CN114395040A - 再生蛋白reg1a单克隆抗体及其应用 - Google Patents
再生蛋白reg1a单克隆抗体及其应用 Download PDFInfo
- Publication number
- CN114395040A CN114395040A CN202210122652.7A CN202210122652A CN114395040A CN 114395040 A CN114395040 A CN 114395040A CN 202210122652 A CN202210122652 A CN 202210122652A CN 114395040 A CN114395040 A CN 114395040A
- Authority
- CN
- China
- Prior art keywords
- light chain
- heavy chain
- amino acid
- acid sequence
- reg1a
- 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.)
- Granted
Links
- 102100027361 Lithostathine-1-alpha Human genes 0.000 title claims abstract description 70
- 101000581802 Homo sapiens Lithostathine-1-alpha Proteins 0.000 title claims abstract description 67
- 150000007523 nucleic acids Chemical class 0.000 claims description 13
- 108090000623 proteins and genes Proteins 0.000 claims description 13
- 108020004707 nucleic acids Proteins 0.000 claims description 11
- 102000039446 nucleic acids Human genes 0.000 claims description 11
- 230000008929 regeneration Effects 0.000 claims description 11
- 238000011069 regeneration method Methods 0.000 claims description 11
- 238000002360 preparation method Methods 0.000 claims description 8
- 102000004169 proteins and genes Human genes 0.000 claims description 8
- 108010014691 Lithostathine Proteins 0.000 claims description 7
- 238000001514 detection method Methods 0.000 claims description 4
- 230000002285 radioactive effect Effects 0.000 claims description 3
- 230000002255 enzymatic effect Effects 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 20
- 238000009007 Diagnostic Kit Methods 0.000 claims 1
- 102100023882 Endoribonuclease ZC3H12A Human genes 0.000 claims 1
- 101710112715 Endoribonuclease ZC3H12A Proteins 0.000 claims 1
- 238000011161 development Methods 0.000 abstract description 6
- 201000010099 disease Diseases 0.000 abstract description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 5
- 238000003745 diagnosis Methods 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 27
- 150000001413 amino acids Chemical class 0.000 description 23
- 241000699666 Mus <mouse, genus> Species 0.000 description 14
- 210000004408 hybridoma Anatomy 0.000 description 12
- 230000003053 immunization Effects 0.000 description 9
- 238000002649 immunization Methods 0.000 description 9
- 239000002609 medium Substances 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- 206010035226 Plasma cell myeloma Diseases 0.000 description 8
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 8
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 8
- 201000000050 myeloid neoplasm Diseases 0.000 description 8
- 239000002244 precipitate Substances 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 239000013612 plasmid Substances 0.000 description 6
- 210000004989 spleen cell Anatomy 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000001262 western blot Methods 0.000 description 6
- 241000699670 Mus sp. Species 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 239000013598 vector Substances 0.000 description 5
- 206010003445 Ascites Diseases 0.000 description 4
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 4
- 239000000427 antigen Substances 0.000 description 4
- 102000036639 antigens Human genes 0.000 description 4
- 108091007433 antigens Proteins 0.000 description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 210000004988 splenocyte Anatomy 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- 108010073969 valyllysine Proteins 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 3
- 102000016997 Lithostathine Human genes 0.000 description 3
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 3
- 238000012408 PCR amplification Methods 0.000 description 3
- 210000000683 abdominal cavity Anatomy 0.000 description 3
- 239000002671 adjuvant Substances 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 230000001580 bacterial effect Effects 0.000 description 3
- 230000007910 cell fusion Effects 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 206010012601 diabetes mellitus Diseases 0.000 description 3
- 238000010586 diagram Methods 0.000 description 3
- 230000002183 duodenal effect Effects 0.000 description 3
- 102000037865 fusion proteins Human genes 0.000 description 3
- 108020001507 fusion proteins Proteins 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 238000000746 purification Methods 0.000 description 3
- 238000012216 screening Methods 0.000 description 3
- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 206010061218 Inflammation Diseases 0.000 description 2
- 241001529936 Murinae Species 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 2
- 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 2
- 238000001042 affinity chromatography Methods 0.000 description 2
- 229960000723 ampicillin Drugs 0.000 description 2
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 2
- 230000004054 inflammatory process Effects 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 210000004153 islets of langerhan Anatomy 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 210000005084 renal tissue Anatomy 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- 239000001993 wax Substances 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- HOVPGJUNRLMIOZ-CIUDSAMLSA-N Ala-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C)N HOVPGJUNRLMIOZ-CIUDSAMLSA-N 0.000 description 1
- ARHJJAAWNWOACN-FXQIFTODSA-N Ala-Ser-Val Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O ARHJJAAWNWOACN-FXQIFTODSA-N 0.000 description 1
- CREYEAPXISDKSB-FQPOAREZSA-N Ala-Thr-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CREYEAPXISDKSB-FQPOAREZSA-N 0.000 description 1
- ZATRYQNPUHGXCU-DTWKUNHWSA-N Arg-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CCCN=C(N)N)N)C(=O)O ZATRYQNPUHGXCU-DTWKUNHWSA-N 0.000 description 1
- AOHKLEBWKMKITA-IHRRRGAJSA-N Arg-Phe-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N AOHKLEBWKMKITA-IHRRRGAJSA-N 0.000 description 1
- KLKHFFMNGWULBN-VKHMYHEASA-N Asn-Gly Chemical compound NC(=O)C[C@H](N)C(=O)NCC(O)=O KLKHFFMNGWULBN-VKHMYHEASA-N 0.000 description 1
- BXUHCIXDSWRSBS-CIUDSAMLSA-N Asn-Leu-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O BXUHCIXDSWRSBS-CIUDSAMLSA-N 0.000 description 1
- XYBJLTKSGFBLCS-QXEWZRGKSA-N Asp-Arg-Val Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)CC(O)=O XYBJLTKSGFBLCS-QXEWZRGKSA-N 0.000 description 1
- QNFRBNZGVVKBNJ-PEFMBERDSA-N Asp-Ile-Gln Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)O)N QNFRBNZGVVKBNJ-PEFMBERDSA-N 0.000 description 1
- WMLFFCRUSPNENW-ZLUOBGJFSA-N Asp-Ser-Ala Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O WMLFFCRUSPNENW-ZLUOBGJFSA-N 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 238000011537 Coomassie blue staining Methods 0.000 description 1
- AMRLSQGGERHDHJ-FXQIFTODSA-N Cys-Ala-Arg Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O AMRLSQGGERHDHJ-FXQIFTODSA-N 0.000 description 1
- 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 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000672609 Escherichia coli BL21 Species 0.000 description 1
- LVSYIKGMLRHKME-IUCAKERBSA-N Gln-Gly-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CCC(=O)N)N LVSYIKGMLRHKME-IUCAKERBSA-N 0.000 description 1
- KVQOVQVGVKDZNW-GUBZILKMSA-N Gln-Ser-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)N)N KVQOVQVGVKDZNW-GUBZILKMSA-N 0.000 description 1
- IESFZVCAVACGPH-PEFMBERDSA-N Glu-Asp-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CCC(O)=O IESFZVCAVACGPH-PEFMBERDSA-N 0.000 description 1
- GJBUAAAIZSRCDC-GVXVVHGQSA-N Glu-Leu-Val Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(O)=O GJBUAAAIZSRCDC-GVXVVHGQSA-N 0.000 description 1
- GRIRDMVMJJDZKV-RCOVLWMOSA-N Gly-Asn-Val Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(O)=O GRIRDMVMJJDZKV-RCOVLWMOSA-N 0.000 description 1
- UQJNXZSSGQIPIQ-FBCQKBJTSA-N Gly-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)CN UQJNXZSSGQIPIQ-FBCQKBJTSA-N 0.000 description 1
- NTBOEZICHOSJEE-YUMQZZPRSA-N Gly-Lys-Ser Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O NTBOEZICHOSJEE-YUMQZZPRSA-N 0.000 description 1
- NVTPVQLIZCOJFK-FOHZUACHSA-N Gly-Thr-Asp Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O NVTPVQLIZCOJFK-FOHZUACHSA-N 0.000 description 1
- TVTZEOHWHUVYCG-KYNKHSRBSA-N Gly-Thr-Thr Chemical compound [H]NCC(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O TVTZEOHWHUVYCG-KYNKHSRBSA-N 0.000 description 1
- CUVBTVWFVIIDOC-YEPSODPASA-N Gly-Thr-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)CN CUVBTVWFVIIDOC-YEPSODPASA-N 0.000 description 1
- GBYYQVBXFVDJPJ-WLTAIBSBSA-N Gly-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)CN)O GBYYQVBXFVDJPJ-WLTAIBSBSA-N 0.000 description 1
- ZHHLTWUOWXHVQJ-YUMQZZPRSA-N His-Ser-Gly Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CO)C(=O)NCC(=O)O)N ZHHLTWUOWXHVQJ-YUMQZZPRSA-N 0.000 description 1
- ZNTSGDNUITWTRA-WDSOQIARSA-N His-Trp-Val Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](C(C)C)C(O)=O ZNTSGDNUITWTRA-WDSOQIARSA-N 0.000 description 1
- 102000018251 Hypoxanthine Phosphoribosyltransferase Human genes 0.000 description 1
- 108010091358 Hypoxanthine Phosphoribosyltransferase Proteins 0.000 description 1
- KIAOPHMUNPPGEN-PEXQALLHSA-N Ile-Gly-His Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N KIAOPHMUNPPGEN-PEXQALLHSA-N 0.000 description 1
- UIEZQYNXCYHMQS-BJDJZHNGSA-N Ile-Lys-Ala Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(=O)O)N UIEZQYNXCYHMQS-BJDJZHNGSA-N 0.000 description 1
- FHPZJWJWTWZKNA-LLLHUVSDSA-N Ile-Phe-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N FHPZJWJWTWZKNA-LLLHUVSDSA-N 0.000 description 1
- JODPUDMBQBIWCK-GHCJXIJMSA-N Ile-Ser-Asn Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O JODPUDMBQBIWCK-GHCJXIJMSA-N 0.000 description 1
- PXKACEXYLPBMAD-JBDRJPRFSA-N Ile-Ser-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PXKACEXYLPBMAD-JBDRJPRFSA-N 0.000 description 1
- TYYLDKGBCJGJGW-UHFFFAOYSA-N L-tryptophan-L-tyrosine Natural products C=1NC2=CC=CC=C2C=1CC(N)C(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 TYYLDKGBCJGJGW-UHFFFAOYSA-N 0.000 description 1
- LZDNBBYBDGBADK-UHFFFAOYSA-N L-valyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C(C)C)C(O)=O)=CNC2=C1 LZDNBBYBDGBADK-UHFFFAOYSA-N 0.000 description 1
- 108090001090 Lectins Proteins 0.000 description 1
- 102000004856 Lectins Human genes 0.000 description 1
- UCOCBWDBHCUPQP-DCAQKATOSA-N Leu-Arg-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O UCOCBWDBHCUPQP-DCAQKATOSA-N 0.000 description 1
- USTCFDAQCLDPBD-XIRDDKMYSA-N Leu-Asn-Trp Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N USTCFDAQCLDPBD-XIRDDKMYSA-N 0.000 description 1
- 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 1
- UWKNTTJNVSYXPC-CIUDSAMLSA-N Lys-Ala-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN UWKNTTJNVSYXPC-CIUDSAMLSA-N 0.000 description 1
- PRSBSVAVOQOAMI-BJDJZHNGSA-N Lys-Ile-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCCCN PRSBSVAVOQOAMI-BJDJZHNGSA-N 0.000 description 1
- SKRGVGLIRUGANF-AVGNSLFASA-N Lys-Leu-Glu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SKRGVGLIRUGANF-AVGNSLFASA-N 0.000 description 1
- AIRZWUMAHCDDHR-KKUMJFAQSA-N Lys-Leu-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O AIRZWUMAHCDDHR-KKUMJFAQSA-N 0.000 description 1
- AZOFEHCPMBRNFD-BZSNNMDCSA-N Lys-Phe-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](CCCCN)C(O)=O)CC1=CC=CC=C1 AZOFEHCPMBRNFD-BZSNNMDCSA-N 0.000 description 1
- WGILOYIKJVQUPT-DCAQKATOSA-N Lys-Pro-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(O)=O WGILOYIKJVQUPT-DCAQKATOSA-N 0.000 description 1
- PDIDTSZKKFEDMB-UWVGGRQHSA-N Lys-Pro-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O PDIDTSZKKFEDMB-UWVGGRQHSA-N 0.000 description 1
- SPSSJSICDYYTQN-HJGDQZAQSA-N Met-Thr-Gln Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCC(N)=O SPSSJSICDYYTQN-HJGDQZAQSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- 108010066427 N-valyltryptophan Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 206010033645 Pancreatitis Diseases 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- FRPVPGRXUKFEQE-YDHLFZDLSA-N Phe-Asp-Val Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O FRPVPGRXUKFEQE-YDHLFZDLSA-N 0.000 description 1
- FSPGBMWPNMRWDB-AVGNSLFASA-N Phe-Cys-Gln Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N FSPGBMWPNMRWDB-AVGNSLFASA-N 0.000 description 1
- RAGOJJCBGXARPO-XVSYOHENSA-N Phe-Thr-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H]([C@H](O)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 RAGOJJCBGXARPO-XVSYOHENSA-N 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 229920002684 Sepharose Polymers 0.000 description 1
- WDXYVIIVDIDOSX-DCAQKATOSA-N Ser-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CCCN=C(N)N WDXYVIIVDIDOSX-DCAQKATOSA-N 0.000 description 1
- ULVMNZOKDBHKKI-ACZMJKKPSA-N Ser-Gln-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O ULVMNZOKDBHKKI-ACZMJKKPSA-N 0.000 description 1
- KDGARKCAKHBEDB-NKWVEPMBSA-N Ser-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CO)N)C(=O)O KDGARKCAKHBEDB-NKWVEPMBSA-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
- YUJLIIRMIAGMCQ-CIUDSAMLSA-N Ser-Leu-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O YUJLIIRMIAGMCQ-CIUDSAMLSA-N 0.000 description 1
- XJDMUQCLVSCRSJ-VZFHVOOUSA-N Ser-Thr-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O XJDMUQCLVSCRSJ-VZFHVOOUSA-N 0.000 description 1
- 239000006180 TBST buffer Substances 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- JQAWYCUUFIMTHE-WLTAIBSBSA-N Thr-Gly-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JQAWYCUUFIMTHE-WLTAIBSBSA-N 0.000 description 1
- NCXVJIQMWSGRHY-KXNHARMFSA-N Thr-Leu-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N)O NCXVJIQMWSGRHY-KXNHARMFSA-N 0.000 description 1
- BIBYEFRASCNLAA-CDMKHQONSA-N Thr-Phe-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CC=CC=C1 BIBYEFRASCNLAA-CDMKHQONSA-N 0.000 description 1
- XHWCDRUPDNSDAZ-XKBZYTNZSA-N Thr-Ser-Glu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N)O XHWCDRUPDNSDAZ-XKBZYTNZSA-N 0.000 description 1
- ZMYCLHFLHRVOEA-HEIBUPTGSA-N Thr-Thr-Ser Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ZMYCLHFLHRVOEA-HEIBUPTGSA-N 0.000 description 1
- FYBFTPLPAXZBOY-KKHAAJSZSA-N Thr-Val-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O FYBFTPLPAXZBOY-KKHAAJSZSA-N 0.000 description 1
- FEZASNVQLJQBHW-CABZTGNLSA-N Trp-Gly-Ala Chemical compound C1=CC=C2C(C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O)=CNC2=C1 FEZASNVQLJQBHW-CABZTGNLSA-N 0.000 description 1
- MBFJIHUHHCJBSN-AVGNSLFASA-N Tyr-Asn-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O MBFJIHUHHCJBSN-AVGNSLFASA-N 0.000 description 1
- ZNFPUOSTMUMUDR-JRQIVUDYSA-N Tyr-Asn-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZNFPUOSTMUMUDR-JRQIVUDYSA-N 0.000 description 1
- QUILOGWWLXMSAT-IHRRRGAJSA-N Tyr-Gln-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O QUILOGWWLXMSAT-IHRRRGAJSA-N 0.000 description 1
- XDGPTBVOSHKDFT-KKUMJFAQSA-N Tyr-Met-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCC(O)=O)C(O)=O XDGPTBVOSHKDFT-KKUMJFAQSA-N 0.000 description 1
- MQGGXGKQSVEQHR-KKUMJFAQSA-N Tyr-Ser-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 MQGGXGKQSVEQHR-KKUMJFAQSA-N 0.000 description 1
- KSGKJSFPWSMJHK-JNPHEJMOSA-N Tyr-Tyr-Thr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KSGKJSFPWSMJHK-JNPHEJMOSA-N 0.000 description 1
- SSYBNWFXCFNRFN-GUBZILKMSA-N Val-Pro-Ser Chemical compound CC(C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SSYBNWFXCFNRFN-GUBZILKMSA-N 0.000 description 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
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 108010086434 alanyl-seryl-glycine Proteins 0.000 description 1
- 108010070783 alanyltyrosine Proteins 0.000 description 1
- 230000003698 anagen phase Effects 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 108010091092 arginyl-glycyl-proline Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000005757 colony formation Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- NKLPQNGYXWVELD-UHFFFAOYSA-M coomassie brilliant blue Chemical compound [Na+].C1=CC(OCC)=CC=C1NC1=CC=C(C(=C2C=CC(C=C2)=[N+](CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=2C=CC(=CC=2)N(CC)CC=2C=C(C=CC=2)S([O-])(=O)=O)C=C1 NKLPQNGYXWVELD-UHFFFAOYSA-M 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011033 desalting Methods 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 210000001198 duodenum Anatomy 0.000 description 1
- 238000013399 early diagnosis Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- 210000002919 epithelial cell Anatomy 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 108010063718 gamma-glutamylaspartic acid Proteins 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 1
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 1
- 108010079413 glycyl-prolyl-glutamic acid Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 210000004013 groin Anatomy 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 238000002991 immunohistochemical analysis Methods 0.000 description 1
- 230000002055 immunohistochemical effect Effects 0.000 description 1
- 238000003364 immunohistochemistry Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 239000000411 inducer Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 239000002523 lectin Substances 0.000 description 1
- 108010057952 lysyl-phenylalanyl-lysine Proteins 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- FRZJZRVZZNTMAW-UHFFFAOYSA-N n,n-diethyl-3-(hydroxymethyl)benzamide Chemical compound CCN(CC)C(=O)C1=CC=CC(CO)=C1 FRZJZRVZZNTMAW-UHFFFAOYSA-N 0.000 description 1
- 229910052759 nickel Inorganic materials 0.000 description 1
- 210000000496 pancreas Anatomy 0.000 description 1
- 210000001819 pancreatic juice Anatomy 0.000 description 1
- 210000004923 pancreatic tissue Anatomy 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000003330 peritoneal dialysis fluid Substances 0.000 description 1
- 108010070409 phenylalanyl-glycyl-glycine Proteins 0.000 description 1
- 230000004962 physiological condition Effects 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000012257 pre-denaturation Methods 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000009465 prokaryotic expression Effects 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 108010079317 prolyl-tyrosine Proteins 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000001742 protein purification Methods 0.000 description 1
- 238000003127 radioimmunoassay Methods 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 108010033670 threonyl-aspartyl-tyrosine Proteins 0.000 description 1
- 238000000844 transformation Methods 0.000 description 1
- 230000001131 transforming effect Effects 0.000 description 1
- 108010044292 tryptophyltyrosine Proteins 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
Images
Classifications
-
- 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
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/577—Immunoassay; Biospecific binding assay; Materials therefor involving monoclonal antibodies binding reaction mechanisms characterised by the use of monoclonal antibodies; monoclonal antibodies per se are classified with their corresponding antigens
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- 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]
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2333/00—Assays involving biological materials from specific organisms or of a specific nature
- G01N2333/435—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans
- G01N2333/46—Assays involving biological materials from specific organisms or of a specific nature from animals; from humans from vertebrates
- G01N2333/47—Assays involving proteins of known structure or function as defined in the subgroups
- G01N2333/4701—Details
- G01N2333/474—Pancreatic thread protein; Reg protein
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Analytical Chemistry (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
本发明涉及再生蛋白REG1A单克隆抗体及其应用。该抗体包括重链和轻链,至少具有以下技术特征之一:i、重链包括重链CDR1,序列为VKQSH;ii、重链包括重链CDR2,序列为GGNVYNQKFKIKATLT;iii、重链包括重链CDR3,序列为PGTGYFDV;iv、轻链包括轻链CDR1,序列为RASQDISNYLN;v、轻链包括轻链CDR2,序列为YTSRLHS;vi、轻链包括轻链CDR3,序列为QQGHTLPRT。本发明的再生蛋白REG1A单克隆抗体能够有效检测再生蛋白REG1A水平,从而可以用以辅助诊断和治疗,以及预测疾病的发生和发展。
Description
技术领域
本发明涉及一种再生蛋白REG1A单克隆抗体及其应用,属于生物医药技术领域。
背景技术
再生基因(regenerating gene,Reg)是从再生胰岛中分离出来的一种基因,近年来,越来越多的研究表明再生基因家族蛋白的异常表达与炎症反应及肿瘤等多种疾病密切相关。其中再生蛋白(REG1)最早在大鼠再生胰岛中发现,后被证实与一种能够抑制胰腺结石形成的胰石蛋白(PSP)为同一种蛋白质,统称为PSP/reg。REG1A位于2p12染色体上,编码166个氨基酸,属于钙依赖性凝集素超家族。REG1A蛋白主要由胰腺腺泡细胞合成、分泌,是胰腺外分泌产物,生理状态下REG1A以可溶性形式主要分布于胰腺、消化道组织、胰液及血液中。REG1A具有促进增殖,诱导再生等作用,主要参与炎症、糖尿病、胰腺炎、肿瘤等疾病的发生。已有多项研究表明,REG1A的水平变化与这些疾病的进展密切相关,其在疾病的早期诊断及治疗中的作用越来越受到人们的重视。
经检索发现,申请号CN202010856716.7、申请公布号CN111909266A的发明专利申请公开了及一种再生基因蛋白REG1A单克隆抗体的制备方法,包括以下步骤:⑴制备免疫接种的小鼠;⑵骨髓瘤细胞培养3-4天,得到培养后的小鼠骨髓瘤细胞;⑶将REG1A蛋白溶解向免疫接种的小鼠腹腔接种免疫,并且获得脾细胞,经分散脾细胞得到脾细胞沉淀物;⑷培养后的小鼠骨髓瘤细胞离心得到骨髓瘤细胞沉淀物;⑸用DMEM高糖培养液分别悬浮脾细胞沉淀物、骨髓瘤细胞沉淀物并混合,经离心得到沉淀物;⑹沉淀物离心、悬浮,即得融合细胞;⑺将融合细胞进行培养、测定,确定杂交瘤细胞;⑻单抗的筛选分离;⑼大量制备单克隆抗体:将杂交瘤细胞系按常规方法进行单克隆抗体生产即得。然而,该技术方案并未能提供明确的单克隆抗体序列。
发明内容
本发明的主要目的是:克服现有技术存在的问题,提供一种再生蛋白REG1A单克隆抗体,能有效检测血清和组织中的REG1A;同时还提供该抗体的应用。
本发明解决其技术问题的技术方案如下:
一种再生蛋白REG1A单克隆抗体,所述抗体包括重链和轻链,其特征是,所述抗体至少具有以下技术特征之一:
i、所述重链包括重链CDR1,其氨基酸序列为VKQSH;
ii、所述重链包括重链CDR2,其氨基酸序列为GGNVYNQKFKIKATLT;
iii、所述重链包括重链CDR3,其氨基酸序列为PGTGYFDV;
iv、所述轻链包括轻链CDR1,其氨基酸序列为RASQDISNYLN;
v、所述轻链包括轻链CDR2,其氨基酸序列为YTSRLHS;
vi、所述轻链包括轻链CDR3,其氨基酸序列为QQGHTLPRT。
优选地,所述抗体至少具有以下技术特征之一:
i、所述重链包括重链CDR1,其氨基酸序列为VKQSH;所述重链包括重链CDR2,其氨基酸序列为GGNVYNQKFKIKATLT;所述重链包括重链CDR3,其氨基酸序列为PGTGYFDV;
ii、所述轻链包括轻链CDR1,其氨基酸序列为RASQDISNYLN;所述轻链包括轻链CDR2,其氨基酸序列为YTSRLHS;所述轻链包括轻链CDR3,其氨基酸序列为QQGHTLPRT。
优选地,所述抗体具有以下技术特征:
所述重链包括重链CDR1,其氨基酸序列为VKQSH;所述重链包括重链CDR2,其氨基酸序列为GGNVYNQKFKIKATLT;所述重链包括重链CDR3,其氨基酸序列为PGTGYFDV;
所述轻链包括轻链CDR1,其氨基酸序列为RASQDISNYLN;所述轻链包括轻链CDR2,其氨基酸序列为YTSRLHS;所述轻链包括轻链CDR3,其氨基酸序列为QQGHTLPRT。
优选地,所述重链的氨基酸序列如SEQ ID NO:1所示;所述轻链的氨基酸序列如SEQ ID NO:2所示。
优选地,所述抗体的重链或轻链上具有标记,所述标记包括荧光标记、酶标记、以及放射性标记。
本发明还提供:
编码前文所述再生蛋白REG1A单克隆抗体的核酸。
优选地,所述核酸包括编码重链的核酸且序列如SEQ ID NO:3所示,以及编码轻链的核酸且序列如SEQ ID NO:4所示。
本发明还提供:
前文所述再生蛋白REG1A单克隆抗体用于制备试剂盒的用途。
其中,所述试剂盒为针对再生蛋白REG1A的检测试剂盒或诊断试剂盒。
前文所述核酸用于制备再生蛋白REG1A单克隆抗体的用途。
本发明的再生蛋白REG1A单克隆抗体能够有效检测再生蛋白REG1A水平,从而可以用以辅助诊断和治疗,以及预测疾病的发生和发展。
附图说明
图1为本发明实施例1的SDS-PAGE分析结果图,其中1为纯化前样品,2为纯化后样品。
图2为本发明实施例1的Western blot分析结果图,其中1为REG1A重组蛋白,2为空载体质粒对照。
图3为本发明实施例2的单克隆抗体亚型鉴定分析结果图。
图4为本发明实施例2的Western blot分析结果图。
图5为本发明实施例2的细胞株单抗的效价检测结果图。
图6为本发明实施例2的再生蛋白REG1A单克隆抗体重链的具体可变区信息图。
图7为本发明实施例2的再生蛋白REG1A单克隆抗体轻链的具体可变区信息图。
图8为本发明实施例3的Western blot分析结果图。
图9为本发明实施例4的十二指肠免疫组化分析图。
具体实施方式
下面结合实施例对本发明作进一步详细描述。但是本发明不限于所给出的例子。
实施例1、原核表达带有组氨酸标签的重组再生蛋白REG1A。
1、获得目的基因人REG1A
通过本领域内常用方法,从人胰腺组织或人胰腺来源细胞系提取mRNA,逆转录得到cDNA,以编码REG1A蛋白的序列为基础设计特异性引物,在REG1A的N端添加6个组氨酸标签,并进行PCR扩增及扩增产物的琼脂糖凝胶电泳鉴定。
用于PCR扩增的引物为:
上游引物:5’-cagtggtggtggtggtggtgatggctcagaccagctcatacttc-3’
下游引物:5’-taccgacgacgacgacaaggctagtttttgaacttgcagacaaagg-3’
按以下程序进行PCR扩增:
(1)95℃预变性3min。(2)95℃变性15s。(3)58℃退火15s。(4)72℃延伸40s。(5)重复步骤(2)-(4)40次。(6)72℃延伸5min。
2、构建表达载体
(1)连接:以一种带有组氨酸标签的质粒为载体,如pET32a,经过PCR扩增得到载体cDNA。通过同源重组的方法连接线性化载体与目的基因片段,得到重组表达质粒pET32a-REG1A。
(2)转化:在冰上解冻克隆用的感受态细胞大肠杆菌DH5α。取适量重组质粒加入到感受态细胞中,冰上静置30min,后42℃水浴热激45s,立即置于冰上冷却2-3min;LB培养基(不含抗生素),37℃摇菌1小时(转速200-250rpm)。将含有氨苄抗性的LB平板在37℃培养箱中预热;菌液5000rpm离心5min,弃掉部分上清,重悬菌体,用无菌涂布棒在LB平板上轻涂均匀;在37℃培养箱中倒置培养12-16小时,长出的菌斑既为阳性克隆。
(3)筛选:挑选若干单克隆菌落,37℃摇菌3-4小时,后进行菌落PCR及基因测序鉴定。
3、表达菌株
将鉴定成功的菌液扩大培养,提取重组质粒pET32a-REG1A,并将其转化至表达蛋白的宿主菌体大肠杆菌BL21中,得到重组菌株BL21-REG1A。
4、重组蛋白的表达
将重组菌株接种于LB培养基中(含100mg/ml氨苄霉素),37℃,200rpm摇床过夜。次日按合适比例接种于新的LB培养基中,37℃,200rpm摇床培养至OD600为0.6-0.8,加入诱导剂IPTG继续培养。收集菌液,4℃,5000rpm离心20min,弃上清。经超声裂解菌体后分别取上清与沉淀进行12%SDS-PAGE电泳,考马斯亮蓝染色鉴定,确定重组蛋白HIS-REG1主要以可溶性蛋白形式存在于裂解液上清中。分别用不同温度、不同诱导剂浓度诱导菌体,确定融合蛋白表达的最佳条件,并进行蛋白的大量表达。
5、镍柱亲和层析法纯化蛋白
收集大量表达的菌体裂解液上清,过Ni琼脂糖凝胶亲和层析柱,收集洗脱峰流出液;使用超滤管(截留量10KDa)于4℃、4000×g离心20-30min进行浓缩、脱盐,得到纯化融合蛋白浓缩液。
将纯化前后的样品进行12%SDS-PAGE电泳,考马斯亮蓝染色鉴定,结果如图1所示,可见得到纯化的大小为19kD的REG1A重组蛋白。
6、REG1A重组蛋白的鉴定——鼠抗HIS标签单克隆抗体作为一抗的Western blot分析
将纯化后的REG1A重组蛋白进行SDS-PAGE后,将分离的蛋白从凝胶转至PVDF膜上,用5%脱脂奶粉室温封闭1h,后加入1:500稀释的鼠抗HIS标签单克隆抗体,4℃过夜。第二天TBST洗膜3次,加入辣根过氧化物酶(HRP)标记的羊抗鼠IgG抗体,37℃孵育1h,后PBST洗膜3次,进行ECL显色。显色结果(图2)显示在15-25kD范围内有大小约19kD的阳性单一条带,而转入空载体质粒的菌株无条带,表明REG1A重组蛋白成功表达。
实施例2、制备REG1A单克隆抗体。
1、制备融合蛋白REG1A的单克隆抗体
动物免疫:首次免疫取上述制备的带有组氨酸标签的REG1A重组蛋白,PBS稀释至合适比例,与完全福氏佐剂,以1:1的比例充分混合乳化。经颈背部、腹股沟皮下多点注射的方式,100mg/只,免疫小鼠3只,小鼠取6-8周的BALB/C鼠。两周后进行第二次免疫,取稀释的REG1A重组蛋白与不完全佐剂,以1:1的比例充分混合乳化,注射方式同上。两周后进行第三次免疫,方法同第二次免疫。三次免疫结束后取血清,其中即含有REG1A重组蛋白的多克隆抗体,经间接ELISA法或放免法可检测其效价。
2、细胞融合
①小鼠骨髓瘤细胞准备:SP2/0小鼠骨髓瘤细胞复苏,在含有8-氮杂鸟苷酸(20ug/mL)的无血清1640培养基中至少培养三代,以保持其HGPRT型。
②小鼠饲养细胞准备:细胞融合前1-2天,取6-8周龄BALC/B小鼠,腹腔注射不含血清的1640培养基,收集腹腔灌洗液,1000rpm离心5min,弃上清。加入HAT培养基重悬,将细胞稀释至1000个/ml,以100ul/孔将饲养细胞铺至96孔板中。
③小鼠脾细胞准备:细胞融合前3-4天对小鼠进行加强免疫,PBS稀释后的REG1A重组蛋白,腹腔注射,100mg/只。免疫当天取小鼠脾脏,制备脾细胞悬液,计数备用。
取对数生长期的小鼠骨髓瘤细胞SP2/0与脾细胞悬液,按4:1的比例将其与脾细胞混合,1000rpm离心5min,弃上清。轻轻弹匀试管底部的细胞沉淀。将试管置于37℃蒸馏水中,沿管壁缓慢加入1ml聚乙二醇融合剂,边加边缓慢转动混匀,1min内加完。后在3min内由慢到快加入3ml 37℃预热的1640培养基(未加血清)。重复上述步骤5min内加入20ml 37℃预热的1640培养基(未加血清),37℃水浴中静置10min。后1500rpm离心5min,弃上清,HAT培养基重悬,以100ul/孔加入到含有饲养细胞的96孔板中。培养箱中培养7-8天后观察细胞生长情况,之后2-3天进行一次半换液。
3、阳性杂交瘤细胞的筛选
当观察到有明显集落生成,且集落未融合前进行筛选。用REG1A重组蛋白为抗原,以0.1ug/ml浓度包被,取杂交瘤上清液100ul/孔进行间接ELISA检测,再在原培养板中不加HAT培养基,100ul/孔。
4、阳性杂交瘤的亚克隆
取阳性杂交瘤中OD值较高的孔,现转移至24孔板中扩大培养。用有限稀释的方法进行3-4次亚克隆,选取持续阳性、OD值高、形态好的单细胞亚克隆作为定植株,进行扩大培养和冻存保种。
5、制备单克隆抗体腹水
用体内诱生法制备单克隆抗体腹水,选取8周左右的BALB/C小鼠,向其腹腔内注射0.5ml石蜡油。将扩大培养的杂交瘤,吸去上清,将细胞重悬计数,以2×106个/mL,取0.5mL注射入小鼠腹腔。7天后收集腹水,即为REG1A单克隆抗体。
结果显示,A2、A4、A5、A6杂交瘤细胞株均可产生REG1A单克隆抗体。如图3所示,经单克隆抗体亚型鉴定分析,各REG1A单克隆抗体的重链均为IgG1型,且轻链为均为λ型。
6、以原核表达的无组氨酸标签的REG1A蛋白作为抗原,与A2、A4、A5、A6杂交瘤细胞株的REG1A单克隆抗体分别进行Western blot分析,结果如图4所示,所有细胞株的单克隆抗体均能与REG1A蛋白特异性结合,其中A6杂交瘤细胞株产生的抗体特异性相对较强,由此成功获得高特异性的REG1A单克隆抗体。
7、将A6细胞株杂交瘤扩大培养后腹腔注射入小鼠体内,收集小鼠腹水即得到REG1A单克隆抗体,用间接ELISA法进行效价检测,如图5所示,所得抗体的有效稀释倍速在128000以上。
8、将A6细胞株杂交瘤的REG1A单克隆抗体进行测序,结果如下:
(1)重链
氨基酸序列(SEQ ID NO:1):
SGPELVKPGASVKISCKASGYTFTDYNTHWVKQSHGKSLEWIGHIFPYNGGNVYNQKFKIKATLTVDISSSTAYMELRSLTSEDSAVYYCARRGPGTGYFDVWGAGTTVTVSS。
具体可变区信息如图6所示。
核酸序列(SEQ ID NO:3):
tcaggacctgagctggtgaaacctggggcctcagtgaagatctcctgcaaggcttctggctacacattcactgactataacacacactgggtgaagcagagccatggcaagagccttgagtggattggacatatttttccttacaatggtggtaatgtctacaaccagaagttcaagatcaaggccacattgactgtagacatttcctccagcacagcctacatggaactccgcagcctgacatctgaagactctgcagtctattactgtgcaagaaggggacctgggacggggtacttcgatgtctggggcgcagggaccacggtcaccgtctcctcag。
(2)轻链
氨基酸序列(SEQ ID NO:2):
DIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTSRLHSGVPSRFSGSGSGTDYSLTISNLDQEDIATYFCQQGHTLPRTFGGGTKLEIK。
具体可变区信息如图7所示。
核酸序列(SEQ ID NO:4):
gatatccagatgacacagactacatcctccctgtctgcctctctgggagacagagtcaccatcagttgcagggcaagtcaggacattagcaattatttaaactggtatcagcagaaaccagatggaactgttaaactcctgatctactacacatcaagattacactcaggagtcccatcaaggttcagtggcagtgggtctggaacagattattctctcaccattagcaacctggaccaagaagatattgccacttacttttgccaacagggtcatacacttcctcggacgttcggtggaggcaccaagctggaaatcaaac。
注:在具体实施时,可根据实际情况在抗体的重链或轻链上增设标记,如荧光标记、酶标记、放射性标记等。
实施例3、REG1A单克隆抗体的Western blot分析
现有相关研究已验证糖尿病患者血清中REG1A水平升高,以4例糖尿病患者血清为抗原,采用实施例2所制备的REG1A单克隆抗体作为一抗孵育显色后,结果如图8所示,REG1A单克隆抗体能够特异性识别血清中大小约19kD的REG1A蛋白。
实施例4、REG1A单克隆抗体的免疫组化分析
正常情况下REG1A在十二指肠组织中表达丰富。取正常的人十二指肠和肾组织的蜡片(注:上述蜡片为合法来源且符合伦理准则规定),经脱蜡、抗原修复后,以10%BSA室温封闭1h,PBS洗涤3次,加1:500稀释的实施例2所制备的REG1A单克隆抗体,4℃过夜。第二天PBS洗涤3次,加入辣根过氧化物酶(HRP)标记的兔抗鼠IgG抗体,室温孵育1h。PBS洗涤3次,后DAB显色,苏木素复染,封片剂封片。免疫组化结果显示,十二指肠上皮细胞包浆中可见棕色染色(图9),肾组织中未见,说明实施例2所制备的REG1A单克隆抗体可特异性识别正常组织中的REG1A蛋白。
除上述实施例外,本发明还可以有其他实施方式。凡采用等同替换或等效变换形成的技术方案,均落在本发明要求的保护范围。
序列表
<110> 东南大学附属中大医院
<120> 再生蛋白REG1A单克隆抗体及其应用
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 113
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
Ser Gly Pro Glu Leu Val Lys Pro Gly Ala Ser Val Lys Ile Ser Cys
1 5 10 15
Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr Asn Thr His Trp Val Lys
20 25 30
Gln Ser His Gly Lys Ser Leu Glu Trp Ile Gly His Ile Phe Pro Tyr
35 40 45
Asn Gly Gly Asn Val Tyr Asn Gln Lys Phe Lys Ile Lys Ala Thr Leu
50 55 60
Thr Val Asp Ile Ser Ser Ser Thr Ala Tyr Met Glu Leu Arg Ser Leu
65 70 75 80
Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys Ala Arg Arg Gly Pro Gly
85 90 95
Thr Gly Tyr Phe Asp Val Trp Gly Ala Gly Thr Thr Val Thr Val Ser
100 105 110
Ser
<210> 2
<211> 107
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly
1 5 10 15
Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr
20 25 30
Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile
35 40 45
Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly
50 55 60
Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Asp Gln
65 70 75 80
Glu Asp Ile Ala Thr Tyr Phe Cys Gln Gln Gly His Thr Leu Pro Arg
85 90 95
Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys
100 105
<210> 3
<211> 340
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
tcaggacctg agctggtgaa acctggggcc tcagtgaaga tctcctgcaa ggcttctggc 60
tacacattca ctgactataa cacacactgg gtgaagcaga gccatggcaa gagccttgag 120
tggattggac atatttttcc ttacaatggt ggtaatgtct acaaccagaa gttcaagatc 180
aaggccacat tgactgtaga catttcctcc agcacagcct acatggaact ccgcagcctg 240
acatctgaag actctgcagt ctattactgt gcaagaaggg gacctgggac ggggtacttc 300
gatgtctggg gcgcagggac cacggtcacc gtctcctcag 340
<210> 4
<211> 322
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 4
gatatccaga tgacacagac tacatcctcc ctgtctgcct ctctgggaga cagagtcacc 60
atcagttgca gggcaagtca ggacattagc aattatttaa actggtatca gcagaaacca 120
gatggaactg ttaaactcct gatctactac acatcaagat tacactcagg agtcccatca 180
aggttcagtg gcagtgggtc tggaacagat tattctctca ccattagcaa cctggaccaa 240
gaagatattg ccacttactt ttgccaacag ggtcatacac ttcctcggac gttcggtgga 300
ggcaccaagc tggaaatcaa ac 322
Claims (10)
1.一种再生蛋白REG1A单克隆抗体,所述抗体包括重链和轻链,其特征是,所述抗体至少具有以下技术特征之一:
i、所述重链包括重链CDR1,其氨基酸序列为VKQSH;
ii、所述重链包括重链CDR2,其氨基酸序列为GGNVYNQKFKIKATLT;
iii、所述重链包括重链CDR3,其氨基酸序列为PGTGYFDV;
iv、所述轻链包括轻链CDR1,其氨基酸序列为RASQDISNYLN;
v、所述轻链包括轻链CDR2,其氨基酸序列为YTSRLHS;
vi、所述轻链包括轻链CDR3,其氨基酸序列为QQGHTLPRT。
2.根据权利要求1所述的再生蛋白REG1A单克隆抗体,其特征是,所述抗体至少具有以下技术特征之一:
i、所述重链包括重链CDR1,其氨基酸序列为VKQSH;所述重链包括重链CDR2,其氨基酸序列为GGNVYNQKFKIKATLT;所述重链包括重链CDR3,其氨基酸序列为PGTGYFDV;
ii、所述轻链包括轻链CDR1,其氨基酸序列为RASQDISNYLN;所述轻链包括轻链CDR2,其氨基酸序列为YTSRLHS;所述轻链包括轻链CDR3,其氨基酸序列为QQGHTLPRT。
3.根据权利要求1所述的再生蛋白REG1A单克隆抗体,其特征是,所述抗体具有以下技术特征:
所述重链包括重链CDR1,其氨基酸序列为VKQSH;所述重链包括重链CDR2,其氨基酸序列为GGNVYNQKFKIKATLT;所述重链包括重链CDR3,其氨基酸序列为PGTGYFDV;
所述轻链包括轻链CDR1,其氨基酸序列为RASQDISNYLN;所述轻链包括轻链CDR2,其氨基酸序列为YTSRLHS;所述轻链包括轻链CDR3,其氨基酸序列为QQGHTLPRT。
4.根据权利要求1所述的再生蛋白REG1A单克隆抗体,其特征是,所述重链的氨基酸序列如SEQ ID NO:1所示;所述轻链的氨基酸序列如SEQ ID NO:2所示。
5.根据权利要求1至4任一项所述的再生蛋白REG1A单克隆抗体,其特征是,所述抗体的重链或轻链上具有标记,所述标记包括荧光标记、酶标记、以及放射性标记。
6.编码权利要求1至5任一项所述再生蛋白REG1A单克隆抗体的核酸。
7.根据权利要求6所述的核酸,其特征是,所述核酸包括编码重链的核酸且序列如SEQID NO:3所示,以及编码轻链的核酸且序列如SEQ ID NO:4所示。
8.权利要求1至5任一项所述再生蛋白REG1A单克隆抗体用于制备试剂盒的用途。
9.根据权利要求8所述的用途,其特征是,所述试剂盒为针对再生蛋白REG1A的检测试剂盒或诊断试剂盒。
10.权利要求6或7所述核酸用于制备再生蛋白REG1A单克隆抗体的用途。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210122652.7A CN114395040B (zh) | 2022-02-09 | 2022-02-09 | 再生蛋白reg1a单克隆抗体及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202210122652.7A CN114395040B (zh) | 2022-02-09 | 2022-02-09 | 再生蛋白reg1a单克隆抗体及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN114395040A true CN114395040A (zh) | 2022-04-26 |
CN114395040B CN114395040B (zh) | 2023-12-15 |
Family
ID=81233705
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210122652.7A Active CN114395040B (zh) | 2022-02-09 | 2022-02-09 | 再生蛋白reg1a单克隆抗体及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN114395040B (zh) |
Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0369797A2 (en) * | 1988-11-16 | 1990-05-23 | SHIONOGI SEIYAKU KABUSHIKI KAISHA trading under the name of SHIONOGI & CO. LTD. | A monoclonal antibody recognizing a reg protein |
WO1998016640A1 (en) * | 1996-10-11 | 1998-04-23 | Incyte Pharmaceuticals, Inc. | Human reg i-gamma protein |
CN101270161A (zh) * | 2008-03-07 | 2008-09-24 | 浙江大学 | 抗人Reg4的单克隆抗体及其制备与应用以及杂交瘤细胞株 |
CN111909266A (zh) * | 2020-08-24 | 2020-11-10 | 甘肃智选生物科技有限公司 | 一种再生基因蛋白REG1α单克隆抗体的制备方法 |
CN113248609A (zh) * | 2021-07-14 | 2021-08-13 | 深圳市盛波尔生命科学技术有限责任公司 | 针对再生胰岛衍生蛋白1α的抗体组合以及包含其的检测试剂盒 |
CN113423735A (zh) * | 2018-12-07 | 2021-09-21 | 再鼎医药(上海)有限公司 | 抗-紧密连接蛋白抗体及其用途 |
-
2022
- 2022-02-09 CN CN202210122652.7A patent/CN114395040B/zh active Active
Patent Citations (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP0369797A2 (en) * | 1988-11-16 | 1990-05-23 | SHIONOGI SEIYAKU KABUSHIKI KAISHA trading under the name of SHIONOGI & CO. LTD. | A monoclonal antibody recognizing a reg protein |
WO1998016640A1 (en) * | 1996-10-11 | 1998-04-23 | Incyte Pharmaceuticals, Inc. | Human reg i-gamma protein |
CN101270161A (zh) * | 2008-03-07 | 2008-09-24 | 浙江大学 | 抗人Reg4的单克隆抗体及其制备与应用以及杂交瘤细胞株 |
CN113423735A (zh) * | 2018-12-07 | 2021-09-21 | 再鼎医药(上海)有限公司 | 抗-紧密连接蛋白抗体及其用途 |
CN111909266A (zh) * | 2020-08-24 | 2020-11-10 | 甘肃智选生物科技有限公司 | 一种再生基因蛋白REG1α单克隆抗体的制备方法 |
CN113248609A (zh) * | 2021-07-14 | 2021-08-13 | 深圳市盛波尔生命科学技术有限责任公司 | 针对再生胰岛衍生蛋白1α的抗体组合以及包含其的检测试剂盒 |
Non-Patent Citations (1)
Title |
---|
雷有玲;李国泰;鲍玉林;刘秀清;张文;: ""猪伪狂犬病毒gE蛋白单克隆抗体制备与鉴定"", 《畜牧与兽医》, no. 01, pages 115 - 118 * |
Also Published As
Publication number | Publication date |
---|---|
CN114395040B (zh) | 2023-12-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP7062626B2 (ja) | 組換えDsbCを発現する細菌宿主系統 | |
JP5289285B2 (ja) | Vegf−特異的なヒト抗体 | |
CN113150136B (zh) | 新型冠状病毒n蛋白单克隆抗体的制备 | |
WO2017121307A1 (zh) | 一种C5aR抗体及其制备方法和应用 | |
CN111057148B (zh) | 针对牛血清白蛋白bsa的单域抗体及其衍生蛋白 | |
CN111018985B (zh) | 针对牛血清白蛋白bsa的单域抗体的应用 | |
CN110922489B (zh) | 一种抗C3d的靶向单链抗体和CD59的融合蛋白及应用 | |
CN112661849B (zh) | 一种艰难梭菌重组蛋白单克隆抗体的制备方法和应用 | |
CN110922480A (zh) | 一种抗C3d的靶向单链抗体和DAF的融合蛋白及应用 | |
CN116162160A (zh) | 一种抗il-6的单域抗体及其用途 | |
CN113150138B (zh) | 一种kpc-2单克隆抗体及其制备方法和应用 | |
KR102453605B1 (ko) | 신규 펩타이드 태그, 이에 결합하는 항체 및 이들의 용도 | |
CN108884153B (zh) | 与aimp2-dx2蛋白质特异性结合的抗体 | |
CN109021103B (zh) | 抗人血管内皮生长因子的抗体及其制备方法和应用 | |
WO2015113494A1 (zh) | 双功能融合蛋白及其制备方法和用途 | |
CN110511277B (zh) | 一种抗hsp90单克隆抗体及其应用 | |
CN114395040B (zh) | 再生蛋白reg1a单克隆抗体及其应用 | |
CN107686519B (zh) | 抗小鼠mxra7单克隆抗体的制备方法及其应用 | |
CN113150139B (zh) | 一种PBP2a单克隆抗体及其制备方法和应用 | |
CN110885375B (zh) | 特异性针对mmp-9蛋白锌离子结合结构域的单域抗体及产品与应用 | |
CN111763255B (zh) | 一种经基因修饰的vegfa蛋白及其单克隆抗体与应用 | |
CN110894236B (zh) | 一种抗曲霉菌半乳甘露聚糖的单克隆抗体及其应用 | |
CN110894235B (zh) | 一种抗隐球菌夹膜多糖的兔源单克隆抗体及其应用 | |
CN114699507A (zh) | 一种细胞外Ezrin蛋白的应用 | |
CN113912729B (zh) | 针对sST2的单域抗体及其衍生蛋白和应用 |
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 | ||
GR01 | Patent grant | ||
GR01 | Patent grant |