CN114609394A - Anti-mullerian tube hormone detection kit, monoclonal antibody and hybridoma cell strain - Google Patents
Anti-mullerian tube hormone detection kit, monoclonal antibody and hybridoma cell strain Download PDFInfo
- Publication number
- CN114609394A CN114609394A CN202210324182.2A CN202210324182A CN114609394A CN 114609394 A CN114609394 A CN 114609394A CN 202210324182 A CN202210324182 A CN 202210324182A CN 114609394 A CN114609394 A CN 114609394A
- Authority
- CN
- China
- Prior art keywords
- monoclonal antibody
- gly
- hybridoma cell
- antibody
- leu
- 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
- 210000004408 hybridoma Anatomy 0.000 title claims abstract description 38
- 238000001514 detection method Methods 0.000 title claims abstract description 34
- 239000005556 hormone Substances 0.000 title description 2
- 229940088597 hormone Drugs 0.000 title description 2
- 108010005853 Anti-Mullerian Hormone Proteins 0.000 claims abstract description 63
- 102100030173 Muellerian-inhibiting factor Human genes 0.000 claims abstract description 63
- 239000000868 anti-mullerian hormone Substances 0.000 claims abstract description 62
- 238000004321 preservation Methods 0.000 claims abstract description 30
- 108090000765 processed proteins & peptides Proteins 0.000 claims description 40
- 229920001184 polypeptide Polymers 0.000 claims description 39
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 39
- 238000003018 immunoassay Methods 0.000 claims description 13
- 238000012216 screening Methods 0.000 claims description 7
- DKJWUIYLMLUBDX-XPUUQOCRSA-N Gly-Val-Cys Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CS)C(=O)O DKJWUIYLMLUBDX-XPUUQOCRSA-N 0.000 claims description 5
- 239000011324 bead Substances 0.000 claims description 5
- 108010078144 glutaminyl-glycine Proteins 0.000 claims description 5
- MFAMTAVAFBPXDC-LPEHRKFASA-N Arg-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)C(=O)O MFAMTAVAFBPXDC-LPEHRKFASA-N 0.000 claims description 4
- SCQBNMKLZVCXNX-ZFWWWQNUSA-N Trp-Arg-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)NCC(=O)O)N SCQBNMKLZVCXNX-ZFWWWQNUSA-N 0.000 claims description 4
- 108010091092 arginyl-glycyl-proline Proteins 0.000 claims description 4
- 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 claims description 3
- ULBHWNVWSCJLCO-NHCYSSNCSA-N Arg-Val-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CCCN=C(N)N ULBHWNVWSCJLCO-NHCYSSNCSA-N 0.000 claims description 3
- TWVTVZUGEDBAJF-ACZMJKKPSA-N Asn-Cys-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC(=O)N)N TWVTVZUGEDBAJF-ACZMJKKPSA-N 0.000 claims description 3
- WUAYFMZULZDSLB-ACZMJKKPSA-N Gln-Ala-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O WUAYFMZULZDSLB-ACZMJKKPSA-N 0.000 claims description 3
- UTOQQOMEJDPDMX-ACZMJKKPSA-N Gln-Ser-Asp Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O UTOQQOMEJDPDMX-ACZMJKKPSA-N 0.000 claims description 3
- VHPVBPCCWVDGJL-IRIUXVKKSA-N Glu-Thr-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O VHPVBPCCWVDGJL-IRIUXVKKSA-N 0.000 claims description 3
- JXYMPBCYRKWJEE-BQBZGAKWSA-N Gly-Arg-Ala Chemical compound [H]NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O JXYMPBCYRKWJEE-BQBZGAKWSA-N 0.000 claims description 3
- PASHZZBXZYEXFE-LSDHHAIUSA-N Gly-Trp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CNC3=CC=CC=C32)NC(=O)CN)C(=O)O PASHZZBXZYEXFE-LSDHHAIUSA-N 0.000 claims description 3
- DPWGZWUMUUJQDT-IUCAKERBSA-N Leu-Gln-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O DPWGZWUMUUJQDT-IUCAKERBSA-N 0.000 claims description 3
- 108010044940 alanylglutamine Proteins 0.000 claims description 3
- 108010050025 alpha-glutamyltryptophan Proteins 0.000 claims description 3
- 108010016616 cysteinylglycine Proteins 0.000 claims description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 21
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 18
- 238000012360 testing method Methods 0.000 description 18
- 239000000427 antigen Substances 0.000 description 15
- 102000036639 antigens Human genes 0.000 description 15
- 108091007433 antigens Proteins 0.000 description 15
- 239000000523 sample Substances 0.000 description 15
- 238000000034 method Methods 0.000 description 13
- 238000002156 mixing Methods 0.000 description 13
- 238000002360 preparation method Methods 0.000 description 13
- 210000002966 serum Anatomy 0.000 description 12
- 238000005406 washing Methods 0.000 description 12
- 229960002685 biotin Drugs 0.000 description 9
- 235000020958 biotin Nutrition 0.000 description 9
- 239000011616 biotin Substances 0.000 description 9
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 8
- 239000008055 phosphate buffer solution Substances 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 210000004027 cell Anatomy 0.000 description 7
- 238000007865 diluting Methods 0.000 description 7
- 230000003053 immunization Effects 0.000 description 7
- 238000002965 ELISA Methods 0.000 description 6
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 6
- 210000004899 c-terminal region Anatomy 0.000 description 6
- 230000008878 coupling Effects 0.000 description 6
- 238000010168 coupling process Methods 0.000 description 6
- 238000005859 coupling reaction Methods 0.000 description 6
- 238000002649 immunization Methods 0.000 description 6
- 239000007790 solid phase Substances 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 238000002372 labelling Methods 0.000 description 5
- 230000002611 ovarian Effects 0.000 description 5
- 230000002194 synthesizing effect Effects 0.000 description 5
- PXHVJJICTQNCMI-UHFFFAOYSA-N Nickel Chemical compound [Ni] PXHVJJICTQNCMI-UHFFFAOYSA-N 0.000 description 4
- 238000004140 cleaning Methods 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 238000009629 microbiological culture Methods 0.000 description 4
- 239000013642 negative control Substances 0.000 description 4
- 108090000623 proteins and genes Proteins 0.000 description 4
- 108010032595 Antibody Binding Sites Proteins 0.000 description 3
- YBZMTKUDWXZLIX-UWVGGRQHSA-N Arg-Leu-Gly Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O YBZMTKUDWXZLIX-UWVGGRQHSA-N 0.000 description 3
- 238000009007 Diagnostic Kit Methods 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 241000699670 Mus sp. Species 0.000 description 3
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 3
- POQFNPILEQEODH-FXQIFTODSA-N Pro-Ser-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O POQFNPILEQEODH-FXQIFTODSA-N 0.000 description 3
- 108010043240 arginyl-leucyl-glycine Proteins 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000011248 coating agent Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 238000000502 dialysis Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 108010020755 prolyl-glycyl-glycine Proteins 0.000 description 3
- 238000013112 stability test Methods 0.000 description 3
- VGPWRRFOPXVGOH-BYPYZUCNSA-N Ala-Gly-Gly Chemical compound C[C@H](N)C(=O)NCC(=O)NCC(O)=O VGPWRRFOPXVGOH-BYPYZUCNSA-N 0.000 description 2
- KXOPYFNQLVUOAQ-FXQIFTODSA-N Arg-Ser-Ala Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O KXOPYFNQLVUOAQ-FXQIFTODSA-N 0.000 description 2
- FMNBYVSGRCXWEK-FOHZUACHSA-N Asn-Thr-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O FMNBYVSGRCXWEK-FOHZUACHSA-N 0.000 description 2
- QQAYIVHVRFJICE-AEJSXWLSSA-N Cys-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CS)N QQAYIVHVRFJICE-AEJSXWLSSA-N 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 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 2
- QPTNELDXWKRIFX-YFKPBYRVSA-N Gly-Gly-Gln Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCC(N)=O QPTNELDXWKRIFX-YFKPBYRVSA-N 0.000 description 2
- 108090000288 Glycoproteins Proteins 0.000 description 2
- 102000003886 Glycoproteins Human genes 0.000 description 2
- 101000945318 Homo sapiens Calponin-1 Proteins 0.000 description 2
- 101000652736 Homo sapiens Transgelin Proteins 0.000 description 2
- WNGVUZWBXZKQES-YUMQZZPRSA-N Leu-Ala-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O WNGVUZWBXZKQES-YUMQZZPRSA-N 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 206010036049 Polycystic ovaries Diseases 0.000 description 2
- 208000002500 Primary Ovarian Insufficiency Diseases 0.000 description 2
- CGBYDGAJHSOGFQ-LPEHRKFASA-N Pro-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2 CGBYDGAJHSOGFQ-LPEHRKFASA-N 0.000 description 2
- SBVPYBFMIGDIDX-SRVKXCTJSA-N Pro-Pro-Pro Chemical compound OC(=O)[C@@H]1CCCN1C(=O)[C@H]1N(C(=O)[C@H]2NCCC2)CCC1 SBVPYBFMIGDIDX-SRVKXCTJSA-N 0.000 description 2
- GZFAWAQTEYDKII-YUMQZZPRSA-N Ser-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CO GZFAWAQTEYDKII-YUMQZZPRSA-N 0.000 description 2
- QYSFWUIXDFJUDW-DCAQKATOSA-N Ser-Leu-Arg Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O QYSFWUIXDFJUDW-DCAQKATOSA-N 0.000 description 2
- YEDSOSIKVUMIJE-DCAQKATOSA-N Ser-Val-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O YEDSOSIKVUMIJE-DCAQKATOSA-N 0.000 description 2
- 102100031013 Transgelin Human genes 0.000 description 2
- MNSSBIHFEUUXNW-RCWTZXSCSA-N Val-Thr-Arg Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CCCN=C(N)N MNSSBIHFEUUXNW-RCWTZXSCSA-N 0.000 description 2
- DZBUGLKDJFMEHC-UHFFFAOYSA-N acridine Chemical compound C1=CC=CC2=CC3=CC=CC=C3N=C21 DZBUGLKDJFMEHC-UHFFFAOYSA-N 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 238000001261 affinity purification Methods 0.000 description 2
- 108010005233 alanylglutamic acid Proteins 0.000 description 2
- 108010047495 alanylglycine Proteins 0.000 description 2
- KOSRFJWDECSPRO-UHFFFAOYSA-N alpha-L-glutamyl-L-glutamic acid Natural products OC(=O)CCC(N)C(=O)NC(CCC(O)=O)C(O)=O KOSRFJWDECSPRO-UHFFFAOYSA-N 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 108010093581 aspartyl-proline Proteins 0.000 description 2
- 239000012496 blank sample Substances 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 239000012888 bovine serum Substances 0.000 description 2
- 239000007853 buffer solution Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000010612 desalination reaction Methods 0.000 description 2
- 238000011033 desalting Methods 0.000 description 2
- 239000000539 dimer Substances 0.000 description 2
- 239000013604 expression vector Substances 0.000 description 2
- VPZXBVLAVMBEQI-UHFFFAOYSA-N glycyl-DL-alpha-alanine Natural products OC(=O)C(C)NC(=O)CN VPZXBVLAVMBEQI-UHFFFAOYSA-N 0.000 description 2
- XKUKSGPZAADMRA-UHFFFAOYSA-N glycyl-glycyl-glycine Natural products NCC(=O)NCC(=O)NCC(O)=O XKUKSGPZAADMRA-UHFFFAOYSA-N 0.000 description 2
- 230000036046 immunoreaction Effects 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000004020 luminiscence type Methods 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 229910052759 nickel Inorganic materials 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 201000010065 polycystic ovary syndrome Diseases 0.000 description 2
- 206010036601 premature menopause Diseases 0.000 description 2
- 208000017942 premature ovarian failure 1 Diseases 0.000 description 2
- 108010090894 prolylleucine Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 210000004988 splenocyte Anatomy 0.000 description 2
- JJAHTWIKCUJRDK-UHFFFAOYSA-N succinimidyl 4-(N-maleimidomethyl)cyclohexane-1-carboxylate Chemical compound C1CC(CN2C(C=CC2=O)=O)CCC1C(=O)ON1C(=O)CCC1=O JJAHTWIKCUJRDK-UHFFFAOYSA-N 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- DTLVBHCSSNJCMJ-UHFFFAOYSA-N (2,5-dioxopyrrolidin-1-yl) 3-[2-[2-[2-[2-[5-(2-oxo-1,3,3a,4,6,6a-hexahydrothieno[3,4-d]imidazol-4-yl)pentanoylamino]ethoxy]ethoxy]ethoxy]ethoxy]propanoate Chemical compound S1CC2NC(=O)NC2C1CCCCC(=O)NCCOCCOCCOCCOCCC(=O)ON1C(=O)CCC1=O DTLVBHCSSNJCMJ-UHFFFAOYSA-N 0.000 description 1
- XVZCXCTYGHPNEM-IHRRRGAJSA-N (2s)-1-[(2s)-2-[[(2s)-2-amino-4-methylpentanoyl]amino]-4-methylpentanoyl]pyrrolidine-2-carboxylic acid Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(O)=O XVZCXCTYGHPNEM-IHRRRGAJSA-N 0.000 description 1
- PKOHVHWNGUHYRE-ZFWWWQNUSA-N (2s)-1-[2-[[(2s)-2-amino-3-(1h-indol-3-yl)propanoyl]amino]acetyl]pyrrolidine-2-carboxylic acid Chemical compound O=C([C@H](CC=1C2=CC=CC=C2NC=1)N)NCC(=O)N1CCC[C@H]1C(O)=O PKOHVHWNGUHYRE-ZFWWWQNUSA-N 0.000 description 1
- BRPMXFSTKXXNHF-IUCAKERBSA-N (2s)-1-[2-[[(2s)-pyrrolidine-2-carbonyl]amino]acetyl]pyrrolidine-2-carboxylic acid Chemical compound OC(=O)[C@@H]1CCCN1C(=O)CNC(=O)[C@H]1NCCC1 BRPMXFSTKXXNHF-IUCAKERBSA-N 0.000 description 1
- 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
- WOJJIRYPFAZEPF-YFKPBYRVSA-N 2-[[(2s)-2-[[2-[(2-azaniumylacetyl)amino]acetyl]amino]propanoyl]amino]acetate Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)CNC(=O)CN WOJJIRYPFAZEPF-YFKPBYRVSA-N 0.000 description 1
- QMOQBVOBWVNSNO-UHFFFAOYSA-N 2-[[2-[[2-[(2-azaniumylacetyl)amino]acetyl]amino]acetyl]amino]acetate Chemical compound NCC(=O)NCC(=O)NCC(=O)NCC(O)=O QMOQBVOBWVNSNO-UHFFFAOYSA-N 0.000 description 1
- JBVSSSZFNTXJDX-YTLHQDLWSA-N Ala-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@H](C)N JBVSSSZFNTXJDX-YTLHQDLWSA-N 0.000 description 1
- SDMAQFGBPOJFOM-GUBZILKMSA-N Ala-Arg-Arg Chemical compound NC(=N)NCCC[C@H](NC(=O)[C@@H](N)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SDMAQFGBPOJFOM-GUBZILKMSA-N 0.000 description 1
- LBJYAILUMSUTAM-ZLUOBGJFSA-N Ala-Asn-Asn Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O LBJYAILUMSUTAM-ZLUOBGJFSA-N 0.000 description 1
- KIUYPHAMDKDICO-WHFBIAKZSA-N Ala-Asp-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O KIUYPHAMDKDICO-WHFBIAKZSA-N 0.000 description 1
- NWVVKQZOVSTDBQ-CIUDSAMLSA-N Ala-Glu-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NWVVKQZOVSTDBQ-CIUDSAMLSA-N 0.000 description 1
- ZVFVBBGVOILKPO-WHFBIAKZSA-N Ala-Gly-Ala Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O ZVFVBBGVOILKPO-WHFBIAKZSA-N 0.000 description 1
- PCIFXPRIFWKWLK-YUMQZZPRSA-N Ala-Gly-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@H](C)N PCIFXPRIFWKWLK-YUMQZZPRSA-N 0.000 description 1
- BLIMFWGRQKRCGT-YUMQZZPRSA-N Ala-Gly-Lys Chemical compound C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN BLIMFWGRQKRCGT-YUMQZZPRSA-N 0.000 description 1
- 108010076441 Ala-His-His Proteins 0.000 description 1
- SUMYEVXWCAYLLJ-GUBZILKMSA-N Ala-Leu-Gln Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O SUMYEVXWCAYLLJ-GUBZILKMSA-N 0.000 description 1
- MNZHHDPWDWQJCQ-YUMQZZPRSA-N Ala-Leu-Gly Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O MNZHHDPWDWQJCQ-YUMQZZPRSA-N 0.000 description 1
- OYJCVIGKMXUVKB-GARJFASQSA-N Ala-Leu-Pro Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N1CCC[C@@H]1C(=O)O)N OYJCVIGKMXUVKB-GARJFASQSA-N 0.000 description 1
- SOBIAADAMRHGKH-CIUDSAMLSA-N Ala-Leu-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(O)=O SOBIAADAMRHGKH-CIUDSAMLSA-N 0.000 description 1
- MEFILNJXAVSUTO-JXUBOQSCSA-N Ala-Leu-Thr Chemical compound C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MEFILNJXAVSUTO-JXUBOQSCSA-N 0.000 description 1
- ADSGHMXEAZJJNF-DCAQKATOSA-N Ala-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](C)N ADSGHMXEAZJJNF-DCAQKATOSA-N 0.000 description 1
- LSMDIAAALJJLRO-XQXXSGGOSA-N Ala-Thr-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(O)=O LSMDIAAALJJLRO-XQXXSGGOSA-N 0.000 description 1
- JJHBEVZAZXZREW-LFSVMHDDSA-N Ala-Thr-Phe Chemical compound C[C@@H](O)[C@H](NC(=O)[C@H](C)N)C(=O)N[C@@H](Cc1ccccc1)C(O)=O JJHBEVZAZXZREW-LFSVMHDDSA-N 0.000 description 1
- YXXPVUOMPSZURS-ZLIFDBKOSA-N Ala-Trp-Leu Chemical compound C1=CC=C2C(C[C@@H](C(=O)N[C@@H](CC(C)C)C(O)=O)NC(=O)[C@H](C)N)=CNC2=C1 YXXPVUOMPSZURS-ZLIFDBKOSA-N 0.000 description 1
- BGGAIXWIZCIFSG-XDTLVQLUSA-N Ala-Tyr-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O BGGAIXWIZCIFSG-XDTLVQLUSA-N 0.000 description 1
- GXCSUJQOECMKPV-CIUDSAMLSA-N Arg-Ala-Gln Chemical compound C[C@H](NC(=O)[C@@H](N)CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O GXCSUJQOECMKPV-CIUDSAMLSA-N 0.000 description 1
- VKKYFICVTYKFIO-CIUDSAMLSA-N Arg-Ala-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N VKKYFICVTYKFIO-CIUDSAMLSA-N 0.000 description 1
- OTCJMMRQBVDQRK-DCAQKATOSA-N Arg-Asp-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O OTCJMMRQBVDQRK-DCAQKATOSA-N 0.000 description 1
- 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 1
- AWMAZIIEFPFHCP-RCWTZXSCSA-N Arg-Pro-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O AWMAZIIEFPFHCP-RCWTZXSCSA-N 0.000 description 1
- ISJWBVIYRBAXEB-CIUDSAMLSA-N Arg-Ser-Glu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(O)=O ISJWBVIYRBAXEB-CIUDSAMLSA-N 0.000 description 1
- 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 1
- 206010003445 Ascites Diseases 0.000 description 1
- FTCGGKNCJZOPNB-WHFBIAKZSA-N Asn-Gly-Ser Chemical compound NC(=O)C[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O FTCGGKNCJZOPNB-WHFBIAKZSA-N 0.000 description 1
- WIDVAWAQBRAKTI-YUMQZZPRSA-N Asn-Leu-Gly Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O WIDVAWAQBRAKTI-YUMQZZPRSA-N 0.000 description 1
- PBFXCUOEGVJTMV-QXEWZRGKSA-N Asn-Met-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C(C)C)C(O)=O PBFXCUOEGVJTMV-QXEWZRGKSA-N 0.000 description 1
- QHAJMRDEWNAIBQ-FXQIFTODSA-N Asp-Arg-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O QHAJMRDEWNAIBQ-FXQIFTODSA-N 0.000 description 1
- SOYOSFXLXYZNRG-CIUDSAMLSA-N Asp-Arg-Gln Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O SOYOSFXLXYZNRG-CIUDSAMLSA-N 0.000 description 1
- DBWYWXNMZZYIRY-LPEHRKFASA-N Asp-Arg-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(=O)O)N)C(=O)O DBWYWXNMZZYIRY-LPEHRKFASA-N 0.000 description 1
- SCQIQCWLOMOEFP-DCAQKATOSA-N Asp-Leu-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O SCQIQCWLOMOEFP-DCAQKATOSA-N 0.000 description 1
- PAYPSKIBMDHZPI-CIUDSAMLSA-N Asp-Leu-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O PAYPSKIBMDHZPI-CIUDSAMLSA-N 0.000 description 1
- ZKAOJVJQGVUIIU-GUBZILKMSA-N Asp-Pro-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O ZKAOJVJQGVUIIU-GUBZILKMSA-N 0.000 description 1
- XUVTWGPERWIERB-IHRRRGAJSA-N Asp-Pro-Phe Chemical compound N[C@@H](CC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1ccccc1)C(O)=O XUVTWGPERWIERB-IHRRRGAJSA-N 0.000 description 1
- MJJIHRWNWSQTOI-VEVYYDQMSA-N Asp-Thr-Arg Chemical compound OC(=O)C[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O MJJIHRWNWSQTOI-VEVYYDQMSA-N 0.000 description 1
- ITGFVUYOLWBPQW-KKHAAJSZSA-N Asp-Thr-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O ITGFVUYOLWBPQW-KKHAAJSZSA-N 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- KEBJBKIASQVRJS-WDSKDSINSA-N Cys-Gln-Gly Chemical compound C(CC(=O)N)[C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CS)N KEBJBKIASQVRJS-WDSKDSINSA-N 0.000 description 1
- JDHMXPSXWMPYQZ-AAEUAGOBSA-N Cys-Gly-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CS)N JDHMXPSXWMPYQZ-AAEUAGOBSA-N 0.000 description 1
- SWJYSDXMTPMBHO-FXQIFTODSA-N Cys-Pro-Ser Chemical compound [H]N[C@@H](CS)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CO)C(O)=O SWJYSDXMTPMBHO-FXQIFTODSA-N 0.000 description 1
- BWGNESOTFCXPMA-UHFFFAOYSA-N Dihydrogen disulfide Chemical compound SS BWGNESOTFCXPMA-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- XUMFMAVDHQDATI-DCAQKATOSA-N Gln-Pro-Arg Chemical compound NC(=O)CC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O XUMFMAVDHQDATI-DCAQKATOSA-N 0.000 description 1
- LPIKVBWNNVFHCQ-GUBZILKMSA-N Gln-Ser-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O LPIKVBWNNVFHCQ-GUBZILKMSA-N 0.000 description 1
- VTTSANCGJWLPNC-ZPFDUUQYSA-N Glu-Arg-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O VTTSANCGJWLPNC-ZPFDUUQYSA-N 0.000 description 1
- IRXNJYPKBVERCW-DCAQKATOSA-N Glu-Leu-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IRXNJYPKBVERCW-DCAQKATOSA-N 0.000 description 1
- ATVYZJGOZLVXDK-IUCAKERBSA-N Glu-Leu-Gly Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)NCC(O)=O ATVYZJGOZLVXDK-IUCAKERBSA-N 0.000 description 1
- QJVZSVUYZFYLFQ-CIUDSAMLSA-N Glu-Pro-Ala Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O QJVZSVUYZFYLFQ-CIUDSAMLSA-N 0.000 description 1
- HMJULNMJWOZNFI-XHNCKOQMSA-N Glu-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)O)N)C(=O)O HMJULNMJWOZNFI-XHNCKOQMSA-N 0.000 description 1
- WGYHAAXZWPEBDQ-IFFSRLJSSA-N Glu-Val-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WGYHAAXZWPEBDQ-IFFSRLJSSA-N 0.000 description 1
- PYTZFYUXZZHOAD-WHFBIAKZSA-N Gly-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)CN PYTZFYUXZZHOAD-WHFBIAKZSA-N 0.000 description 1
- 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 1
- GGEJHJIXRBTJPD-BYPYZUCNSA-N Gly-Asn-Gly Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GGEJHJIXRBTJPD-BYPYZUCNSA-N 0.000 description 1
- LCNXZQROPKFGQK-WHFBIAKZSA-N Gly-Asp-Ser Chemical compound NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O LCNXZQROPKFGQK-WHFBIAKZSA-N 0.000 description 1
- YZACQYVWLCQWBT-BQBZGAKWSA-N Gly-Cys-Arg Chemical compound [H]NCC(=O)N[C@@H](CS)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O YZACQYVWLCQWBT-BQBZGAKWSA-N 0.000 description 1
- FIQQRCFQXGLOSZ-WDSKDSINSA-N Gly-Glu-Asp Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O FIQQRCFQXGLOSZ-WDSKDSINSA-N 0.000 description 1
- IDOGEHIWMJMAHT-BYPYZUCNSA-N Gly-Gly-Cys Chemical compound NCC(=O)NCC(=O)N[C@@H](CS)C(O)=O IDOGEHIWMJMAHT-BYPYZUCNSA-N 0.000 description 1
- NSTUFLGQJCOCDL-UWVGGRQHSA-N Gly-Leu-Arg Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N NSTUFLGQJCOCDL-UWVGGRQHSA-N 0.000 description 1
- UUYBFNKHOCJCHT-VHSXEESVSA-N Gly-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN UUYBFNKHOCJCHT-VHSXEESVSA-N 0.000 description 1
- 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 1
- NSVOVKWEKGEOQB-LURJTMIESA-N Gly-Pro-Gly Chemical compound NCC(=O)N1CCC[C@H]1C(=O)NCC(O)=O NSVOVKWEKGEOQB-LURJTMIESA-N 0.000 description 1
- ABPRMMYHROQBLY-NKWVEPMBSA-N Gly-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)CN)C(=O)O ABPRMMYHROQBLY-NKWVEPMBSA-N 0.000 description 1
- JBCLFWXMTIKCCB-UHFFFAOYSA-N H-Gly-Phe-OH Natural products NCC(=O)NC(C(O)=O)CC1=CC=CC=C1 JBCLFWXMTIKCCB-UHFFFAOYSA-N 0.000 description 1
- CHZRWFUGWRTUOD-IUCAKERBSA-N His-Gly-Gln Chemical compound C1=C(NC=N1)C[C@@H](C(=O)NCC(=O)N[C@@H](CCC(=O)N)C(=O)O)N CHZRWFUGWRTUOD-IUCAKERBSA-N 0.000 description 1
- FBOMZVOKCZMDIG-XQQFMLRXSA-N His-Val-Pro Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CN=CN2)N FBOMZVOKCZMDIG-XQQFMLRXSA-N 0.000 description 1
- UYNXBNHVWFNVIN-HJWJTTGWSA-N Ile-Phe-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)[C@@H](C)CC)CC1=CC=CC=C1 UYNXBNHVWFNVIN-HJWJTTGWSA-N 0.000 description 1
- KCTIFOCXAIUQQK-QXEWZRGKSA-N Ile-Pro-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O KCTIFOCXAIUQQK-QXEWZRGKSA-N 0.000 description 1
- LJHGALIOHLRRQN-DCAQKATOSA-N Leu-Ala-Arg Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CCCN=C(N)N LJHGALIOHLRRQN-DCAQKATOSA-N 0.000 description 1
- HASRFYOMVPJRPU-SRVKXCTJSA-N Leu-Arg-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O HASRFYOMVPJRPU-SRVKXCTJSA-N 0.000 description 1
- IASQBRJGRVXNJI-YUMQZZPRSA-N Leu-Cys-Gly Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)NCC(O)=O IASQBRJGRVXNJI-YUMQZZPRSA-N 0.000 description 1
- PPBKJAQJAUHZKX-SRVKXCTJSA-N Leu-Cys-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CS)C(=O)N[C@H](C(O)=O)CC(C)C PPBKJAQJAUHZKX-SRVKXCTJSA-N 0.000 description 1
- WIDZHJTYKYBLSR-DCAQKATOSA-N Leu-Glu-Glu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O WIDZHJTYKYBLSR-DCAQKATOSA-N 0.000 description 1
- YWYQSLOTVIRCFE-SRVKXCTJSA-N Leu-His-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(O)=O)C(O)=O YWYQSLOTVIRCFE-SRVKXCTJSA-N 0.000 description 1
- HRTRLSRYZZKPCO-BJDJZHNGSA-N Leu-Ile-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O HRTRLSRYZZKPCO-BJDJZHNGSA-N 0.000 description 1
- LXKNSJLSGPNHSK-KKUMJFAQSA-N Leu-Leu-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N LXKNSJLSGPNHSK-KKUMJFAQSA-N 0.000 description 1
- XVZCXCTYGHPNEM-UHFFFAOYSA-N Leu-Leu-Pro Natural products CC(C)CC(N)C(=O)NC(CC(C)C)C(=O)N1CCCC1C(O)=O XVZCXCTYGHPNEM-UHFFFAOYSA-N 0.000 description 1
- WMIOEVKKYIMVKI-DCAQKATOSA-N Leu-Pro-Ala Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O WMIOEVKKYIMVKI-DCAQKATOSA-N 0.000 description 1
- JIHDFWWRYHSAQB-GUBZILKMSA-N Leu-Ser-Glu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O JIHDFWWRYHSAQB-GUBZILKMSA-N 0.000 description 1
- SVBJIZVVYJYGLA-DCAQKATOSA-N Leu-Ser-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O SVBJIZVVYJYGLA-DCAQKATOSA-N 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
- 206010033266 Ovarian Hyperstimulation Syndrome Diseases 0.000 description 1
- UNLYPPYNDXHGDG-IHRRRGAJSA-N Phe-Gln-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 UNLYPPYNDXHGDG-IHRRRGAJSA-N 0.000 description 1
- CZQZSMJXFGGBHM-KKUMJFAQSA-N Phe-Pro-Gln Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CC2=CC=CC=C2)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O CZQZSMJXFGGBHM-KKUMJFAQSA-N 0.000 description 1
- KIZQGKLMXKGDIV-BQBZGAKWSA-N Pro-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1 KIZQGKLMXKGDIV-BQBZGAKWSA-N 0.000 description 1
- HFZNNDWPHBRNPV-KZVJFYERSA-N Pro-Ala-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O HFZNNDWPHBRNPV-KZVJFYERSA-N 0.000 description 1
- OYEUSRAZOGIDBY-JYJNAYRXSA-N Pro-Arg-Tyr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OYEUSRAZOGIDBY-JYJNAYRXSA-N 0.000 description 1
- UTAUEDINXUMHLG-FXQIFTODSA-N Pro-Asp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@@H]1CCCN1 UTAUEDINXUMHLG-FXQIFTODSA-N 0.000 description 1
- FKKHDBFNOLCYQM-FXQIFTODSA-N Pro-Cys-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CS)C(=O)N[C@@H](C)C(O)=O FKKHDBFNOLCYQM-FXQIFTODSA-N 0.000 description 1
- DIFXZGPHVCIVSQ-CIUDSAMLSA-N Pro-Gln-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(O)=O DIFXZGPHVCIVSQ-CIUDSAMLSA-N 0.000 description 1
- CLNJSLSHKJECME-BQBZGAKWSA-N Pro-Gly-Ala Chemical compound OC(=O)[C@H](C)NC(=O)CNC(=O)[C@@H]1CCCN1 CLNJSLSHKJECME-BQBZGAKWSA-N 0.000 description 1
- ZVEQWRWMRFIVSD-HRCADAONSA-N Pro-Phe-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=CC=C2)C(=O)N3CCC[C@@H]3C(=O)O ZVEQWRWMRFIVSD-HRCADAONSA-N 0.000 description 1
- JLMZKEQFMVORMA-SRVKXCTJSA-N Pro-Pro-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 JLMZKEQFMVORMA-SRVKXCTJSA-N 0.000 description 1
- HWLKHNDRXWTFTN-GUBZILKMSA-N Pro-Pro-Cys Chemical compound C1C[C@H](NC1)C(=O)N2CCC[C@H]2C(=O)N[C@@H](CS)C(=O)O HWLKHNDRXWTFTN-GUBZILKMSA-N 0.000 description 1
- RFWXYTJSVDUBBZ-DCAQKATOSA-N Pro-Pro-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 RFWXYTJSVDUBBZ-DCAQKATOSA-N 0.000 description 1
- AIOWVDNPESPXRB-YTWAJWBKSA-N Pro-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@@H]2CCCN2)O AIOWVDNPESPXRB-YTWAJWBKSA-N 0.000 description 1
- NBDHWLZEMKSVHH-UVBJJODRSA-N Pro-Trp-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@@H]3CCCN3 NBDHWLZEMKSVHH-UVBJJODRSA-N 0.000 description 1
- DWUIECHTAMYEFL-XVYDVKMFSA-N Ser-Ala-His Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 DWUIECHTAMYEFL-XVYDVKMFSA-N 0.000 description 1
- WDXYVIIVDIDOSX-DCAQKATOSA-N Ser-Arg-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CCCN=C(N)N WDXYVIIVDIDOSX-DCAQKATOSA-N 0.000 description 1
- KCGIREHVWRXNDH-GARJFASQSA-N Ser-Leu-Pro Chemical compound CC(C)C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N KCGIREHVWRXNDH-GARJFASQSA-N 0.000 description 1
- 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 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
- TYVAWPFQYFPSBR-BFHQHQDPSA-N Thr-Ala-Gly Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)NCC(O)=O TYVAWPFQYFPSBR-BFHQHQDPSA-N 0.000 description 1
- BSNZTJXVDOINSR-JXUBOQSCSA-N Thr-Ala-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(O)=O BSNZTJXVDOINSR-JXUBOQSCSA-N 0.000 description 1
- DGDCHPCRMWEOJR-FQPOAREZSA-N Thr-Ala-Tyr Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 DGDCHPCRMWEOJR-FQPOAREZSA-N 0.000 description 1
- NIEWSKWFURSECR-FOHZUACHSA-N Thr-Gly-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O NIEWSKWFURSECR-FOHZUACHSA-N 0.000 description 1
- FWTFAZKJORVTIR-VZFHVOOUSA-N Thr-Ser-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C)C(O)=O FWTFAZKJORVTIR-VZFHVOOUSA-N 0.000 description 1
- KAJRRNHOVMZYBL-IRIUXVKKSA-N Thr-Tyr-Gln 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](CCC(N)=O)C(O)=O KAJRRNHOVMZYBL-IRIUXVKKSA-N 0.000 description 1
- 102000004887 Transforming Growth Factor beta Human genes 0.000 description 1
- 108090001012 Transforming Growth Factor beta Proteins 0.000 description 1
- QUIXRGCMQOXUSV-SZMVWBNQSA-N Trp-Pro-Pro Chemical compound O=C([C@@H]1CCCN1C(=O)[C@H](CC=1C2=CC=CC=C2NC=1)N)N1CCC[C@H]1C(O)=O QUIXRGCMQOXUSV-SZMVWBNQSA-N 0.000 description 1
- CDKZJGMPZHPAJC-ULQDDVLXSA-N Tyr-Leu-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 CDKZJGMPZHPAJC-ULQDDVLXSA-N 0.000 description 1
- SZEIFUXUTBBQFQ-STQMWFEESA-N Tyr-Pro-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O SZEIFUXUTBBQFQ-STQMWFEESA-N 0.000 description 1
- ASQFIHTXXMFENG-XPUUQOCRSA-N Val-Ala-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)NCC(O)=O ASQFIHTXXMFENG-XPUUQOCRSA-N 0.000 description 1
- XQVRMLRMTAGSFJ-QXEWZRGKSA-N Val-Asp-Arg Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N XQVRMLRMTAGSFJ-QXEWZRGKSA-N 0.000 description 1
- RKIGNDAHUOOIMJ-BQFCYCMXSA-N Val-Glu-Trp Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)C(C)C)C(O)=O)=CNC2=C1 RKIGNDAHUOOIMJ-BQFCYCMXSA-N 0.000 description 1
- KZKMBGXCNLPYKD-YEPSODPASA-N Val-Gly-Thr Chemical compound CC(C)[C@H](N)C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(O)=O KZKMBGXCNLPYKD-YEPSODPASA-N 0.000 description 1
- XTDDIVQWDXMRJL-IHRRRGAJSA-N Val-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C(C)C)N XTDDIVQWDXMRJL-IHRRRGAJSA-N 0.000 description 1
- UEXPMFIAZZHEAD-HSHDSVGOSA-N Val-Thr-Trp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](C(C)C)N)O UEXPMFIAZZHEAD-HSHDSVGOSA-N 0.000 description 1
- AEFJNECXZCODJM-UWVGGRQHSA-N Val-Val-Gly Chemical compound CC(C)[C@H]([NH3+])C(=O)N[C@@H](C(C)C)C(=O)NCC([O-])=O AEFJNECXZCODJM-UWVGGRQHSA-N 0.000 description 1
- 108010045350 alanyl-tyrosyl-alanine Proteins 0.000 description 1
- 108010070944 alanylhistidine Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 108010013835 arginine glutamate Proteins 0.000 description 1
- 108010069926 arginyl-glycyl-serine Proteins 0.000 description 1
- 108010029539 arginyl-prolyl-proline Proteins 0.000 description 1
- 108010060035 arginylproline Proteins 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 229940098773 bovine serum albumin Drugs 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 230000011712 cell development Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 238000003759 clinical diagnosis Methods 0.000 description 1
- 239000011162 core material Substances 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 108010004073 cysteinylcysteine Proteins 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000035558 fertility Effects 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 108010006664 gamma-glutamyl-glycyl-glycine Proteins 0.000 description 1
- 108010055341 glutamyl-glutamic acid Proteins 0.000 description 1
- JYPCXBJRLBHWME-UHFFFAOYSA-N glycyl-L-prolyl-L-arginine Natural products NCC(=O)N1CCCC1C(=O)NC(CCCN=C(N)N)C(O)=O JYPCXBJRLBHWME-UHFFFAOYSA-N 0.000 description 1
- 108010000434 glycyl-alanyl-leucine Proteins 0.000 description 1
- 108010019832 glycyl-asparaginyl-glycine Proteins 0.000 description 1
- 108010084264 glycyl-glycyl-cysteine Proteins 0.000 description 1
- 108010079413 glycyl-prolyl-glutamic acid Proteins 0.000 description 1
- 108010077515 glycylproline Proteins 0.000 description 1
- 210000002503 granulosa cell Anatomy 0.000 description 1
- 239000005337 ground glass Substances 0.000 description 1
- 108010025306 histidylleucine Proteins 0.000 description 1
- 239000000710 homodimer Substances 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000009245 menopause Effects 0.000 description 1
- 239000011259 mixed solution Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 239000000178 monomer Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 210000001672 ovary Anatomy 0.000 description 1
- 238000011056 performance test Methods 0.000 description 1
- RXNXLAHQOVLMIE-UHFFFAOYSA-N phenyl 10-methylacridin-10-ium-9-carboxylate Chemical compound C12=CC=CC=C2[N+](C)=C2C=CC=CC2=C1C(=O)OC1=CC=CC=C1 RXNXLAHQOVLMIE-UHFFFAOYSA-N 0.000 description 1
- 108010077112 prolyl-proline Proteins 0.000 description 1
- 108010093296 prolyl-prolyl-alanine Proteins 0.000 description 1
- 108010031719 prolyl-serine Proteins 0.000 description 1
- 108010029020 prolylglycine Proteins 0.000 description 1
- 238000001959 radiotherapy Methods 0.000 description 1
- 230000009257 reactivity Effects 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000000717 sertoli cell Anatomy 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 210000004989 spleen cell Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 210000001550 testis Anatomy 0.000 description 1
- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 1
- 108010084932 tryptophyl-proline Proteins 0.000 description 1
Images
Classifications
-
- 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
- G01N33/74—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors
-
- 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
- C07K16/26—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against hormones ; against hormone releasing or inhibiting factors
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
-
- 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
- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
- G01N33/54313—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being characterised by its particulate form
- G01N33/54326—Magnetic particles
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2317/00—Immunoglobulins specific features
- C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
-
- 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
- G01N2333/475—Assays involving growth factors
- G01N2333/495—Transforming growth factor [TGF]
-
- 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
- G01N2333/575—Hormones
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Urology & Nephrology (AREA)
- Hematology (AREA)
- Biomedical Technology (AREA)
- General Health & Medical Sciences (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Analytical Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Pathology (AREA)
- General Physics & Mathematics (AREA)
- Cell Biology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Food Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Organic Chemistry (AREA)
- Endocrinology (AREA)
- Plasma & Fusion (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides an anti-mullerian hormone detection kit, a monoclonal antibody and a hybridoma cell strain, wherein the kit comprises a first monoclonal antibody and a second monoclonal antibody; the first monoclonal antibody is produced by a hybridoma cell strain with the preservation number of CGMCC NO. 23868; the second monoclonal antibody is produced by hybridoma cell strain with preservation number of CGMCC NO. 23869. The kit provided by the invention can obtain a stable detection result.
Description
Technical Field
The invention relates to the technical field of immunodetection, and particularly relates to an anti-mullerian hormone detection kit, a monoclonal antibody and a hybridoma cell strain.
Background
Anti-mullerian hormone (AMH), one of the members of the transforming growth factor β superfamily, was first discovered by Alfred Jost in 1947 because of its role in regulating cell development and differentiation, causing the degeneration of male embryonic mullerian tubes. Anti-mullerian hormone is a glycoprotein secreted by Sertoli cells of immature testes and granulosa cells of the ovarian antral and antral follicles. The greater the number of antral follicles in the ovary, the higher the concentration of AMH; on the contrary, as the follicles are gradually depleted with age and various factors, the AMH concentration decreases, and as the menopause approaches, AMH gradually approaches 0, which can be used as a diagnostic marker for predicting ovarian reserve. The AMH is indispensable in the auxiliary reproduction technical process, can predict ovarian reactivity, identifies women at risk of ovarian hyperstimulation syndrome, can judge the dosage of ovulation-promoting drugs according to the AMH value, and improves the auxiliary reproduction success rate. The clinical significance of AMH also includes prediction of Premature Ovarian Failure (POF), menopausal age, auxiliary diagnosis of polycystic ovarian syndrome (PCOS), monitoring of damage and recovery of ovarian function caused by operation, radiotherapy and chemotherapy, and the like. The anti-mullerian hormone (AMH) is used as the optimal index for measuring the ovarian function, has increasingly prominent important function in the evaluation of female fertility and the auxiliary reproduction, and has very large market application prospect.
The AMH detection method commonly used in clinic at present mainly comprises enzyme-linked immunoassay, chemiluminescence immunoassay and the like. These techniques are based on the specific interaction of antigen and antibody, the specific antibody being the core material of these detection methods. However, the AMH index has a phenomenon that the measured value is unstable, and unlike most indexes that are unstable in that the measured value decreases, the same sample increases the AMH measured value as the storage time increases. The rise amplitude of the sample is related to the sample preservation temperature, time and preservation mode (serum or whole blood), and the rise amplitude of the sample is different from person to person.
Disclosure of Invention
In order to solve the problems, the invention provides an anti-mullerian hormone immunoassay kit, a monoclonal antibody and a hybridoma cell strain. The kit provided by the invention can obtain a stable detection result.
The invention provides an anti-mullerian hormone immunoassay kit, which comprises a first monoclonal antibody and a second monoclonal antibody; wherein the first monoclonal antibody is produced by a hybridoma cell strain with the preservation number of CGMCC NO. 23868; the second monoclonal antibody is produced by hybridoma cell strain with preservation number of CGMCC NO. 23869.
Further, the binding site of the first monoclonal antibody and anti-mullerian hormone is located in the sequence shown in SEQ ID No. 19. Further, the binding sites of the first monoclonal antibody and the anti-mullerian hormone are: glu Thr Tyr Gln Ala Asn Asn Cys Gln Gly Val Cys Gly Trp Pro Gln Ser Asp Arg Asn are provided.
Further, the binding site of the second monoclonal antibody and anti-mullerian hormone is located in the sequence shown in SEQ ID No. 17. Further, the binding sites for the second mab and anti-mullerian hormone are: leu Gln Gly Leu Arg Val Glu Trp Arg Gly Arg Asp Pro Arg Gly Pro Gly Arg Ala Gln are provided.
Further, the kit comprises a capture antibody and a detection antibody; wherein, the capture antibody is crosslinked with magnetic beads, and the detection antibody is crosslinked with luminescent markers; when either of the first monoclonal antibody and the second monoclonal antibody is a capture antibody, the other is a detection antibody.
In another aspect, embodiments of the present invention also provide a monoclonal antibody, comprising: a first monoclonal antibody produced by a hybridoma cell strain with the preservation number of CGMCC NO. 23868; or a second monoclonal antibody produced by a hybridoma cell strain with the preservation number of CGMCC NO. 23869.
Further, the monoclonal antibody is obtained by screening polypeptide; the polypeptide is selected from the following sequences: SEQ ID NO.1 to SEQ ID NO. 21.
Further, the monoclonal antibody is used for preparing an immunoassay tool for detecting the anti-mullerian hormone.
On the other hand, the invention also provides a hybridoma cell strain, wherein the preservation number of the hybridoma cell strain is CGMCC NO. 23868; or the preservation number of the hybridoma cell strain is CGMCC NO. 23869.
Compared with the prior art, the technical scheme provided by the invention has the beneficial effects that:
1. the immunoassay kit provided by the invention has the advantages of stable AMH measured value, strong specificity and high sensitivity.
2. The hybrid polypeptide is adopted to screen hybridoma cell strains to obtain specific monoclonal antibodies, and meanwhile, the hybrid polypeptide can also detect the binding sites of the monoclonal antibodies, so that the problems that the conformation of AMH molecules in a sample is easy to change and the monoclonal antibodies are sensitive to the binding sites are effectively solved.
3. The screening process of the monoclonal antibody is simple, and meanwhile, the binding site of the monoclonal antibody is a linear epitope which is different from the existing site.
4. The monoclonal antibody provided by the invention can be used for detecting anti-mullerian hormone (AMH) by a chemiluminescence method.
Biological preservation description:
the classification of hybridoma cell line 12D7B6 for preservation was designated:
the preservation unit is fully called as follows: china general microbiological culture Collection center;
the preservation unit is abbreviated as: CGMCC;
the address of the depository: xilu No.1 Hospital No. 3, Beijing, Chaoyang, North;
the preservation date is as follows: 11/12/2021;
the preservation number is: CGMCC NO. 23868.
The classification of hybridoma cell line 5A3H9 for preservation was named:
the preservation unit is called as follows: china general microbiological culture Collection center;
the preservation unit is abbreviated as: CGMCC;
the address of the depository: xilu No.1 Hospital No. 3, Beijing, Chaoyang, Beicheng;
the preservation date is as follows: 11/12/2021;
the preservation number is: CGMCC number 23869.
Drawings
FIG. 1 is a linear fit curve of the correlation between the kit and Roche reagent provided by the embodiment of the invention.
Detailed Description
In order to make the aforementioned objects, features and advantages of the present invention comprehensible, embodiments accompanied with figures are described in detail below to clearly and completely describe the technical solutions in the embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The AMH protein full length 560AA is secreted extracellularly in the form of disulfide-linked homodimer, each monomer can be cleaved from 451AA/452AA into N terminal (AMHN) and "mature region" C terminal (AMHC), but the N terminal and the C terminal of the cleaved dimer are not separated and are combined together in a non-covalent bond manner to form 140kD glycoprotein consisting of 4 peptide chains. The human serum contains the same amount of cleaved and uncleaved dimeric protein, and the ratio of cleaved and uncleaved dimeric protein is different in different human serums.
The phenomenon of unstable measured value of the AMH index exists at present, which is related to the dimer molecular structure of the AMH, and the molecular conception of a sample is changed in the preservation process, so that a new body binding site is exposed, and the measured value is increased. Different antibody binding sites differ, and antigen-antibody binding is affected differently by changes in antigen conformation, so that stable, reproducible AMH marker detection results in clinical diagnosis rely on antibodies that are not susceptible to changes in antigen conformation. None of the commonly used methods for antibody preparation address the problem of measured value stability of the sample. However, the preparation process of the monoclonal antibody of the method is complex, and it is not proved that all the monoclonal antibody pairs meeting the conditions can be used for AMH detection and can obtain a stable detection result, and meanwhile, the antibody of the non-site can not obtain a stable AMH detection result.
Aiming at the problems, the invention provides a hybridoma cell strain 12D7B6 which is preserved in China general microbiological culture Collection center (CGMCC for short) with the preservation date of CGMCC number 23868; the hybridoma cell strain 5A3H9 is preserved in China general microbiological culture Collection center (CGMCC for short) with the preservation date of CGMCC number 23869;
the invention also provides a monoclonal antibody, which comprises a first monoclonal antibody and a second monoclonal antibody; wherein the first monoclonal antibody is secreted by a hybridoma cell strain 12D7B6, and the second monoclonal antibody is secreted by a hybridoma cell strain 5A3H 9.
Further, the monoclonal antibody is used for preparing an immunoassay tool for detecting the anti-mullerian hormone.
In the present embodiment, the immunoassay means, i.e. the means for detecting anti-mullerian hormone, is not specifically limited herein, and it is within the scope of the present invention to refer to the means well known to those skilled in the art, for example, the immunoassay means can be a reagent, a kit, a test strip, a chip, and other conventional immunoassay means.
Further, the monoclonal antibody was prepared as follows:
(1) preparation of immunogen: synthesizing an AMH full-length (1-560 AA) gene sequence, adding an 8 XHis tag at the C terminal, and inserting pcDNA3.3 plasmid into the C terminal by using a 5'NheI and 3' EcoRI enzyme cutting site to construct an expression vector. 293T cells are clockwise transferred by adopting a conventional technical method, and the AMH full-length antigen is obtained by affinity purification through a nickel column.
(2) Animal immunization: the AMH full-length antigen is adopted to immunize a Balb/c mouse, immune spleen cells of the mouse are taken to be fused with SP2/0 cells, and positive hybridoma cell strains are screened by indirect ELISA to obtain hybridoma cell strains 12D7B6 and hybridoma cell strains 5A3H 9. And performing subcloning on the two hybridoma cell strains, culturing to prepare an antibody, and purifying to obtain a first monoclonal antibody and a second monoclonal antibody.
(3) Antibody pairing: and adding an AMH full-length antigen into the first monoclonal antibody, carrying out immunoreaction, adding a second antibody after cleaning, adding a color developing agent after cleaning again, and successfully pairing after color development. Similarly, the antibody can also be paired by adding AMH full-length antigen into the second monoclonal antibody, carrying out immunoreaction, adding the first antibody after cleaning, adding the color developing agent after cleaning again, and then successfully pairing after color development.
Further, hybridoma cell strains are obtained by polypeptide screening; the polypeptide is selected from the following sequences: SEQ ID NO.1 to SEQ ID NO. 21.
In this embodiment, the screening of positive hybridoma cell lines by indirect ELISA is aimed at screening hybridoma cell lines that produce specific antibodies, and in some embodiments of the present invention, the screening is performed using a mixed polypeptide, which specifically includes the following steps:
(1) synthesizing a polypeptide: synthesizing polypeptides shown as SEQ ID NO. 1-SEQ ID NO.21 sequences, correspondingly obtaining polypeptides No. 1-21, wherein each polypeptide sequence has 30AA, and 5AA of adjacent polypeptides are mutually overlapped; wherein the polypeptides No. 1-17 are located at AMHN, and the polypeptides No. 18-21 are located at AMHC.
(2) Preparation of mixed polypeptide: mixing the polypeptides No. 1-4 in equal proportion, and coupling BSA; mixing the polypeptides No. 5-8 in equal proportion, and coupling BSA; mixing the 9-13 polypeptides in equal proportion, and coupling BSA; mixing 14-17 polypeptide in equal proportion, and coupling BSA; the polypeptides No. 18-21 are mixed in equal proportion and coupled with BSA. And (5) subpackaging and storing.
(3) The mixed polypeptide is adopted to detect positive hybridoma cells by indirect ELISA.
The embodiment of the invention also provides an anti-mullerian hormone immunoassay kit, which comprises the first monoclonal antibody and the second monoclonal antibody.
Further, the kit comprises a capture antibody and a detection antibody; wherein, the capture antibody is crosslinked with magnetic beads, and the detection antibody is crosslinked with luminescent markers; when either of the first monoclonal antibody and the second monoclonal antibody is a capture antibody, the other is a detection antibody.
In this example, the preparation of the kit comprises the following steps:
(1) preparation of a luminescent reagent: and (3) labeling the detection antibody by using acridine, sequentially carrying out light-resistant reaction and dialysis desalination, and finally diluting and storing for later use.
(2) Preparation of solid-phase reagents: and (3) labeling a capture antibody by using biotin, labeling the biotin by using magnetic beads after dialysis and desalination, washing, diluting and storing for later use.
Further, the binding site of the first monoclonal antibody and anti-mullerian hormone is located in the sequence shown in SEQ ID No. 19. Further, the binding sites of the first monoclonal antibody and the anti-mullerian hormone are: glu Thr Tyr Gln Ala Asn Asn Cys Gln Gly Val Cys Gly Trp Pro Gln Ser Asp Arg Asn is added.
Further, the binding site of the second monoclonal antibody and anti-mullerian hormone is located in the sequence shown in SEQ ID No. 17. Further, the binding sites for the second mab and anti-mullerian hormone are: leu Gln Gly Leu Arg Val Glu Trp Arg Gly Arg Asp Pro Arg Gly Pro Gly Arg Ala Gln are provided.
The technical solution provided by the present invention will be further described with reference to specific embodiments.
Example 1
Preparation of monoclonal antibody
1. Preparation of immunogens
Synthesizing an AMH full-length (1-560 AA) gene sequence, adding an 8 XHis tag at the C terminal, and inserting pcDNA3.3 plasmid into the C terminal by using a 5'NheI and 3' EcoRI enzyme cutting site to construct an expression vector. 293T cells are clockwise transferred by adopting a conventional technical method, and the AMH full-length antigen is obtained by affinity purification through a nickel column.
2. Preparation of Mixed polypeptide-BSA conjugates
Synthesizing polypeptides shown as SEQ ID NO. 1-SEQ ID NO.21 sequences, correspondingly obtaining polypeptides No. 1-21, wherein each polypeptide sequence has 30AA, and 5AA of adjacent polypeptides are mutually overlapped; wherein the polypeptide No. 1-17 is located in AMHN, and the polypeptide No. 18-21 is located in AMHC.
Weighing 2mg BSA and dissolving in 600ul PBS pH7.4; weighing 0.5mg of sulf-SMCC and dissolving in 100ul of PBS; then adding the dissolved SMCC solution into a BSA solution, and then reacting at room temperature for 0.5 hour; the solution was added to the dialysis bag and dialyzed against PBS pH7.4 to remove excess SMCC and the dialysate was changed 3 times. And (3) mixing the polypeptides No. 1-4 in equal proportion, slowly dripping 1mg of the mixture into activated BSA while uniformly mixing, reacting at room temperature for 2 hours, and measuring the concentration by using a BCA method.
Similarly, polypeptides 5-8 are mixed in equal proportion and coupled with BSA; mixing the 9-13 polypeptides in equal proportion, and coupling BSA; mixing 14-17 polypeptide in equal proportion, and coupling BSA; the polypeptides No. 18-21 are mixed in equal proportion and coupled with BSA. And (5) subpackaging and storing.
3. Animal immunization
Selecting female Balb/c mice with strong bodies of 6-8 weeks of age, and immunizing with the prepared AMH full-length antigen. The first immunization adopts Freund's complete adjuvant, 100 mug/mouse and multi-point injection under the armpit; boosting, adopting Freund incomplete adjuvant, 50 mug/mouse, boosting 3 times, the boosting interval time of adjacent times is 2 weeks; the first immunization was 3 weeks apart from the first booster immunization. Blood is collected by eyeballs 1 week after the last boosting immunization, an enzyme label plate is coated by AMH full-length antigen, and the titer of antiserum is measured by an indirect ELISA method. 3 days before fusion, 100. mu.g antigen/mouse was injected intraperitoneally with a shock.
The impacted mice were sacrificed by cervical dislocation and sterilized by immersion in 75% ethanol. In a clean bench, the spleens of mice were aseptically removed, squeezed with a sterilized ground glass slide, and lightly ground to isolate splenocytes, which were counted. The SP2/0 cells and splenocytes were mixed at a ratio of 1:1, centrifuged at 300g for 5 minutes at room temperature, and the supernatant was discarded. 1ml of PEG was added and the PEG and cells were mixed well and homogeneously, this was completed within 3 minutes. 50ml of fresh DMEM medium is added, centrifuged at room temperature 300g for 5 minutes, and HAT medium is resuspended and gently mixed. 100 ul/well was added to a 96-well plate and placed in an incubator for culture.
HT medium was changed when cells grew to 1/3 basal area in 96-well plates, mixed polypeptide-BSA conjugates were coated at 2ug/ml, and positive clones were detected by indirect ELISA. And performing subcloning for 3-4 times to ensure that the amplification culture is performed when the monoclonal antibody is formed, performing ascites cell injection, and purifying to obtain the first monoclonal antibody and the second monoclonal antibody.
Second, antibody labeling and pairing
1. Antibody labeling
The first monoclonal antibody and the second monoclonal antibody were diluted to a concentration of 1mg/ml with 1 xPBS, and 5ul of Biotin (30 mg/ml, EZ-Link NHS-PEG4-Biotin, Thermo) was added to 2mg of antibody and mixed rapidly. After mixing, the centrifuge tube with the mixed solution is placed on a vertical stirrer to react for 30min or overnight at 4 ℃. Biotinylated antibodies were dialyzed against 1 xPBS 3 times with changes every 4 hours. Obtaining the first monoclonal antibody marked by biotin and the second monoclonal antibody marked by biotin.
2. Antibody pairing
Incubating the first monoclonal antibody at 37 ℃ for 1h for coating, wherein the concentration of the first monoclonal antibody in each well is 2 mu g/mlx 100 mu l; blocking with 200. mu.L of 2% BSA, incubation at 37 ℃ for 1h, and washing with PBST 3 times. 5ng/ml of the prepared AMH full-length antigen was added to each well, a negative control without antigen was set for each pair, incubated at 37 ℃ for 1h, and washed 3 times with PBST. Adding biotin-labeled second monoclonal antibody (1: 200), incubating at 37 ℃ for 1h, and washing with PBST for 3 times; adding SA-HRP 1:2000, 100 μ l, incubating at 37 deg.C for 30min, and washing with PBST for 4 times; color development, reading OD value at 450nm wavelength. And OD is 2.1 times larger than that of the negative control hole, positive pairing is judged, and the antibody with high signal-to-noise ratio is selected for preparing a chemiluminescence reagent for sample detection stability test.
Incubating the second monoclonal antibody at 37 ℃ for 1h for coating, wherein the second monoclonal antibody is 2 mu g/mlx 100 mu l per well; blocking with 200. mu.L of 2% BSA, incubation at 37 ℃ for 1h, and washing with PBST 3 times. 5ng/ml of the prepared AMH full-length antigen was added to each well, a negative control without antigen was set for each pair, incubated at 37 ℃ for 1h, and washed 3 times with PBST. Adding biotin-labeled first monoclonal antibody (1: 200), incubating at 37 ℃ for 1h, and washing with PBST for 3 times; adding SA-HRP 1:2000, 100 μ l, incubating at 37 deg.C for 30min, and washing with PBST for 4 times; color development, reading OD value at 450nm wavelength. And OD is 2.1 times larger than that of the negative control hole, positive pairing is judged, and the antibody with high signal-to-noise ratio is selected for preparing a chemiluminescence reagent for sample detection stability test.
Preparation of immunoassay kit
1. Preparation of luminescent reagents
Taking 0.1mg of detection antibody, adding acridinium ester label according to the molar ratio of 1:5-1:15, uniformly mixing at room temperature in the dark for reaction for 2h, dialyzing and desalting the PB buffer solution in the dark, wherein the volume of the dialysate is 20 times larger than that of the label, changing the dialysate once every 4 hours, changing the dialysate for at least 3 times, and diluting the dialysate to 1-10 mu g/mL by PBS-BSA after measuring the concentration for later use.
2. Solid phase reagent preparation
Taking 0.1mg of capture antibody, adding biotin label in a molar ratio of 1:5-1:15, uniformly mixing at room temperature, reacting for 2h, dialyzing and desalting with 20mM PB buffer solution, wherein the volume of dialysate is 20 times larger than that of the label, changing the dialysate once every 4 hours, changing the dialysate at least 3 times, and diluting with PBS-BSA to 0.1-1mg/mL for later use after measuring the concentration.
And washing PB (magnetic bead) by about 0.5mL for three times, diluting to 1-10mg/mL, adding a biotin labeled antibody in an equal volume, uniformly mixing at room temperature for 2h, washing with PBS-BSA for three times, and diluting to 0.1-1mg/mL for later use.
When either of the first monoclonal antibody and the second monoclonal antibody is a capture antibody, the other is a detection antibody.
3. Sample detection stability test
Fresh serum of women aged more than 55 years (AMH negative) and less than 50 years (AMH positive) is collected respectively, part of the samples are used for preparing negative mixed samples (5 serum samples), positive mixed samples are mixed with 1 (5 serum samples) and mixed with 2 (5 serum samples). The test result of the serum sample collection on the day is marked as the 0 th day, all samples are subpackaged and stored at the temperature of 2-8 ℃ and the room temperature (about 25 ℃) after the test is finished, the samples are taken out for retesting after 3 days, and the change ratio is calculated by dividing the test result by the test result of the 0 th day. Mixing the negative sample with positive samples mixed 1 and mixed 2 was controlled by simultaneous testing with another pair of antibodies known to have elevated values. And (3) testing conditions are as follows: meikang biological production chemiluminescence immunoassay analyzer MS120, one-step method, 100u L luminescence reagent, 50 u L solid phase reagent, 30 u L sample, 37 degrees C were incubated for 15 min.
As shown in Table 1, the 12D7B6 clone (capture) and the 5A3H9 clone (detection) were paired and tested on single serum samples and mixed samples stored at 2-8 ℃ or 25 ℃ for 3 days, the brightness value of the single serum samples and the mixed samples did not increase significantly relative to the test result of 0 day, while the brightness value of the control antibody of the single serum samples and the mixed samples stored at 3 days was increased significantly relative to the test result of 1 and 2 samples, and the measured value of the samples stored at 25 ℃ increased by more than 2-8 ℃.
TABLE 1
Fourthly, performance test of the kit
1. Correlation test
50 clinical serum samples were collected and the theoretical values covered the linear range of 0.01-20ng/ml, and tested using the Roche Elecsys AMH diagnostic kit and the electrochemiluminescence analyzer under the conditions described in the specification. The same positive sample is detected by the self-prepared luminescent reagent and the solid phase reagent, and the test conditions are as follows: one step, 100. mu.L of luminescent reagent, 50. mu.L of solid phase reagent, 30. mu.L of sample, incubated at 37 ℃ for 15 min. Recording the test result, and calculating the linear fitting correlation curve equation and correlation coefficient R of the test result of the self-prepared reagent and the Roche reagent2. Referring to fig. 1 and table 2, a linear fit of the test results of the luminescent reagent prepared by pairing clone 12D7B6 (capture) with clone 5A3H9 (detection) to the measurement of the Elecsys AMH diagnostic kit is shown as: y = 83073x-2864, R = 0.9902, self-prepared luminescent reagent has better correlation with the measured value of the Elecsys AMH diagnostic kit.
TABLE 2
2. Lowest detection line
Taking 5% bovine serum as a blank sample, continuously measuring for 25 times, and testing conditions are as follows: one-step method, 100. mu.L luminescence reagent, 50. mu.L solid phase reagent, 30. mu.L sample (5% bovine serum blank sample), and incubation at 37 ℃ for 15 min. The test results were recorded, and the average value av, the standard deviation sd, and the lowest detection line LOB ═ av +2sd were calculated, respectively, as shown in table 3, and the lowest detection line for the reagent prepared by pairing 5A3H9 with 12D7B6 was less than 0.1 ng/ml.
TABLE 3
Fifth, antibody binding site detection
Coating a 96-well plate with polypeptide-BSA conjugates 1-21 at a concentration of 1. mu.g/ml; 2% bovine serum albumin, 1 small test at 37 ℃, PBS pH7.4 washing three times; diluting the monoclonal antibody to 1 mu g/ml, adding the diluted monoclonal antibody into 100 mu l/well, reacting at 37 ℃ for 1 hour, and washing with PBS (phosphate buffer solution) pH7.4 for three times; adding a goat anti-mouse secondary antibody diluted at a ratio of 1:2000, reacting at a concentration of 100. mu.l/well for 1 hour at 37 ℃, and washing with PBS (phosphate buffer solution) pH7.4 for four times; TMB was developed for 5 minutes, and the reaction was terminated. The data are read and analyzed, and the antibody binding sites are determined based on the positively reacting polypeptide sequences.
As shown in Table 4, the 5A3H9 clone bound to polypeptide No.17 but not to polypeptide Nos. 16 and 18, and the Amh-17 clone binding site was presumed to be located at 431AA-450 AA. The 12D7B6 clone could bind to polypeptide 19 but not to polypeptide 18 or 20, and the Amh-19 clone binding site was presumed to be located at 481AA-500 AA.
TABLE 4
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Sequence listing
<110> Meikang Biotechnology Ltd
<120> anti-mullerian hormone detection kit, monoclonal antibody and hybridoma cell strain
<130> 2022.02.21
<160> 21
<170> SIPOSequenceListing 1.0
<210> 1
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 1
Arg Ala Glu Glu Pro Ala Val Gly Thr Ser Gly Leu Ile Phe Arg Glu
1 5 10 15
Asp Leu Asp Trp Pro Pro Gly Ser Pro Gln Glu Pro Leu Cys Gly Gly
20 25 30
Cys
<210> 2
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 2
Gln Glu Pro Leu Cys Leu Val Ala Leu Gly Gly Asp Ser Asn Gly Ser
1 5 10 15
Ser Ser Pro Leu Arg Val Val Gly Ala Leu Ser Ala Tyr Glu Gly Gly
20 25 30
Cys
<210> 3
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 3
Leu Ser Ala Tyr Glu Gln Ala Phe Leu Gly Ala Val Gln Arg Ala Arg
1 5 10 15
Trp Gly Pro Arg Asp Leu Ala Thr Phe Gly Val Cys Asn Thr Gly Gly
20 25 30
Cys
<210> 4
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 4
Gly Val Cys Asn Thr Gly Asp Arg Gln Ala Ala Leu Pro Ser Leu Arg
1 5 10 15
Arg Leu Gly Ala Trp Leu Arg Asp Pro Gly Gly Gln Arg Leu Gly Gly
20 25 30
Cys
<210> 5
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 5
Gly Gly Gln Arg Leu Val Val Leu His Leu Glu Glu Val Thr Trp Glu
1 5 10 15
Pro Thr Pro Ser Leu Arg Phe Gln Glu Pro Pro Pro Gly Gly Gly Gly
20 25 30
Cys
<210> 6
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 6
Pro Pro Pro Gly Gly Ala Gly Pro Pro Glu Leu Ala Leu Leu Val Leu
1 5 10 15
Tyr Pro Gly Pro Gly Pro Glu Val Thr Val Thr Arg Ala Gly Gly Gly
20 25 30
Cys
<210> 7
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 7
Val Thr Arg Ala Gly Leu Pro Gly Ala Gln Ser Leu Cys Pro Ser Arg
1 5 10 15
Asp Thr Arg Tyr Leu Val Leu Ala Val Asp Arg Pro Ala Gly Gly Gly
20 25 30
Cys
<210> 8
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 8
Asp Arg Pro Ala Gly Ala Trp Arg Gly Ser Gly Leu Ala Leu Thr Leu
1 5 10 15
Gln Pro Arg Gly Glu Asp Ser Arg Leu Ser Thr Ala Arg Leu Gly Gly
20 25 30
Cys
<210> 9
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 9
Ser Thr Ala Arg Leu Gln Ala Leu Leu Phe Gly Asp Asp His Arg Cys
1 5 10 15
Phe Thr Arg Met Thr Pro Ala Leu Leu Leu Leu Pro Arg Ser Gly Gly
20 25 30
Cys
<210> 10
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 10
Leu Leu Pro Arg Ser Glu Pro Ala Pro Leu Pro Ala His Gly Gln Leu
1 5 10 15
Asp Thr Val Pro Phe Pro Pro Pro Arg Pro Ser Ala Glu Leu Gly Gly
20 25 30
Cys
<210> 11
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 11
Pro Ser Ala Glu Leu Glu Glu Ser Pro Pro Ser Ala Asp Pro Phe Leu
1 5 10 15
Glu Thr Leu Thr Arg Leu Val Arg Ala Leu Arg Val Pro Pro Gly Gly
20 25 30
Cys
<210> 12
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 12
Leu Arg Val Pro Pro Ala Arg Ala Ser Ala Pro Arg Leu Ala Leu Asp
1 5 10 15
Pro Asp Ala Leu Ala Gly Phe Pro Gln Gly Leu Val Asn Leu Gly Gly
20 25 30
Cys
<210> 13
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 13
Gly Leu Val Asn Leu Ser Asp Pro Ala Ala Leu Glu Arg Leu Leu Asp
1 5 10 15
Gly Glu Glu Pro Leu Leu Leu Leu Leu Arg Pro Thr Ala Ala Gly Gly
20 25 30
Cys
<210> 14
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 14
Arg Pro Thr Ala Ala Thr Thr Gly Asp Pro Ala Pro Leu His Asp Pro
1 5 10 15
Thr Ser Ala Pro Trp Ala Thr Ala Leu Ala Arg Arg Val Ala Gly Gly
20 25 30
Cys
<210> 15
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 15
Ala Arg Arg Val Ala Ala Glu Leu Gln Ala Ala Ala Ala Glu Leu Arg
1 5 10 15
Ser Leu Pro Gly Leu Pro Pro Ala Thr Ala Pro Leu Leu Ala Gly Gly
20 25 30
Cys
<210> 16
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 16
Ala Pro Leu Leu Ala Arg Leu Leu Ala Leu Cys Pro Gly Gly Pro Gly
1 5 10 15
Gly Leu Gly Asp Pro Leu Arg Ala Leu Leu Leu Leu Lys Ala Gly Gly
20 25 30
Cys
<210> 17
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 17
Leu Leu Leu Lys Ala Leu Gln Gly Leu Arg Val Glu Trp Arg Gly Arg
1 5 10 15
Asp Pro Arg Gly Pro Gly Arg Ala Gln Arg Ser Ala Gly Ala Gly Gly
20 25 30
Cys
<210> 18
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 18
Arg Ser Ala Gly Ala Thr Ala Ala Asp Gly Pro Cys Ala Leu Arg Glu
1 5 10 15
Leu Ser Val Asp Leu Arg Ala Glu Arg Ser Val Leu Ile Pro Gly Gly
20 25 30
Cys
<210> 19
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 19
Ser Val Leu Ile Pro Glu Thr Tyr Gln Ala Asn Asn Cys Gln Gly Val
1 5 10 15
Cys Gly Trp Pro Gln Ser Asp Arg Asn Pro Arg Tyr Gly Asn Gly Gly
20 25 30
Cys
<210> 20
<211> 33
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 20
Pro Arg Tyr Gly Asn His Val Val Leu Leu Leu Lys Met Gln Val Arg
1 5 10 15
Gly Ala Ala Leu Ala Arg Pro Pro Cys Cys Val Pro Thr Ala Gly Gly
20 25 30
Cys
<210> 21
<211> 38
<212> PRT
<213> Artificial Sequence (Artificial Sequence)
<400> 21
Cys Val Pro Thr Ala Tyr Ala Gly Lys Leu Leu Ile Ser Leu Ser Glu
1 5 10 15
Glu Arg Ile Ser Ala His His Val Pro Asn Met Val Ala Thr Glu Cys
20 25 30
Gly Cys Arg Gly Gly Cys
35
Claims (10)
1. An anti-mullerian hormone immunoassay kit is characterized in that,
comprises a first monoclonal antibody and a second monoclonal antibody;
wherein the first monoclonal antibody is produced by a hybridoma cell strain with the preservation number of CGMCC NO. 23868;
the second monoclonal antibody is produced by a hybridoma cell strain with the preservation number of CGMCC NO. 23869.
2. The kit according to claim 1,
the binding site of the first monoclonal antibody and the anti-mullerian hormone is positioned in a sequence shown in SEQ ID NO. 19.
3. The kit according to claim 2,
the binding sites of the first monoclonal antibody and the anti-mullerian hormone are:
Glu Thr Tyr Gln Ala Asn Asn Cys Gln Gly Val Cys Gly Trp Pro Gln Ser Asp Arg Asn。
4. the kit according to claim 1,
the binding site of the second monoclonal antibody and the anti-mullerian hormone is located in the sequence shown in SEQ ID NO. 17.
5. The kit according to claim 4,
the binding sites of the second monoclonal antibody and the anti-mullerian hormone are:
Leu Gln Gly Leu Arg Val Glu Trp Arg Gly Arg Asp Pro Arg Gly Pro Gly Arg Ala Gln。
6. the kit according to claim 1,
the kit comprises a capture antibody and a detection antibody;
wherein the capture antibody is crosslinked with the magnetic bead, and the detection antibody is crosslinked with the luminescent label;
when either of the first monoclonal antibody and the second monoclonal antibody is a capture antibody, the other is a detection antibody.
7. A monoclonal antibody, comprising:
a first monoclonal antibody produced by a hybridoma cell strain with the preservation number of CGMCC NO. 23868; or
A second monoclonal antibody produced by a hybridoma cell strain with the preservation number of CGMCC NO. 23869.
8. The monoclonal antibody according to claim 7,
the monoclonal antibody is used for preparing an immunodetection tool for detecting anti-mullerian hormone.
9. A hybridoma cell line characterized in that,
the preservation number of the hybridoma cell strain is CGMCC NO. 23868; or
The preservation number of the hybridoma cell strain is CGMCC NO. 23869.
10. The monoclonal antibody according to claim 9,
the hybridoma cell strain is obtained by screening polypeptide;
the polypeptide is selected from the following sequences: SEQ ID NO.1 to SEQ ID NO. 21.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210324182.2A CN114609394B (en) | 2022-03-30 | 2022-03-30 | Anti-mullerian hormone detection kit, monoclonal antibody and hybridoma cell strain |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210324182.2A CN114609394B (en) | 2022-03-30 | 2022-03-30 | Anti-mullerian hormone detection kit, monoclonal antibody and hybridoma cell strain |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114609394A true CN114609394A (en) | 2022-06-10 |
| CN114609394B CN114609394B (en) | 2024-08-30 |
Family
ID=81867233
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210324182.2A Active CN114609394B (en) | 2022-03-30 | 2022-03-30 | Anti-mullerian hormone detection kit, monoclonal antibody and hybridoma cell strain |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114609394B (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013155883A1 (en) * | 2012-04-20 | 2013-10-24 | 中国农业科学院油料作物研究所 | Hybridoma cell line 3g1 and monoclonal antibody produced thereby against aflatoxin b1 |
| CN111273042A (en) * | 2020-01-21 | 2020-06-12 | 迈克生物股份有限公司 | Anti mullerian tube hormone detect reagent box |
| CN114075552A (en) * | 2022-01-19 | 2022-02-22 | 迈杰转化医学研究(苏州)有限公司 | Hybridoma cell strain secreting anti-FGL 1 monoclonal antibody and application thereof |
-
2022
- 2022-03-30 CN CN202210324182.2A patent/CN114609394B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013155883A1 (en) * | 2012-04-20 | 2013-10-24 | 中国农业科学院油料作物研究所 | Hybridoma cell line 3g1 and monoclonal antibody produced thereby against aflatoxin b1 |
| CN111273042A (en) * | 2020-01-21 | 2020-06-12 | 迈克生物股份有限公司 | Anti mullerian tube hormone detect reagent box |
| CN114075552A (en) * | 2022-01-19 | 2022-02-22 | 迈杰转化医学研究(苏州)有限公司 | Hybridoma cell strain secreting anti-FGL 1 monoclonal antibody and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| EZZAT, NOURIZADEH;SEYED JALAL, ZARGAR;MOHAMMAD HOSSEIN, ALIMOHAMMADIAN;SOHEILA, AJDARY;MAHDI, MAHDAVI: "Development of monoclonal antibodies against axenic amastigotes of Leishmania infantum strain in Iran: implication for diagnosis of Kala-azar.", IRANIAN JOURNAL OF BASIC MEDICAL SCIENCES, vol. 21, no. 4, 31 December 2018 (2018-12-31) * |
| 苏晓菱;李丽蓥;李苗苗;何洁;王瑞雪;王勇;罗树红;: "抗苗勒管激素的单克隆抗体制备与鉴定", 分子诊断与治疗杂志, no. 02, 18 February 2020 (2020-02-18) * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114609394B (en) | 2024-08-30 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| JP5743550B2 (en) | Antibody to N-terminal region of hemoglobin β chain | |
| CN111217907B (en) | anti-AMH monoclonal antibody and application thereof | |
| CN109912713B (en) | Preparation method of troponin I antibody for preparing immunodiagnostic reagent and obtained bacterial strain | |
| JP2729159B2 (en) | Monoclonal antibody of human glicentin, hybridoma producing the antibody, and method for quantifying human glicentin using the same | |
| CN113372447A (en) | anti-PIVKA-II monoclonal antibody and application thereof | |
| CA2785678C (en) | Human insulin assay and assay reagent | |
| CN119192368B (en) | Monoclonal antibody or antigen-binding fragment thereof for detecting folic acid and its binding protein complex, preparation method and application thereof | |
| CN114369160B (en) | Binding anti Lees' tube hormone AMH N High affinity rabbit monoclonal antibodies to dimeric proteins | |
| CN109705221B (en) | C peptide immunogen, monoclonal antibody pair thereof and application of antibody pair in C peptide magnetic particle chemiluminescence immunoassay reagent | |
| CN118344485A (en) | Anti-folic acid and monoclonal antibody of binding protein complex thereof or antigen binding fragment thereof, preparation method and application thereof | |
| CN114609394B (en) | Anti-mullerian hormone detection kit, monoclonal antibody and hybridoma cell strain | |
| CN113912719B (en) | Monoclonal antibody for detecting mouse interleukin 6 and preparation method and application thereof | |
| CN116813742A (en) | Human GDF-15 epitope peptide, antigen, antibody, kit and application | |
| WO2023193376A1 (en) | Pinellia ternata lectin enzyme-linked immunosorbent assay (elisa) kit and application | |
| JP4663831B2 (en) | Monoclonal antibodies, cell lines, and methods for measuring N1, N12-diacetylspermine | |
| CN114773466A (en) | Anti-human interleukin 23, kit comprising same and detection method thereof | |
| CN118005797B (en) | Anti-human alkaline phosphatase monoclonal antibody and application thereof in preparation of immune diagnostic reagent blocking agent | |
| CN114891104B (en) | Monoclonal antibody for identifying AMH and application thereof | |
| US20250035618A1 (en) | Detection method and detection reagent | |
| CN114686444B (en) | Hybridoma cell, antithrombotic regulatory protein monoclonal antibody, preparation method and application thereof | |
| CN118459581B (en) | Monoclonal antibody of anti-human oxidized low density lipoprotein and application thereof | |
| CN114686443B (en) | Hybridoma cell, antithrombotic regulatory protein monoclonal antibody, preparation method and application thereof | |
| CN119161475A (en) | A monoclonal antibody pair for detecting IGFBP2 protein or IGFBP2 polypeptide and its application | |
| CN117362432A (en) | C peptide recombinant rabbit monoclonal antibody, preparation method and application thereof | |
| CN115838692A (en) | Anti-human mutant erythropoietin mouse monoclonal antibody, application and human recombinant erythropoietin stimulant detection method |
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 |