔内,使得最终的细胞密度为4×105个/ml,hIL-4浓度为5ng/ml,最终96孔板每孔体积为100μl,设置不添加hIL-4,不添加抗体,仅加入相同数量细胞与同体积培养液的阴性对照组(3个重复孔),另设置不添加抗体,补加相同体积培养液,细胞中仅加入相同浓度hIL-4的阳性对照组(3个重复孔)。在96孔细胞板外圈每孔加入200μl PBS防止内圈液体挥发。将96孔细胞板置于37℃,5%CO2培养箱中静置培养。
使用500ng/ml的hIL-13(加入细胞后的终浓度为50ng/ml),按照相同过程重复上述实验。
5.数据统计
96孔细胞板在5%CO2培养箱中静置培养72h后,在每孔细胞内加入10μl WST-1溶液。将96孔细胞板继续置于37℃,5%CO2培养箱中静置培养。24h后,将96孔板置于flexstation 3(Molecular Devices)中,读取OD450-OD650的值。
对于本发明抗体培养上清,测得的OD450-OD650数值(OD值)与阳性对照组和阴性对照组的数值进行计算,抑制百分率=(转染后细胞上清OD值-阳性对照组OD值)/(阴性对照组OD值-阳性对照组OD值)×100%。本发明的抗体对hIL-4或hIL-13的TF-1细胞增殖作用的阻断作用结果见下表3a-3b。
对于经过纯化的不同浓度的本发明抗体,测得的OD450-OD650数据输入prism5软件中,将阴性对照组数值设为最低值,阳性对照组数值设为最高值,抗体浓度取对数值,由prism5软件拟合抗体浓度对数值与OD450-OD650的曲线,计算得到的IC50见下表4。
表3a本发明的抗体抑制TF-1细胞增殖活性的筛选结果(抑制率较对照抗体1增加的抗体)
表3b本发明的抗体抑制TF-1细胞增殖活性的筛选结果(抑制率较对照抗体1等同或降低的抗体)
表4抗体600-900ml瞬转纯化产物活性数据
实施例3:ELISA检测本发明的抗体与sIL-4Rα的结合能力
1.试剂的配制
sIL-4Rα(PEPRO TECH,货号:200-04R)溶液:取一支sIL-4Rα,加入1ml ddH2O,上下颠倒混匀,即为100μg/ml溶液,然后分装后于-20℃冰箱中保存。
待测样品:取瞬染后表达本发明抗体的Expi293细胞培养物上清液(培养基为Expi293 Expression Medium,Invitrogen,货号:A1435102;在8%CO2培养箱中,连续100rpm悬浮培养4天)10μl和2μl,分别加至990μl和998μl的PBS溶液中,配制成为1∶100和1∶500稀释的待测抗体样品。
对照样品:正常细胞培养上清(Expi293细胞不经过转染,培养基为Expi293Expression Medium(Invitrogen,货号:A1435102;在8%CO2培养箱中,连续100rpm悬浮培养4天)同样进行1∶100和1∶500稀释,作为阴性对照样品。
2.ELISA检测
取100μl 100μg/ml sIL-4Rα溶液,加入到9.90ml PBS包被液中,上下颠倒混匀,即为1.0μg/ml抗原包被液。将配好的抗原包被液加入到96孔酶标板(corning)中,每孔100μl。将96孔酶标板置于4℃冰箱中孵育过夜。第二天,弃去其中溶液,向96孔酶标板中逐排加入含2%BSA的PBS溶液,每孔300μl。4℃冰箱中孵育2h。弃去2%BSA,使用PBST洗涤3次。将稀释好的待测抗体依次加入到相应的孔中,同时加入正常细胞培养上清作为阴性对照样品,每个样品做三个复孔,每孔100μl。将酶标板用保鲜膜包裹(或加盖)后,置于10℃恒温培养箱中孵育1h。将96孔酶标板取出,弃去其中溶液,PBST洗涤3次后向96孔酶标板中逐排加入TMB溶液(Solarbio,Cat Number:PR1200),每孔100μl。室温放置5分钟后,立即向96孔酶标板中加入2M H2SO4溶液终止反应。将96孔酶标板置于flexstation 3(Molecular Devices)中,读取OD450的值,数据收集计算分析。结果以相对于对照抗体1的亲和力示出,见下表5a-5c。
表5a本发明的抗体对sIL-4Rα的亲和力(亲和力显著大于对照抗体1的抗体)
抗体编号 | OD450/OD450<sub>对照抗体1</sub> |
对照抗体1 | 1 |
L1021H1000 | 2.42 |
L1020H1000 | 2.27 |
L1019H1000 | 1.79 |
L1001H1000 | 1.56 |
L1012H1000 | 1.22 |
L1000H1031 | 1.14 |
L1020H1031 | 1.12 |
L1000H1014 | 1.06 |
L1020H1029 | 1.01 |
表5b本发明的抗体对sIL-4Rα的亲和力(亲和力较对照抗体1等同或略有降低的抗体)
抗体编号 | OD450/OD450<sub>对照抗体1</sub> |
L1010H1000 | 1 |
L1021H1029 | 1 |
L1011H1000 | 0.9 |
L1008H1000 | 0.9 |
L1021H1031 | 0.9 |
L1024H1031 | 0.9 |
L1007H1000 | 0.8 |
L1020H1016 | 0.8 |
L1000H1029 | 0.8 |
L1000H1001 | 0.7 |
表5c本发明的抗体对sIL-4Rα的亲和力(亲和力较对照抗体1显著降低的抗体)
实施例4:本发明的抗体在小鼠体内的药代动力学
为了进一步筛选抗体,在小鼠体内进行了一系列的药代动力学实验。
选择6-8周龄SPF级Balb/c小鼠,以5mg(本发明的抗体或对照抗体2)/kg(小鼠体重)的量进行抗体皮下注射。于药前(0h),药后2、8、24、48、72、120、168、216、264、336h的时间点采集血样。取样为动物用异氟烷吸入麻醉后,眼眶静脉丛取血,每只取血量约0.1mL;药后336h动物首先用异氟烷吸入麻醉,下腔静脉取血安乐死。
血样不抗凝,取血后2h之内室温1500g离心10min以分离血清。收集的上清液立即转移至新的标记好的离心管,血清保存于-70℃以下暂存。
使用ELISA方法测定小鼠体内的抗体浓度:
1.试剂的配制
sIL-4Rα(PEPRO TECH,货号:200-04R)溶液:取一支sIL-4Rα,加入1ml ddH2O,上下颠倒混匀,即为100μg/ml溶液,然后分装后于-20℃冰箱中保存。
待测样品:取不同时间段采集的血清各1μl,加入至999μl含1%BSA的PBS溶液中,即成1∶1000稀释的待测血清样品。
标准品:将待检测抗体用含1%BSA和0.1%正常动物血清(Beyotime,货号:ST023)的PBS溶液稀释成0.1μg/ml。分别取0.1μg/ml的待检测抗体800、600、400、200、100、50、10、0μl,分别加入含1%BSA和0.1%正常动物血清的PBS溶液200、400、600、800、900、950、990、1000μl,配成最终浓度分别为80、60、40、20、10、5、1、0ng/ml的本发明抗体标准品。
2.ELISA检测
取250μl 100μg/ml sIL-4Rα溶液,加入到9.75ml PBS包被液中,上下颠倒混匀,即为2.5μg/ml抗原包被液。将配好的抗原包被液加入入96孔酶标板(corning)中,每孔100μl。将96孔酶标板用保鲜膜包裹(或加盖)后,4℃冰箱中孵育过夜。第二天,将96孔酶标板取出,弃去其中溶液,加入含2%BSA的PBS溶液,每孔300μl。将酶标板用保鲜膜包裹(或加盖)后,于4℃冰箱中孵育2h。将96孔酶标板取出,弃去其中溶液,PBST重复洗涤3次。将稀释好的标准抗体及待测血清样品依次加入到相应的孔中,每个样品做三个复孔,每孔100μl。将酶标板用保鲜膜包裹(或加盖)后,室温孵育1h。弃去其中溶液,PBST洗涤3次。向96孔酶标板中逐排加入TMB溶液(Solarbio,Cat Number:PR1200),每孔100μl。室温放置5分钟,立即向96孔酶标板中加入2M H2SO4溶液终止反应。将96孔酶标板置于flexstation 3(MolecularDevices)中,读取OD450的值,收集数据使用Winnonlin软件计算结果。药代动力学结果见图1与下表6。
实施例5:本发明的抗体在食蟹猴体内的药代动力学
为了进一步筛选抗体,在食蟹猴体内进行了一系列的药代动力学实验。
选择3-5岁食蟹猴,体重2-5Kg,以5mg(本发明的抗体或对照抗体2)/kg(食蟹猴体重)的量进行抗体皮下注射。用一次性无菌注射器准确抽取要施用的抗体或对照抗体2,于动物大腿内侧皮下进行多点注射,每点注射容量不超过2ml。于药前(0h),药后0.5、2、4、8、24、48、72、120、168、240、336h、432h、504h、600h、672h的时间点从动物后肢皮下静脉采集全血,每只取血量1.5ml。
血样不抗凝,取血后2h之内室温1500g离心10min以分离血清。收集的上清液立即转移至新的标记好的离心管,血清保存于-70℃以下暂存。采用实施例4所述方法测定食蟹猴体内的抗体浓度。药代动力学结果见图2与下表7。
实施例6:FACS检测本发明的抗体与TF-1细胞的结合
1.细胞培养
在液氮中取出冻存的TF-1(ATCC:CRL-2003TM)细胞,在37℃水浴中不停轻柔震荡,使其迅速溶解。加溶解后的细胞悬液移至15ml离心管内,加1640培养基(Hyclone,货号:SH30809.01B)至10ml,800rpm离心5min,吸去上清,保留细胞沉淀,重复洗涤一次,加入含10%FBS(Hyclone,货号:SV30184.02)、2ng/ml GM-CSF(Sino Biological,货号:10015-H01H)的1640培养基,调整细胞密度为1×105-1×106个/ml,移至T75细胞培养瓶(Nunc)中,置于37℃,5%CO2培养箱(Thermo)中静置培养。每隔2-3天,取细胞悬液,800rpm离心5min,用10ml培养基重悬细胞,计数细胞1×106个细胞移入新的T75细胞瓶中,同时补加培养基至10ml,连续传代2-3次,至细胞状态良好(细胞透亮,单一悬浮的略微不规则形态细胞)后可进行实验。
2.细胞处理
取TF-1细胞,通过显微镜计数,将细胞分为3组,分别置于3个1.5ml离心管内,每组细胞个数为1x106个。800rpm 5min离心细胞,用冷的含1%BSA的PBS 1ml重悬细胞,重复洗涤一次。800rpm 5min离心细胞,每个离心管内加入冷的含1%BSA的PBS 45μl重悬细胞后,其中第一个离心管内加入本发明L1012H1031抗体5μl(500μg/ml),第二个离心管内加入PBS5μl作为阴性对照。冰上静置45min后,800rpm 5min离心细胞,用冷的含1%BSA的PBS 1ml重悬细胞,重复洗涤一次。800rpm 5min离心细胞,每个离心管内加入冷的含1%BSA的PBS 499μl重悬细胞后,加入1μl FITC标记山羊抗人IgG(H+L)(Beyotime,货号:A0556)。冰上静置45min后,800rpm 5min离心细胞,用冷的含1%BSA的PBS 1ml重悬细胞,重复洗涤一次,细胞待测。
3.FACS上机检测
按正确流程打开仪器和操作系统,设置正确的参数后,将细胞加入检测管内,检测FL1通道的荧光信号,使用FlowJo 7.6软件进行分析,结果见图3A-3C,显示本发明的抗体能够与表达IL-4Rα的TF-1细胞特异性结合。
实施例7:FACS检测可溶性hIL-4Rα(sIL-4Rα)对抗体与TF-1细胞结合的阻断作用
1.试剂的配制
按照实施例1中所述进行hIL-4(Invivogen,货号:rhil-4)溶液的配制。
2.细胞培养
按照实施例6中所述进行TF-1(ATCC:CRL-2003TM)的细胞培养。
3.sIL-4Rα与本发明的抗体的混合
取sIL-4Rα10μl(100μg/ml)与本发明的L1012H1031抗体5μl(500μg/ml)以2∶1的摩尔比在1.5ml离心管内混合均匀。同时取PBS 10μl与本发明L1012H1031抗体5μl混合作为阳性对照,取PBS 15μl作为阴性对照。将离心管放37℃培养箱内静置1h。
4.细胞处理
取TF-1细胞,通过显微镜计数,将细胞分为4组,分别置于3个1.5ml离心管内,每组细胞个数为1×106个。800rpm 5min离心细胞,用冷的含1%BSA的PBS 1ml重悬细胞,重复洗涤一次。800rpm 5min离心细胞,每个离心管内加入冷的含1%BSA的PBS 35μl重悬细胞后,加入步骤3中不同抗体混合物与对照15μl。冰上静置45min后,800rpm 5min离心细胞,用冷的含1%BSA的PBS 1ml重悬细胞,重复洗涤一次。800rpm 5min离心细胞,每个离心管内加入冷的含1%BSA的PBS 499μl重悬细胞后,加入1μl FITC标记山羊抗人IgG(H+L)(Beyotime,货号:A0556)。冰上静置45min后,800rpm 5min离心细胞,用冷的含1%BSA的PBS 1ml重悬细胞,重复洗涤一次,细胞待测。
5.FACS上机检测
按正确流程打开仪器和操作系统,设置正确的参数后,将细胞加入检测管内,检测FL1通道的荧光信号,使用FlowJo 7.6软件进行分析,结果见图4A-4D,显示sIL-4Rα能够特异性地有效阻断本发明抗体与TF-1细胞的特异性结合。
实施例8:Bicore抗体亲和力测定
首先将抗人Fc(AHC)抗体(GE Healthcare)偶联到CM5芯片上,然后用AHC捕获本发明抗体,再将不同浓度的人sIL-4Rα流过捕获了本发明抗体的芯片表面,其中本发明抗体稀释到2μg/ml,抗原sIL-4Rα稀释至0.39、0.78、1.56、3.13、6.25、12.5、25.0、50.0和100.0nM。然后在Biacore T200 control software软件中建立实验方法,之后运行实验程序检测。结果见下表8。
表8抗体亲和力检测结果
序列表
<110> 苏州康乃德生物医药有限公司
<120> 用于结合白细胞介素4受体的抗体
<130> LC16110057-D
<160> 94
<170> PatentIn version 3.3
<210> 1
<211> 12
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LCDR1-1
<400> 1
Arg Ala Ser Gln Ser Val Ser Asn Ser Tyr Leu Ala
1 5 10
<210> 2
<211> 12
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LCDR1-2
<400> 2
Arg Ala Ser Gln Ser Val Ser Ser Ser Tyr Leu Ala
1 5 10
<210> 3
<211> 7
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LCDR2-1
<400> 3
Gly Ala Ser Ser Arg Ala Thr
1 5
<210> 4
<211> 7
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LCDR2-2
<400> 4
Gly Ala Ser Ser Arg Ala Pro
1 5
<210> 5
<211> 10
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LCDR3-1
<400> 5
Gln Gln Tyr Gly Ser Ser Pro Pro Trp Thr
1 5 10
<210> 6
<211> 10
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LCDR3-2
<400> 6
Gln Gln Tyr Asp His Ser Pro Pro Trp Thr
1 5 10
<210> 7
<211> 10
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LCDR3-3
<400> 7
Gln Gln Tyr Gly Ser Ser Ala Gly Trp Thr
1 5 10
<210> 8
<211> 10
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LCDR3-4
<400> 8
Gln Gln Tyr Asp His Ser Ala Gly Trp Thr
1 5 10
<210> 9
<211> 23
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LFR1-1
<400> 9
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys
20
<210> 10
<211> 15
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LFR2-1
<400> 10
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Phe
1 5 10 15
<210> 11
<211> 15
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LFR2-2
<400> 11
Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu Ile Tyr
1 5 10 15
<210> 12
<211> 32
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LFR3-1
<400> 12
Gly Ile Pro Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr
1 5 10 15
Leu Thr Ile Ser Arg Leu Glu Pro Glu Asp Phe Ala Val Tyr Tyr Cys
20 25 30
<210> 13
<211> 10
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> LFR4-1
<400> 13
Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
1 5 10
<210> 14
<211> 5
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HCDR1-1
<400> 14
Arg Asn Ala Met Phe
1 5
<210> 15
<211> 5
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HCDR1-2
<400> 15
Arg Asn Ala Met His
1 5
<210> 16
<211> 5
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HCDR1-3
<400> 16
Asp Tyr Ala Met Phe
1 5
<210> 17
<211> 17
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HCDR2-1
<400> 17
Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys Gly
1 5 10 15
Arg
<210> 18
<211> 17
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HCDR2-2
<400> 18
Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys Gly
1 5 10 15
Arg
<210> 19
<211> 7
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HCDR3-1
<400> 19
Gly Arg Tyr Tyr Phe Asp Tyr
1 5
<210> 20
<211> 7
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HCDR3-2
<400> 20
Gly Arg Tyr Tyr Phe Pro Trp
1 5
<210> 21
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-1
<400> 21
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser
20 25 30
<210> 22
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-2
<400> 22
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser
20 25 30
<210> 23
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-3
<400> 23
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
20 25 30
<210> 24
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-4
<400> 24
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser
20 25 30
<210> 25
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-5
<400> 25
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser
20 25 30
<210> 26
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-6
<400> 26
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
20 25 30
<210> 27
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-7
<400> 27
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
20 25 30
<210> 28
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-8
<400> 28
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser
20 25 30
<210> 29
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-9
<400> 29
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser
20 25 30
<210> 30
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-10
<400> 30
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Thr Cys Ala Gly Ser Gly Phe Thr Phe Ser
20 25 30
<210> 31
<211> 30
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR1-11
<400> 31
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Asp
20 25 30
<210> 32
<211> 14
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR2-1
<400> 32
Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val Ser
1 5 10
<210> 33
<211> 14
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR2-2
<400> 33
Trp Val Arg Gln Ala Pro Gly Glu Gly Leu Glu Trp Val Ser
1 5 10
<210> 34
<211> 31
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR3-1
<400> 34
Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met
1 5 10 15
Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 35
<211> 31
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR3-2
<400> 35
Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met
1 5 10 15
Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 36
<211> 31
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR3-3
<400> 36
Phe Thr Ile Ser Arg Asp Glu Ala Lys Asn Ser Leu Tyr Leu Gln Met
1 5 10 15
Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 37
<211> 31
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR3-4
<400> 37
Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu Gln Met
1 5 10 15
Asn Ser Leu Arg Ala Gly Asp Met Ala Val Tyr Tyr Cys Ala Arg
20 25 30
<210> 38
<211> 11
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> HFR4-1
<400> 38
Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser
1 5 10
<210> 39
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1007)
<400> 39
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 40
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1008)
<400> 40
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 41
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1009)
<400> 41
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 42
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1010)
<400> 42
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 43
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1011)
<400> 43
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 44
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1012)
<400> 44
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 45
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1016)
<400> 45
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 46
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1017)
<400> 46
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 47
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1018)
<400> 47
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 48
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1013)
<400> 48
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 49
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1014)
<400> 49
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 50
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1015)
<400> 50
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 51
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1023)
<400> 51
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 52
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1024)
<400> 52
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 53
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1022)
<400> 53
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 54
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1021)
<400> 54
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 55
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1020)
<400> 55
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 56
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1019)
<400> 56
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 57
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1000)
<400> 57
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Ser Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Phe Gly Ala Ser Ser Arg Ala Thr Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Gly Ser Ser Pro
85 90 95
Pro Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 58
<211> 109
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VL(L1001)
<400> 58
Glu Ile Val Leu Thr Gln Ser Pro Gly Thr Leu Ser Leu Ser Pro Gly
1 5 10 15
Glu Arg Ala Thr Leu Ser Cys Arg Ala Ser Gln Ser Val Ser Asn Ser
20 25 30
Tyr Leu Ala Trp Tyr Gln Gln Lys Pro Gly Gln Ala Pro Arg Leu Leu
35 40 45
Ile Tyr Gly Ala Ser Ser Arg Ala Pro Gly Ile Pro Asp Arg Phe Ser
50 55 60
Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Arg Leu Glu
65 70 75 80
Pro Glu Asp Phe Ala Val Tyr Tyr Cys Gln Gln Tyr Asp His Ser Ala
85 90 95
Gly Trp Thr Phe Gly Gln Gly Thr Lys Val Glu Ile Lys
100 105
<210> 59
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1009)
<400> 59
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 60
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1010)
<400> 60
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 61
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1011)
<400> 61
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 62
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1007)
<400> 62
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 63
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1008)
<400> 63
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 64
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1015)
<400> 64
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 65
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1013)
<400> 65
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 66
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1014)
<400> 66
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 67
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1012)
<400> 67
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 68
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1016)
<400> 68
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 69
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1017)
<400> 69
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 70
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1018)
<400> 70
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 71
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1019)
<400> 71
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 72
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1020)
<400> 72
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 73
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1021)
<400> 73
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 74
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1033)
<400> 74
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Arg
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 75
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1034)
<400> 75
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Thr Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 76
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1035)
<400> 76
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met His Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 77
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1036)
<400> 77
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Glu Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 78
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1037)
<400> 78
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Glu Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 79
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1039)
<400> 79
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Gly Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 80
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1038)
<400> 80
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Asp Asp Tyr
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 81
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1028)
<400> 81
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 82
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1027)
<400> 82
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 83
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1026)
<400> 83
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 84
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1030)
<400> 84
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 85
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1029)
<400> 85
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 86
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1025)
<400> 86
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 87
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1024)
<400> 87
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 88
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1023)
<400> 88
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 89
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1022)
<400> 89
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 90
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1032)
<400> 90
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 91
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1031)
<400> 91
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 92
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1000)
<400> 92
Glu Val Gln Leu Val Gln Ser Gly Gly Gly Leu Val His Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Gly Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Asn Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Met Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 93
<211> 115
<212> PRT
<213> 人工序列(Artificial sequence)
<220>
<223> VH(H1001)
<400> 93
Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly
1 5 10 15
Ser Leu Arg Leu Ser Cys Ala Ala Ser Gly Phe Thr Phe Ser Arg Asn
20 25 30
Ala Met Phe Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val
35 40 45
Ser Gly Ile Gly Thr Gly Gly Ala Thr Ser Tyr Ala Asp Ser Val Lys
50 55 60
Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Ser Leu Tyr Leu
65 70 75 80
Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys Ala
85 90 95
Arg Gly Arg Tyr Tyr Phe Pro Trp Trp Gly Gln Gly Thr Leu Val Thr
100 105 110
Val Ser Ser
115
<210> 94
<211> 209
<212> PRT
<213> 人(human)
<220>
<223> sIL4Rα
<400> 94
Gly Asn Met Lys Val Leu Gln Glu Pro Thr Cys Val Ser Asp Tyr Met
1 5 10 15
Ser Ile Ser Thr Cys Glu Trp Lys Met Asn Gly Pro Thr Asn Cys Ser
20 25 30
Thr Glu Leu Arg Leu Leu Tyr Gln Leu Val Phe Leu Leu Ser Glu Ala
35 40 45
His Thr Cys Ile Pro Glu Asn Asn Gly Gly Ala Gly Cys Val Cys His
50 55 60
Leu Leu Met Asp Asp Val Val Ser Ala Asp Asn Tyr Thr Leu Asp Leu
65 70 75 80
Trp Ala Gly Gln Gln Leu Leu Trp Lys Gly Ser Phe Lys Pro Ser Glu
85 90 95
His Val Lys Pro Arg Ala Pro Gly Asn Leu Thr Val His Thr Asn Val
100 105 110
Ser Asp Thr Leu Leu Leu Thr Trp Ser Asn Pro Tyr Pro Pro Asp Asn
115 120 125
Tyr Leu Tyr Asn His Leu Thr Tyr Ala Val Asn Ile Trp Ser Glu Asn
130 135 140
Asp Pro Ala Asp Phe Arg Ile Tyr Asn Val Thr Tyr Leu Glu Pro Ser
145 150 155 160
Leu Arg Ile Ala Ala Ser Thr Leu Lys Ser Gly Ile Ser Tyr Arg Ala
165 170 175
Arg Val Arg Ala Trp Ala Gln Cys Tyr Asn Thr Thr Trp Ser Glu Trp
180 185 190
Ser Pro Ser Thr Lys Trp His Asn Ser Tyr Arg Glu Pro Phe Glu Gln
195 200 205
His
Claims (15)
1.一种能够结合白细胞介素4(IL-4)受体(IL-4R)的抗体,所述抗体的特征在于,所述抗体包含轻链可变区(VL),所述轻链可变区包含选自下述的CDR1、CDR2和CDR3组合:
(1)SEQ ID NO:1所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:5所示的CDR3;
(2)SEQ ID NO:2所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:5所示的CDR3;
(3)SEQ ID NO:2所示的CDR1,SEQ ID NO:4所示的CDR2和SEQ ID NO:5所示的CDR3;
(4)SEQ ID NO:1所示的CDR1,SEQ ID NO.4所示的CDR2和SEQ ID NO:5所示的CDR3;
(5)SEQ ID NO:2所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:6所示的CDR3;
(6)SEQ ID NO:2所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:7所示的CDR3;
(7)SEQ ID NO:2所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:8所示的CDR3;
(8)SEQ ID NO:1所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:6所示的CDR3;
(9)SEQ ID NO:2所示的CDR1,SEQ ID NO:4所示的CDR2和SEQ ID NO:6所示的CDR3;
(10)SEQ ID NO:2所示的CDR1,SEQ ID NO:4所示的CDR2和SEQ ID NO:8所示的CDR3;
(11)SEQ ID NO:1所示的CDR1,SEQ ID NO:4所示的CDR2和SEQ ID NO:8所示的CDR3;和
(12)SEQ ID NO:1所示的CDR1,SEQ ID NO:3所示的CDR2和SEQ ID NO:8所示的CDR3;
和/或
所述抗体包含重链可变区(VH),所述重链可变区包含选自下述的CDR1、CDR2和CDR3组合:
(1)SEQ ID NO:14所示的CDR1,SEQ ID NO:17所示的CDR2和SEQ ID NO:19所示的CDR3;
(2)SEQ ID NO:14所示的CDR1,SEQ ID NO:18所示的CDR2和SEQ ID NO:19所示的CDR3;
(3)SEQ ID NO:14所示的CDR1,SEQ ID NO:17所示的CDR2和SEQ ID NO:20所示的CDR3;
(4)SEQ ID NO:14所示的CDR1,SEQ ID NO:18所示的CDR2和SEQ ID NO:20所示的CDR3;
(5)SEQ ID NO:15所示的CDR1,SEQ ID NO:17所示的CDR2和SEQ ID NO:19所示的CDR3;
(6)SEQ ID NO:16所示的CDR1,SEQ ID NO:17所示的CDR2和SEQ ID NO:19所示的CDR3;和
(7)SEQ ID NO:14所示的CDR1,SEQ ID NO:18所示的CDR2和SEQ ID NO:19所示的CDR3。
2.根据权利要求1所述的抗体,其特征在于,所述抗体的轻链可变区包含选自下述的FR1、FR2、FR3和FR4组合:
(1)SEQ ID NO:9所示的FR1,SEQ ID NO:10所示的FR2,SEQ ID NO:12所示的FR3和SEQID NO:13所示的FR4;和
(2)SEQ ID NO:9所示的FR1,SEQ ID NO:11所示的FR2,SEQ ID NO:12所示的FR3和SEQID NO:13所示的FR4;
和/或
所述抗体的重链可变区包含选自下述的FR1、FR2、FR3和FR4组合:
(1)SEQ ID NO:21所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQID NO:38所示的FR4;
(2)SEQ ID NO:22所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQID NO:38所示的FR4;
(3)SEQ ID NO:23所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQID NO:38所示的FR4;
(4)SEQ ID NO:24所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQID NO:38所示的FR4;
(5)SEQ ID NO:24所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQID NO:38所示的FR4;
(6)SEQ ID NO:25所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQID NO:38所示的FR4;
(7)SEQ ID NO:26所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQID NO:38所示的FR4;
(8)SEQ ID NO:27所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQID NO:38所示的FR4;
(9)SEQ ID NO:29所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQID NO:38所示的FR4;
(10)SEQ ID NO:30所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(11)SEQ ID NO:24所示的FR1,SEQ ID NO:33所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(12)SEQ ID NO:24所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:36所示的FR3和SEQ ID NO:38所示的FR4;
(13)SEQ ID NO:24所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:37所示的FR3和SEQ ID NO:38所示的FR4;
(14)SEQ ID NO:31所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(15)SEQ ID NO:27所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(16)SEQ ID NO:26所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(17)SEQ ID NO:25所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(18)SEQ ID NO:28所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(19)SEQ ID NO:28所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:34所示的FR3和SEQ ID NO:38所示的FR4;
(20)SEQ ID NO:23所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;
(21)SEQ ID NO:22所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4;和
(22)SEQ ID NO:21所示的FR1,SEQ ID NO:32所示的FR2,SEQ ID NO:35所示的FR3和SEQ ID NO:38所示的FR4。
3.根据权利要求1或2所述的抗体,其特征在于,所述抗体包含选自下述序列所示氨基酸序列的轻链可变区:
SEQ ID NO:39、SEQ ID NO:40、SEQ ID NO:41、SEQ ID NO:42、SEQ ID NO:43、SEQ IDNO:44、SEQ ID NO:45、SEQ ID NO:46、SEQ ID NO:47、SEQ ID NO:48、SEQ ID NO:49、SEQ IDNO:50、SEQ ID NO:51、SEQ ID NO:52、SEQ ID NO:53、SEQ ID NO:54、SEQ ID NO:55、SEQ IDNO:56、SEQ ID NO:57和SEQ ID NO:58;
和/或
所述抗体包含选自下述序列所示氨基酸序列的重链可变区:
SEQ ID NO:59、SEQ ID NO:60、SEQ ID NO:61、SEQ ID NO:62、SEQ ID NO:63、SEQ IDNO:64、SEQ ID NO:65、SEQ ID NO:66、SEQ ID NO:67、SEQ ID NO:68、SEQ ID NO:69、SEQ IDNO:70、SEQ ID NO:71、SEQ ID NO:72、SEQ ID NO:73、SEQ ID NO:74、SEQ ID NO:75、SEQ IDNO:76、SEQ ID NO:77、SEQ ID NO:78、SEQ ID NO:79、SEQ ID NO:80、SEQ ID NO:81、SEQ IDNO:82、SEQ ID NO:83、SEQ ID NO:84、SEQ ID NO:85、SEQ ID NO:86、SEQ ID NO:87、SEQ IDNO:88、SEQ ID NO:89、、SEQ ID NO:90、SEQ ID NO:91、SEQ ID NO:92和SEQ ID NO:93;
优选地,所述抗体包含选自下述的轻链可变区和重链可变区组合:
(1)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.62所示的重链可变区;
(2)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.63所示的重链可变区;
(3)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.59所示的重链可变区;
(4)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.60所示的重链可变区;
(5)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.61所示的重链可变区;
(6)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.67所示的重链可变区;
(7)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.65所示的重链可变区;
(8)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.66所示的重链可变区;
(9)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.64所示的重链可变区;
(10)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.68所示的重链可变区;
(11)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.69所示的重链可变区;
(12)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.70所示的重链可变区;
(13)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.71所示的重链可变区;
(14)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.72所示的重链可变区;
(15)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.73所示的重链可变区;
(16)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.89所示的重链可变区;
(17)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.88所示的重链可变区;
(18)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.87所示的重链可变区;
(19)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.86所示的重链可变区;
(20)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.83所示的重链可变区;
(21)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.82所示的重链可变区;
(22)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.81所示的重链可变区;
(23)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.85所示的重链可变区;
(24)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.84所示的重链可变区;
(25)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(26)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.90所示的重链可变区;
(27)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.74所示的重链可变区;
(28)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.75所示的重链可变区;
(29)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.76所示的重链可变区;
(30)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.77所示的重链可变区;
(31)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.78所示的重链可变区;
(32)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.80所示的重链可变区;
(33)SEQ ID NO.39所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(34)SEQ ID NO.40所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(35)SEQ ID NO.41所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(36)SEQ ID NO.42所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(37)SEQ ID NO.43所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(38)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(39)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.62所示的重链可变区;
(40)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.68所示的重链可变区;
(41)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.72所示的重链可变区;
(42)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.82所示的重链可变区;
(43)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.85所示的重链可变区;
(44)SEQ ID NO.44所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(45)SEQ ID NO.48所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(46)SEQ ID NO.49所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(47)SEQ ID NO.50所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(48)SEQ ID NO.45所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(49)SEQ ID NO.46所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(50)SEQ ID NO.47所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(51)SEQ ID NO.56所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(52)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(53)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.62所示的重链可变区;
(54)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.68所示的重链可变区;
(55)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.72所示的重链可变区;
(56)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.82所示的重链可变区;
(57)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.85所示的重链可变区;
(58)SEQ ID NO.55所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(59)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(60)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.62所示的重链可变区;
(61)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.68所示的重链可变区;
(62)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.72所示的重链可变区;
(63)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.82所示的重链可变区;
(64)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.85所示的重链可变区;
(65)SEQ ID NO.54所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(66)SEQ ID NO.53所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(67)SEQ ID NO.51所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(68)SEQ ID NO.52所示的轻链可变区和SEQ ID NO.92所示的重链可变区;
(69)SEQ ID NO.52所示的轻链可变区和SEQ ID NO.62所示的重链可变区;
(70)SEQ ID NO.52所示的轻链可变区和SEQ ID NO.91所示的重链可变区;
(71)SEQ ID NO.58所示的轻链可变区和SEQ ID NO.92所示的重链可变区;和
(72)SEQ ID NO.57所示的轻链可变区和SEQ ID NO.93所示的重链可变区。
4.根据权利要求1至3中任一项所述的抗体,其特征在于,所述抗体能够结合白细胞介素4(IL-4)受体(IL-4R);
优选地,所述抗体能够结合IL-4Rα,优选结合哺乳动物类IL-4Rα,优选结合人IL-4Rα,更优选结合人可溶性IL-4Rα;
更优选地,所述抗体能够以小于100nM、小于10nM、小于1nM、小于0.5nM或甚至小于0.1nM的亲和力结合IL-4Rα。
5.根据权利要求1至4中任一项所述的抗体,其特征在于,所述抗体为单克隆抗体、完全或部分人源化抗体或嵌合抗体;
优选地,所述抗体为免疫球蛋白,优选为IgA、IgD、IgE、IgG或IgM,更优选为IgG1、IgG2、IgG3或IgG4亚型。
6.一种融合蛋白或偶联物,所述融合蛋白或偶联物包含权利要求1至5中任一项所述的抗体。
7.一种核酸序列,所述核酸序列编码权利要求1至5中任一项所述的抗体的重链可变区和/或轻链可变区;
优选地,所述核酸序列编码权利要求1至5中任一项所述的抗体的重链和/或轻链。
8.一种包含权利要求7所述的核酸序列的载体。
9.一种采用权利要求7所述的核酸序列或权利要求8所述的载体转化或转染的宿主细胞。
10.一种用于生产权利要求1至5中任一项所述的抗体的方法,所述方法包括:由权利要求9所述的核酸序列获得所述抗体的重链可变区和/或轻链可变区,或者获得所述抗体的重链和/或轻链,并与所述抗体的任选其他结构域组装成抗体;或者
所述方法包括:在允许权利要求11所述的宿主细胞表达所述抗体的重链可变区和/或轻链可变区或者所述抗体的重链和/或轻链以组装成所述抗体的情况下,培养所述宿主细胞;
任选地,所述方法还包括回收产生的抗体。
11.一种药物组合物,所述药物组合物包含权利要求1至5中任一项所述的抗体、权利要求6所述的融合蛋白或偶联物、权利要求7所述的核酸序列、权利要求8所述的载体、权利要求9所述的宿主细胞和/或通过权利要求10所述的方法生产的抗体;
优选地,所述药物组合物为药物制剂,所述药物制剂优选为注射剂剂型;
优选地,所述药物组合物或药物制剂还包含药学上可接受的载体或赋形剂;
优选地,所述药物组合物或药物制剂还包含至少一种选自下述的药物:平喘药如沙丁胺醇等,抗组胺药如氯雷他定等,免疫抑制剂如他克莫司、吡美莫司等,M受体阻断剂如异丙托溴铵等,白三烯受体阻断药如孟鲁司特等,磷酸二酯酶抑制剂如茶碱等,非甾体抗炎药如5-氨基水杨酸等,激素类如倍氯米松,布地奈德等。
12.权利要求1至5中任一项所述的抗体、权利要求6所述的融合蛋白或偶联物、权利要求7所述的核酸序列、权利要求8所述的载体、权利要求9所述的宿主细胞和/或通过权利要求10所述的方法生产的抗体在制备用于预防、治疗或改善炎症或过敏性疾病的药物中的用途;
优选地,所述炎症或过敏性疾病包括自身免疫学疾病,例如过敏性皮炎、哮喘、嗜酸性粒细胞食管炎、湿疹、过敏性鼻炎、鼻息肉、类风湿性关节炎等。
13.一种预防、治疗或改善受试者的炎症或过敏性疾病的方法,所述方法包括给有此需要的受试者施用权利要求1至5中任一项所述的抗体、权利要求6所述的融合蛋白或偶联物、权利要求7所述的核酸序列、权利要求8所述的载体、权利要求9所述的宿主细胞和/或通过权利要求10所述的方法生产的抗体;
优选地,所述受试者为哺乳类动物;
更优选地,所述受试者为人;
优选地,所述炎症或过敏性疾病包括自身免疫学疾病,例如过敏性皮炎、哮喘、嗜酸性粒细胞食管炎、湿疹、过敏性鼻炎、鼻息肉、类风湿性关节炎等。
14.根据权利要求13所述的方法,其特征在于,所述方法还包括给受试者施用至少一种选自下述的药物:平喘药如沙丁胺醇等,抗组胺药如氯雷他定等,免疫抑制剂如他克莫司、吡美莫司等,M受体阻断剂如异丙托溴铵等,白三烯受体阻断药如孟鲁司特等,磷酸二酯酶抑制剂如茶碱等,非甾体抗炎药如5-氨基水杨酸等,激素类如倍氯米松、布地奈德等;
优选地,所述药物与权利要求1至5中任一项所述的抗体、权利要求6所述的融合蛋白或偶联物、权利要求7所述的核酸序列、权利要求8所述的载体、权利要求9所述的宿主细胞和/或通过权利要求10所述的方法生产的抗体同时或顺序施用。
15.一种试剂盒,所述试剂盒包括权利要求1至5中任一项所述的抗体、权利要求6所述的融合蛋白或偶联物、权利要求7所述的核酸序列、权利要求8所述的载体、权利要求9所述的宿主细胞和/或通过权利要求10所述的方法生产的抗体。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110760590.8A CN113372446A (zh) | 2016-06-08 | 2016-06-08 | 用于结合白细胞介素4受体的抗体 |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201610399254.4A CN107474134B (zh) | 2016-06-08 | 2016-06-08 | 用于结合白细胞介素4受体的抗体 |
CN202110760590.8A CN113372446A (zh) | 2016-06-08 | 2016-06-08 | 用于结合白细胞介素4受体的抗体 |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610399254.4A Division CN107474134B (zh) | 2016-06-08 | 2016-06-08 | 用于结合白细胞介素4受体的抗体 |
Publications (1)
Publication Number | Publication Date |
---|---|
CN113372446A true CN113372446A (zh) | 2021-09-10 |
Family
ID=60577597
Family Applications (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110760590.8A Withdrawn CN113372446A (zh) | 2016-06-08 | 2016-06-08 | 用于结合白细胞介素4受体的抗体 |
CN201610399254.4A Active CN107474134B (zh) | 2016-06-08 | 2016-06-08 | 用于结合白细胞介素4受体的抗体 |
Family Applications After (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201610399254.4A Active CN107474134B (zh) | 2016-06-08 | 2016-06-08 | 用于结合白细胞介素4受体的抗体 |
Country Status (16)
Country | Link |
---|---|
US (3) | US20190177408A1 (zh) |
EP (1) | EP3470430A4 (zh) |
JP (2) | JP7025356B2 (zh) |
KR (2) | KR102502988B1 (zh) |
CN (2) | CN113372446A (zh) |
AU (1) | AU2017276473A1 (zh) |
BR (1) | BR112018074325A2 (zh) |
CA (1) | CA3026568A1 (zh) |
IL (1) | IL263268B2 (zh) |
MX (1) | MX2018014941A (zh) |
MY (1) | MY189035A (zh) |
PH (1) | PH12018502544A1 (zh) |
SA (1) | SA518400605B1 (zh) |
SG (1) | SG11201810855VA (zh) |
WO (1) | WO2017211319A1 (zh) |
ZA (1) | ZA201808209B (zh) |
Families Citing this family (25)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113372446A (zh) | 2016-06-08 | 2021-09-10 | 苏州康乃德生物医药有限公司 | 用于结合白细胞介素4受体的抗体 |
KR102652133B1 (ko) * | 2018-02-01 | 2024-03-29 | 베이징 카윈 테크놀로지 쉐어-홀딩 컴퍼니 리미티드 | IL-4Rα 항체 및 그 용도 |
CN108373505B (zh) * | 2018-04-20 | 2019-08-20 | 北京智仁美博生物科技有限公司 | 抗il-4r抗体及其用途 |
MX2020012064A (es) | 2018-05-13 | 2021-04-13 | Regeneron Pharma | Metodos para tratar la dermatitis atopica administrando un inhibidor de il-4r. |
CN110540590B (zh) * | 2018-05-29 | 2023-08-18 | 康诺亚生物医药科技(成都)有限公司 | 一种自免疫抑制物的开发和应用 |
CN110872349A (zh) | 2018-09-04 | 2020-03-10 | 三生国健药业(上海)股份有限公司 | 结合人il-4r的抗体、其制备方法和用途 |
CN111518211B (zh) | 2018-12-25 | 2022-06-21 | 江苏荃信生物医药股份有限公司 | 抗人白介素4受体α单克隆抗体的制药用途 |
CN111686247B (zh) * | 2019-03-13 | 2022-07-29 | 苏州康乃德生物医药有限公司 | 包含人白介素-4受体α的抗体的液体组合物 |
CN112010977B (zh) * | 2019-05-29 | 2022-04-26 | 山东博安生物技术股份有限公司 | 抗白介素4受体(il-4r)的抗体及其应用 |
CN115427450A (zh) | 2020-03-27 | 2022-12-02 | 瑞泽恩制药公司 | 通过施用il-4r拮抗剂治疗特应性皮炎的方法 |
CN113527485A (zh) | 2020-04-17 | 2021-10-22 | 上海麦济生物技术有限公司 | 抗人白细胞介素-4受体α抗体及其制备方法和应用 |
BR112022022235A2 (pt) | 2020-05-22 | 2023-03-28 | Regeneron Pharma | Métodos para tratamento de esofagite eosinofílica através da administração de inibidor de il-4r |
CN114957472B (zh) * | 2020-06-22 | 2023-10-31 | 南京融捷康生物科技有限公司 | 抗IL-4Rα的单域抗体以及应用和药物 |
CN114555639B (zh) * | 2020-09-10 | 2023-12-12 | 舒泰神(北京)生物制药股份有限公司 | 特异性识别白细胞介素-4受体α的抗体及其用途 |
MX2023003942A (es) | 2020-10-05 | 2023-06-02 | Sanofi Biotechnology | Metodos para el tratamiento del asma en sujetos pediatricos mediante la administracion de un antagonista del il-4r. |
EP4274609A1 (en) | 2021-01-08 | 2023-11-15 | Regeneron Pharmaceuticals, Inc. | Methods for treating peanut allergy and enhancing peanut allergen-specific immunotherapy by administering an il-4r antagonist |
AU2022316939A1 (en) | 2021-07-26 | 2024-03-07 | Regeneron Pharmaceuticals, Inc. | Methods for treating chronic spontaneous urticaria by administering an il-4r antagonist |
US20230167171A1 (en) | 2021-08-23 | 2023-06-01 | Regeneron Pharmaceuticals, Inc. | Methods for treating atopic dermatitis by administering an il-4r antagonist |
WO2023069976A1 (en) | 2021-10-20 | 2023-04-27 | Sanofi Biotechnology | Methods for treating prurigo nodularis by administering an il-4r antagonist |
WO2023130010A1 (en) | 2021-12-30 | 2023-07-06 | Regeneron Pharmaceuticals, Inc. | Methods for attenuating atopic march by administering an il-4/il-13 antagonist |
WO2023167871A1 (en) | 2022-03-02 | 2023-09-07 | Regeneron Pharmaceuticals, Inc. | Manufacturing process for high titer antibody |
US20240024472A1 (en) | 2022-05-02 | 2024-01-25 | Regeneron Pharmaceuticals, Inc. | Anti-Interleukin-4 Receptor (IL-4R) Antibody Formulations |
WO2023215750A2 (en) | 2022-05-02 | 2023-11-09 | Regeneron Pharmaceuticals, Inc. | Methods for reducing lipase activity |
WO2024011251A1 (en) | 2022-07-08 | 2024-01-11 | Regeneron Pharmaceuticals, Inc. | Methods for treating eosinophilic esophagitis in pediatric by administering an il-4r antagonist |
WO2024047021A1 (en) | 2022-08-29 | 2024-03-07 | Sanofi | Methods for treating chronic inducible cold urticaria by administering an il-4r antagonist |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1886426A (zh) * | 2003-11-07 | 2006-12-27 | 伊姆尼斯公司 | 结合白细胞介素-4受体的抗体 |
US7186809B2 (en) * | 2000-05-26 | 2007-03-06 | Immunex Corporation | Methods and compositions relating to anti-interleukin-4 receptor antibodies |
CN103998053A (zh) * | 2011-12-13 | 2014-08-20 | 皮里斯股份公司 | 通过抑制il-4和/或il-13与其各自的受体结合来预防或治疗某些障碍的方法 |
Family Cites Families (24)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2007534305A (ja) * | 2003-11-07 | 2007-11-29 | アムジェン インコーポレイテッド | サル免疫グロブリン配列 |
MY149079A (en) | 2006-10-02 | 2013-07-15 | Regeneron Pharma | High affinity human antibodies to human il-4 receptor |
US7608693B2 (en) * | 2006-10-02 | 2009-10-27 | Regeneron Pharmaceuticals, Inc. | High affinity human antibodies to human IL-4 receptor |
US8092804B2 (en) * | 2007-12-21 | 2012-01-10 | Medimmune Limited | Binding members for interleukin-4 receptor alpha (IL-4Rα)-173 |
WO2009121847A2 (en) * | 2008-04-02 | 2009-10-08 | Apogenix Gmbh | Binding agents directed against il-4 receptor for the treatment of tumors, inflammatory and immunological disorders |
PL2624865T3 (pl) | 2010-10-06 | 2018-11-30 | Regeneron Pharmaceuticals, Inc. | Stabilizowane preparaty zawierające przeciwciała przeciwko receptorowi interleukiny-4 (IL-4R) |
HUE038305T2 (hu) * | 2010-11-17 | 2018-10-29 | Chugai Pharmaceutical Co Ltd | A VIII vérkoagulációs faktor mûködésére alternatív funkciójú multispecifikus antigén-kötõ molekula |
JO3756B1 (ar) * | 2010-11-23 | 2021-01-31 | Regeneron Pharma | اجسام مضادة بشرية لمستقبلات الجلوكاجون |
US8790651B2 (en) * | 2011-07-21 | 2014-07-29 | Zoetis Llc | Interleukin-31 monoclonal antibody |
AR092177A1 (es) | 2012-08-21 | 2015-03-25 | Sanofi Sa | Metodos para tratar o prevenir asma administrando un antagonista de il-4r |
ES2675779T3 (es) | 2012-09-07 | 2018-07-12 | Regeneron Pharmaceuticals, Inc. | Métodos para el tratamiento de la dermatitis atópica administrando un antagonista de IL-4R |
TWI634900B (zh) | 2013-07-11 | 2018-09-11 | 再生元醫藥公司 | 藉由投與il-4r抑制劑治療嗜酸性食道炎的方法 |
RU2016134838A (ru) | 2014-01-27 | 2018-03-07 | МЕДИММЬЮН, ЭлЭлСи | Дипептидил пептидаза-4 (dpp4/cd26) как периферический биомаркер активации il-13 в астматическом легком |
MX2016011132A (es) | 2014-02-28 | 2016-12-08 | Regeneron Pharma | Métodos para tratar una infección cutánea mediante la administración de un antagonista de il-4r. |
CN113372446A (zh) | 2016-06-08 | 2021-09-10 | 苏州康乃德生物医药有限公司 | 用于结合白细胞介素4受体的抗体 |
KR102462039B1 (ko) | 2016-09-01 | 2022-11-02 | 리제너론 파아마슈티컬스, 인크. | Il-4r 길항제를 투여함에 의해 알레르기를 예방하거나 치료하기 위한 방법 |
WO2018057776A1 (en) | 2016-09-22 | 2018-03-29 | Regeneron Pharmaceuticals, Inc. | Methods for treating severe atopic dermatitis by administering an il-4r inhibitor |
CN108339118A (zh) | 2017-01-23 | 2018-07-31 | 瑞阳(苏州)生物科技有限公司 | 治疗或预防阻塞性睡眠呼吸暂停的药物组合物 |
US11053309B2 (en) | 2017-08-04 | 2021-07-06 | Regeneron Pharmaceuticals, Inc. | Methods for treating active eosinophilic esophagitis |
MX2020004467A (es) | 2017-10-30 | 2020-10-22 | Sanofi Biotechnology | Metodos para el tratamiento o la prevencion del asma mediante la administracion de antagonista de il-4r. |
MX2020012064A (es) | 2018-05-13 | 2021-04-13 | Regeneron Pharma | Metodos para tratar la dermatitis atopica administrando un inhibidor de il-4r. |
CN113166259A (zh) | 2018-11-09 | 2021-07-23 | 亚洲大学校产学协力团 | 对人IL-4受体α具有高亲和力的人抗体及其用途 |
CN113677707A (zh) | 2019-03-06 | 2021-11-19 | 瑞泽恩制药公司 | 用于在治疗癌症中增强效力的il-4/il-13途径抑制剂 |
CN111686247B (zh) | 2019-03-13 | 2022-07-29 | 苏州康乃德生物医药有限公司 | 包含人白介素-4受体α的抗体的液体组合物 |
-
2016
- 2016-06-08 CN CN202110760590.8A patent/CN113372446A/zh not_active Withdrawn
- 2016-06-08 CN CN201610399254.4A patent/CN107474134B/zh active Active
-
2017
- 2017-06-08 CA CA3026568A patent/CA3026568A1/en active Pending
- 2017-06-08 MY MYPI2018002201A patent/MY189035A/en unknown
- 2017-06-08 AU AU2017276473A patent/AU2017276473A1/en active Pending
- 2017-06-08 KR KR1020227022353A patent/KR102502988B1/ko active IP Right Grant
- 2017-06-08 WO PCT/CN2017/087592 patent/WO2017211319A1/zh unknown
- 2017-06-08 KR KR1020197000454A patent/KR102417217B1/ko active IP Right Grant
- 2017-06-08 JP JP2018564202A patent/JP7025356B2/ja active Active
- 2017-06-08 SG SG11201810855VA patent/SG11201810855VA/en unknown
- 2017-06-08 MX MX2018014941A patent/MX2018014941A/es unknown
- 2017-06-08 BR BR112018074325A patent/BR112018074325A2/pt unknown
- 2017-06-08 US US16/307,930 patent/US20190177408A1/en not_active Abandoned
- 2017-06-08 IL IL263268A patent/IL263268B2/en unknown
- 2017-06-08 EP EP17809758.0A patent/EP3470430A4/en active Pending
-
2018
- 2018-12-03 PH PH12018502544A patent/PH12018502544A1/en unknown
- 2018-12-05 ZA ZA2018/08209A patent/ZA201808209B/en unknown
- 2018-12-06 SA SA518400605A patent/SA518400605B1/ar unknown
-
2020
- 2020-03-04 US US16/809,411 patent/US10774141B2/en active Active
- 2020-08-14 US US16/994,464 patent/US11866491B2/en active Active
-
2022
- 2022-02-10 JP JP2022019435A patent/JP2022065063A/ja active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US7186809B2 (en) * | 2000-05-26 | 2007-03-06 | Immunex Corporation | Methods and compositions relating to anti-interleukin-4 receptor antibodies |
CN1886426A (zh) * | 2003-11-07 | 2006-12-27 | 伊姆尼斯公司 | 结合白细胞介素-4受体的抗体 |
CN103998053A (zh) * | 2011-12-13 | 2014-08-20 | 皮里斯股份公司 | 通过抑制il-4和/或il-13与其各自的受体结合来预防或治疗某些障碍的方法 |
Also Published As
Publication number | Publication date |
---|---|
CN107474134B (zh) | 2021-07-27 |
EP3470430A1 (en) | 2019-04-17 |
MX2018014941A (es) | 2019-03-07 |
JP7025356B2 (ja) | 2022-03-03 |
SA518400605B1 (ar) | 2022-10-30 |
US20210079088A1 (en) | 2021-03-18 |
EP3470430A4 (en) | 2020-06-03 |
US11866491B2 (en) | 2024-01-09 |
MY189035A (en) | 2022-01-20 |
IL263268A (en) | 2018-12-31 |
KR20220093409A (ko) | 2022-07-05 |
PH12018502544A1 (en) | 2019-10-07 |
CN107474134A (zh) | 2017-12-15 |
US20190177408A1 (en) | 2019-06-13 |
ZA201808209B (en) | 2019-09-25 |
IL263268B1 (en) | 2023-06-01 |
KR102417217B1 (ko) | 2022-07-05 |
JP2019520816A (ja) | 2019-07-25 |
BR112018074325A2 (pt) | 2019-10-01 |
CA3026568A1 (en) | 2017-12-14 |
KR20190015757A (ko) | 2019-02-14 |
IL263268B2 (en) | 2023-10-01 |
RU2019100002A3 (zh) | 2020-10-07 |
RU2019100002A (ru) | 2020-07-14 |
KR102502988B1 (ko) | 2023-02-23 |
US10774141B2 (en) | 2020-09-15 |
US20200255514A1 (en) | 2020-08-13 |
SG11201810855VA (en) | 2019-01-30 |
AU2017276473A1 (en) | 2019-01-03 |
JP2022065063A (ja) | 2022-04-26 |
WO2017211319A1 (zh) | 2017-12-14 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN107474134B (zh) | 用于结合白细胞介素4受体的抗体 | |
WO2017215524A1 (zh) | 抗人白细胞介素-17a单克隆抗体、其制备方法和应用 | |
CN111393526B (zh) | 抗gdf15中和性单克隆抗体及其应用 | |
EP4289861A1 (en) | Antibodies against human tslp and use thereof | |
CN111040035B (zh) | 针对il-17ra蛋白的抗体及其制备方法和应用 | |
CN107074954A (zh) | Il‑6r相关疾病的治疗 | |
CN115461368A (zh) | 抗人白细胞介素-4受体α抗体及其制备方法和应用 | |
WO2022143611A1 (zh) | 靶向bcma的单域抗体 | |
EP4242229A1 (en) | Antibody against human thymic stromal lymphopoietin, preparation method therefor and application thereof | |
RU2774446C2 (ru) | Антитело для специфического связывания с рецептором интерлейкина 4 | |
WO2022117079A1 (zh) | 结合胸腺基质淋巴细胞生成素的抗体及其应用 | |
CN111234021B (zh) | 一种抗ccr5抗体及其在治疗肿瘤中的应用 | |
WO2022100664A1 (zh) | 一种tslp抗原结合蛋白及其应用 | |
WO2022116858A1 (zh) | 一种抗tslp抗体药物组合物及其用途 | |
RU2617537C1 (ru) | Система для получения аутологичной сыворотки | |
CN116284401A (zh) | 人源抗il-1r3抗体及其应用 | |
TW202321300A (zh) | Il-11人源化抗體及其應用 | |
CN117843777A (zh) | 用于炎性疾病治疗的新型纳米抗体及其产品和方法 | |
CN116102659A (zh) | 一种抗il-17/vegf双功能融合蛋白及其用途 | |
CN117843778A (zh) | 针对白介素家族成员的新型纳米抗体及其产品和应用 | |
CN109705217A (zh) | 抗il-13抗体及其用途 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
WW01 | Invention patent application withdrawn after publication |
Application publication date: 20210910 |
|
WW01 | Invention patent application withdrawn after publication |