CN112851824A - 一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用 - Google Patents
一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用 Download PDFInfo
- Publication number
- CN112851824A CN112851824A CN202110180894.7A CN202110180894A CN112851824A CN 112851824 A CN112851824 A CN 112851824A CN 202110180894 A CN202110180894 A CN 202110180894A CN 112851824 A CN112851824 A CN 112851824A
- Authority
- CN
- China
- Prior art keywords
- rbd
- protein
- fusion protein
- region
- val
- 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
- 102000037865 fusion proteins Human genes 0.000 title claims abstract description 21
- 108020001507 fusion proteins Proteins 0.000 title claims abstract description 21
- 241000711573 Coronaviridae Species 0.000 title claims abstract description 17
- 229940031626 subunit vaccine Drugs 0.000 title claims abstract description 9
- 238000002360 preparation method Methods 0.000 title claims description 7
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 25
- 102100031673 Corneodesmosin Human genes 0.000 claims abstract description 16
- 101710139375 Corneodesmosin Proteins 0.000 claims abstract description 16
- 241001678559 COVID-19 virus Species 0.000 claims abstract description 14
- 230000004927 fusion Effects 0.000 claims abstract description 10
- 108010076504 Protein Sorting Signals Proteins 0.000 claims description 10
- 108091026890 Coding region Proteins 0.000 claims description 7
- 238000011144 upstream manufacturing Methods 0.000 claims description 5
- 241001529936 Murinae Species 0.000 claims description 2
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 abstract description 22
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 abstract description 22
- 229940126583 recombinant protein vaccine Drugs 0.000 abstract description 19
- 102000004169 proteins and genes Human genes 0.000 abstract description 14
- 229960005486 vaccine Drugs 0.000 abstract description 14
- 230000003472 neutralizing effect Effects 0.000 abstract description 13
- 238000000034 method Methods 0.000 abstract description 8
- 230000008569 process Effects 0.000 abstract description 7
- 206010035664 Pneumonia Diseases 0.000 abstract description 4
- 108060003951 Immunoglobulin Proteins 0.000 abstract description 3
- 239000000539 dimer Substances 0.000 abstract description 3
- 102000018358 immunoglobulin Human genes 0.000 abstract description 3
- 230000007969 cellular immunity Effects 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 16
- 241000699666 Mus <mouse, genus> Species 0.000 description 11
- 241000699670 Mus sp. Species 0.000 description 11
- 150000001413 amino acids Chemical group 0.000 description 9
- 230000003053 immunization Effects 0.000 description 8
- 238000002649 immunization Methods 0.000 description 8
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 7
- 230000005847 immunogenicity Effects 0.000 description 6
- 239000000047 product Substances 0.000 description 6
- HKZAAJSTFUZYTO-LURJTMIESA-N (2s)-2-[[2-[[2-[[2-[(2-aminoacetyl)amino]acetyl]amino]acetyl]amino]acetyl]amino]-3-hydroxypropanoic acid Chemical compound NCC(=O)NCC(=O)NCC(=O)NCC(=O)N[C@@H](CO)C(O)=O HKZAAJSTFUZYTO-LURJTMIESA-N 0.000 description 5
- 108020004414 DNA Proteins 0.000 description 5
- 108090000174 Interleukin-10 Proteins 0.000 description 5
- 102000003814 Interleukin-10 Human genes 0.000 description 5
- 108090000978 Interleukin-4 Proteins 0.000 description 5
- 238000001514 detection method Methods 0.000 description 5
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 5
- 230000004048 modification Effects 0.000 description 5
- 238000012986 modification Methods 0.000 description 5
- 108020003175 receptors Proteins 0.000 description 5
- 102000005962 receptors Human genes 0.000 description 5
- 210000002966 serum Anatomy 0.000 description 5
- 102100035765 Angiotensin-converting enzyme 2 Human genes 0.000 description 4
- 108090000975 Angiotensin-converting enzyme 2 Proteins 0.000 description 4
- 241000283707 Capra Species 0.000 description 4
- 210000000170 cell membrane Anatomy 0.000 description 4
- 230000013595 glycosylation Effects 0.000 description 4
- 238000006206 glycosylation reaction Methods 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 239000002808 molecular sieve Substances 0.000 description 4
- URGAHOPLAPQHLN-UHFFFAOYSA-N sodium aluminosilicate Chemical compound [Na+].[Al+3].[O-][Si]([O-])=O.[O-][Si]([O-])=O URGAHOPLAPQHLN-UHFFFAOYSA-N 0.000 description 4
- 108090000695 Cytokines Proteins 0.000 description 3
- 102000004127 Cytokines Human genes 0.000 description 3
- 238000002965 ELISA Methods 0.000 description 3
- 241000124008 Mammalia Species 0.000 description 3
- VPEVBAUSTBWQHN-NHCYSSNCSA-N Pro-Glu-Val Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O VPEVBAUSTBWQHN-NHCYSSNCSA-N 0.000 description 3
- 102000044437 S1 domains Human genes 0.000 description 3
- 108700036684 S1 domains Proteins 0.000 description 3
- 108091005634 SARS-CoV-2 receptor-binding domains Proteins 0.000 description 3
- 241000700605 Viruses Species 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 238000012809 post-inoculation Methods 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 210000004989 spleen cell Anatomy 0.000 description 3
- 210000004988 splenocyte Anatomy 0.000 description 3
- 230000008685 targeting Effects 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
- XMIAMUXIMWREBJ-HERUPUMHSA-N Ala-Trp-Asn Chemical compound C[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)N[C@@H](CC(=O)N)C(=O)O)N XMIAMUXIMWREBJ-HERUPUMHSA-N 0.000 description 2
- ALTQTAKGRFLRLR-GUBZILKMSA-N Cys-Val-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CS)N ALTQTAKGRFLRLR-GUBZILKMSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 241000588724 Escherichia coli Species 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 102000018071 Immunoglobulin Fc Fragments Human genes 0.000 description 2
- 108010091135 Immunoglobulin Fc Fragments Proteins 0.000 description 2
- 102100037850 Interferon gamma Human genes 0.000 description 2
- 108010074328 Interferon-gamma Proteins 0.000 description 2
- HNDMFDBQXYZSRM-IHRRRGAJSA-N Ser-Val-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O HNDMFDBQXYZSRM-IHRRRGAJSA-N 0.000 description 2
- 108010008038 Synthetic Vaccines Proteins 0.000 description 2
- 230000024932 T cell mediated immunity Effects 0.000 description 2
- SKHPKKYKDYULDH-HJGDQZAQSA-N Thr-Asn-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O SKHPKKYKDYULDH-HJGDQZAQSA-N 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
- 108010003533 Viral Envelope Proteins Proteins 0.000 description 2
- 239000002671 adjuvant Substances 0.000 description 2
- 108010047857 aspartylglycine Proteins 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000005520 cutting process Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000000684 flow cytometry Methods 0.000 description 2
- 108010010147 glycylglutamine Proteins 0.000 description 2
- 238000011534 incubation Methods 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 210000003141 lower extremity Anatomy 0.000 description 2
- 108010064235 lysylglycine Proteins 0.000 description 2
- 108010017391 lysylvaline Proteins 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 210000004779 membrane envelope Anatomy 0.000 description 2
- 230000004481 post-translational protein modification Effects 0.000 description 2
- 210000001236 prokaryotic cell Anatomy 0.000 description 2
- 108010077112 prolyl-proline Proteins 0.000 description 2
- 108010031719 prolyl-serine Proteins 0.000 description 2
- 229940023143 protein vaccine Drugs 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000012264 purified product Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 108010048818 seryl-histidine Proteins 0.000 description 2
- 238000003998 size exclusion chromatography high performance liquid chromatography Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 108010061238 threonyl-glycine Proteins 0.000 description 2
- 230000014621 translational initiation Effects 0.000 description 2
- 239000013638 trimer Substances 0.000 description 2
- 108010051110 tyrosyl-lysine Proteins 0.000 description 2
- CWFMWBHMIMNZLN-NAKRPEOUSA-N (2s)-1-[(2s)-2-[[(2s,3s)-2-amino-3-methylpentanoyl]amino]propanoyl]pyrrolidine-2-carboxylic acid Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C)C(=O)N1CCC[C@H]1C(O)=O CWFMWBHMIMNZLN-NAKRPEOUSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- WRDANSJTFOHBPI-FXQIFTODSA-N Ala-Arg-Cys Chemical compound C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CS)C(=O)O)N WRDANSJTFOHBPI-FXQIFTODSA-N 0.000 description 1
- BUDNAJYVCUHLSV-ZLUOBGJFSA-N Ala-Asp-Ser Chemical compound C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O BUDNAJYVCUHLSV-ZLUOBGJFSA-N 0.000 description 1
- BTYTYHBSJKQBQA-GCJQMDKQSA-N Ala-Asp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](C)N)O BTYTYHBSJKQBQA-GCJQMDKQSA-N 0.000 description 1
- KUDREHRZRIVKHS-UWJYBYFXSA-N Ala-Asp-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KUDREHRZRIVKHS-UWJYBYFXSA-N 0.000 description 1
- SFNFGFDRYJKZKN-XQXXSGGOSA-N Ala-Gln-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](C)N)O SFNFGFDRYJKZKN-XQXXSGGOSA-N 0.000 description 1
- DPNZTBKGAUAZQU-DLOVCJGASA-N Ala-Leu-His Chemical compound C[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)N DPNZTBKGAUAZQU-DLOVCJGASA-N 0.000 description 1
- OINVDEKBKBCPLX-JXUBOQSCSA-N Ala-Lys-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OINVDEKBKBCPLX-JXUBOQSCSA-N 0.000 description 1
- WQKAQKZRDIZYNV-VZFHVOOUSA-N Ala-Ser-Thr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WQKAQKZRDIZYNV-VZFHVOOUSA-N 0.000 description 1
- IETUUAHKCHOQHP-KZVJFYERSA-N Ala-Thr-Val Chemical compound CC(C)[C@H](NC(=O)[C@@H](NC(=O)[C@H](C)N)[C@@H](C)O)C(O)=O IETUUAHKCHOQHP-KZVJFYERSA-N 0.000 description 1
- REWSWYIDQIELBE-FXQIFTODSA-N Ala-Val-Ser Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O REWSWYIDQIELBE-FXQIFTODSA-N 0.000 description 1
- IYMAXBFPHPZYIK-BQBZGAKWSA-N Arg-Gly-Asp Chemical compound NC(N)=NCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(O)=O IYMAXBFPHPZYIK-BQBZGAKWSA-N 0.000 description 1
- FSNVAJOPUDVQAR-AVGNSLFASA-N Arg-Lys-Arg Chemical compound NC(=N)NCCC[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O FSNVAJOPUDVQAR-AVGNSLFASA-N 0.000 description 1
- CZUHPNLXLWMYMG-UBHSHLNASA-N Arg-Phe-Ala Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C)C(O)=O)CC1=CC=CC=C1 CZUHPNLXLWMYMG-UBHSHLNASA-N 0.000 description 1
- ZJBUILVYSXQNSW-YTWAJWBKSA-N Arg-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N)O ZJBUILVYSXQNSW-YTWAJWBKSA-N 0.000 description 1
- PSUXEQYPYZLNER-QXEWZRGKSA-N Arg-Val-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O PSUXEQYPYZLNER-QXEWZRGKSA-N 0.000 description 1
- BVLIJXXSXBUGEC-SRVKXCTJSA-N Asn-Asn-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BVLIJXXSXBUGEC-SRVKXCTJSA-N 0.000 description 1
- SRUUBQBAVNQZGJ-LAEOZQHASA-N Asn-Gln-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC(=O)N)N SRUUBQBAVNQZGJ-LAEOZQHASA-N 0.000 description 1
- HYQYLOSCICEYTR-YUMQZZPRSA-N Asn-Gly-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](CC(C)C)C(O)=O HYQYLOSCICEYTR-YUMQZZPRSA-N 0.000 description 1
- OLVIPTLKNSAYRJ-YUMQZZPRSA-N Asn-Gly-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)CNC(=O)[C@H](CC(=O)N)N OLVIPTLKNSAYRJ-YUMQZZPRSA-N 0.000 description 1
- ZTRJUKDEALVRMW-SRVKXCTJSA-N Asn-His-His Chemical compound C1=C(NC=N1)C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CC(=O)N)N ZTRJUKDEALVRMW-SRVKXCTJSA-N 0.000 description 1
- WQLJRNRLHWJIRW-KKUMJFAQSA-N Asn-His-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC2=CN=CN2)NC(=O)[C@H](CC(=O)N)N)O WQLJRNRLHWJIRW-KKUMJFAQSA-N 0.000 description 1
- RCFGLXMZDYNRSC-CIUDSAMLSA-N Asn-Lys-Ala Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](C)C(O)=O RCFGLXMZDYNRSC-CIUDSAMLSA-N 0.000 description 1
- PIABYSIYPGLLDQ-XVSYOHENSA-N Asn-Thr-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O PIABYSIYPGLLDQ-XVSYOHENSA-N 0.000 description 1
- RTFXPCYMDYBZNQ-SRVKXCTJSA-N Asn-Tyr-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O RTFXPCYMDYBZNQ-SRVKXCTJSA-N 0.000 description 1
- QNNBHTFDFFFHGC-KKUMJFAQSA-N Asn-Tyr-Lys Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O QNNBHTFDFFFHGC-KKUMJFAQSA-N 0.000 description 1
- LTDGPJKGJDIBQD-LAEOZQHASA-N Asn-Val-Gln Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(N)=O)C(O)=O LTDGPJKGJDIBQD-LAEOZQHASA-N 0.000 description 1
- KBQOUDLMWYWXNP-YDHLFZDLSA-N Asn-Val-Phe Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CC(=O)N)N KBQOUDLMWYWXNP-YDHLFZDLSA-N 0.000 description 1
- HBUJSDCLZCXXCW-YDHLFZDLSA-N Asn-Val-Tyr Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HBUJSDCLZCXXCW-YDHLFZDLSA-N 0.000 description 1
- PXLNPFOJZQMXAT-BYULHYEWSA-N Asp-Asp-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CC(O)=O PXLNPFOJZQMXAT-BYULHYEWSA-N 0.000 description 1
- KLYPOCBLKMPBIQ-GHCJXIJMSA-N Asp-Ile-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)O)NC(=O)[C@H](CC(=O)O)N KLYPOCBLKMPBIQ-GHCJXIJMSA-N 0.000 description 1
- SPWXXPFDTMYTRI-IUKAMOBKSA-N Asp-Ile-Thr Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SPWXXPFDTMYTRI-IUKAMOBKSA-N 0.000 description 1
- CUQDCPXNZPDYFQ-ZLUOBGJFSA-N Asp-Ser-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O CUQDCPXNZPDYFQ-ZLUOBGJFSA-N 0.000 description 1
- VNXQRBXEQXLERQ-CIUDSAMLSA-N Asp-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)O)N VNXQRBXEQXLERQ-CIUDSAMLSA-N 0.000 description 1
- IWLZBRTUIVXZJD-OLHMAJIHSA-N Asp-Thr-Asp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O IWLZBRTUIVXZJD-OLHMAJIHSA-N 0.000 description 1
- JSNWZMFSLIWAHS-HJGDQZAQSA-N Asp-Thr-Leu Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H](CC(=O)O)N)O JSNWZMFSLIWAHS-HJGDQZAQSA-N 0.000 description 1
- YUELDQUPTAYEGM-XIRDDKMYSA-N Asp-Trp-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CC(=O)O)N YUELDQUPTAYEGM-XIRDDKMYSA-N 0.000 description 1
- GIKOVDMXBAFXDF-NHCYSSNCSA-N Asp-Val-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O GIKOVDMXBAFXDF-NHCYSSNCSA-N 0.000 description 1
- QOJJMJKTMKNFEF-ZKWXMUAHSA-N Asp-Val-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC(O)=O QOJJMJKTMKNFEF-ZKWXMUAHSA-N 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 208000025721 COVID-19 Diseases 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- OIMUAKUQOUEPCZ-WHFBIAKZSA-N Cys-Asn-Gly Chemical compound SC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O OIMUAKUQOUEPCZ-WHFBIAKZSA-N 0.000 description 1
- YMBAVNPKBWHDAW-CIUDSAMLSA-N Cys-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CS)N YMBAVNPKBWHDAW-CIUDSAMLSA-N 0.000 description 1
- SKSJPIBFNFPTJB-NKWVEPMBSA-N Cys-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CS)N)C(=O)O SKSJPIBFNFPTJB-NKWVEPMBSA-N 0.000 description 1
- OHLLDUNVMPPUMD-DCAQKATOSA-N Cys-Leu-Val Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CS)N OHLLDUNVMPPUMD-DCAQKATOSA-N 0.000 description 1
- PEZINYWZBQNTIX-NAKRPEOUSA-N Cys-Met-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)N PEZINYWZBQNTIX-NAKRPEOUSA-N 0.000 description 1
- SMEYEQDCCBHTEF-FXQIFTODSA-N Cys-Pro-Ala Chemical compound [H]N[C@@H](CS)C(=O)N1CCC[C@H]1C(=O)N[C@@H](C)C(O)=O SMEYEQDCCBHTEF-FXQIFTODSA-N 0.000 description 1
- CNAMJJOZGXPDHW-IHRRRGAJSA-N Cys-Pro-Phe Chemical compound N[C@@H](CS)C(=O)N1CCC[C@H]1C(=O)N[C@@H](Cc1ccccc1)C(O)=O CNAMJJOZGXPDHW-IHRRRGAJSA-N 0.000 description 1
- KSMSFCBQBQPFAD-GUBZILKMSA-N Cys-Pro-Pro Chemical compound SC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 KSMSFCBQBQPFAD-GUBZILKMSA-N 0.000 description 1
- NDNZRWUDUMTITL-FXQIFTODSA-N Cys-Ser-Val Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NDNZRWUDUMTITL-FXQIFTODSA-N 0.000 description 1
- KFYPRIGJTICABD-XGEHTFHBSA-N Cys-Thr-Val Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)O)NC(=O)[C@H](CS)N)O KFYPRIGJTICABD-XGEHTFHBSA-N 0.000 description 1
- JRZMCSIUYGSJKP-ZKWXMUAHSA-N Cys-Val-Asn Chemical compound SC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O JRZMCSIUYGSJKP-ZKWXMUAHSA-N 0.000 description 1
- NGOIQDYZMIKCOK-NAKRPEOUSA-N Cys-Val-Ile Chemical compound [H]N[C@@H](CS)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NGOIQDYZMIKCOK-NAKRPEOUSA-N 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 101710091045 Envelope protein Proteins 0.000 description 1
- 102000004961 Furin Human genes 0.000 description 1
- 108090001126 Furin Proteins 0.000 description 1
- HHWQMFIGMMOVFK-WDSKDSINSA-N Gln-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(N)=O HHWQMFIGMMOVFK-WDSKDSINSA-N 0.000 description 1
- NVEASDQHBRZPSU-BQBZGAKWSA-N Gln-Gln-Gly Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)NCC(O)=O NVEASDQHBRZPSU-BQBZGAKWSA-N 0.000 description 1
- LVNILKSSFHCSJZ-IHRRRGAJSA-N Gln-Gln-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCC(=O)N)N LVNILKSSFHCSJZ-IHRRRGAJSA-N 0.000 description 1
- KLKYKPXITJBSNI-CIUDSAMLSA-N Gln-Met-Ala Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](C)C(O)=O KLKYKPXITJBSNI-CIUDSAMLSA-N 0.000 description 1
- UESYBOXFJWJVSB-AVGNSLFASA-N Gln-Phe-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O UESYBOXFJWJVSB-AVGNSLFASA-N 0.000 description 1
- FNAJNWPDTIXYJN-CIUDSAMLSA-N Gln-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CCC(N)=O FNAJNWPDTIXYJN-CIUDSAMLSA-N 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
- HMIXCETWRYDVMO-GUBZILKMSA-N Gln-Pro-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(O)=O)C(O)=O HMIXCETWRYDVMO-GUBZILKMSA-N 0.000 description 1
- OREPWMPAUWIIAM-ZPFDUUQYSA-N Gln-Pro-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@@H]1CCCN1C(=O)[C@H](CCC(=O)N)N OREPWMPAUWIIAM-ZPFDUUQYSA-N 0.000 description 1
- GHAXJVNBAKGWEJ-AVGNSLFASA-N Gln-Ser-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O GHAXJVNBAKGWEJ-AVGNSLFASA-N 0.000 description 1
- CSMHMEATMDCQNY-DZKIICNBSA-N Gln-Val-Tyr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O CSMHMEATMDCQNY-DZKIICNBSA-N 0.000 description 1
- RUFHOVYUYSNDNY-ACZMJKKPSA-N Glu-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(O)=O RUFHOVYUYSNDNY-ACZMJKKPSA-N 0.000 description 1
- UTKUTMJSWKKHEM-WDSKDSINSA-N Glu-Ala-Gly Chemical compound OC(=O)CNC(=O)[C@H](C)NC(=O)[C@@H](N)CCC(O)=O UTKUTMJSWKKHEM-WDSKDSINSA-N 0.000 description 1
- ZJICFHQSPWFBKP-AVGNSLFASA-N Glu-Asn-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ZJICFHQSPWFBKP-AVGNSLFASA-N 0.000 description 1
- HUFCEIHAFNVSNR-IHRRRGAJSA-N Glu-Gln-Tyr Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 HUFCEIHAFNVSNR-IHRRRGAJSA-N 0.000 description 1
- HNVFSTLPVJWIDV-CIUDSAMLSA-N Glu-Glu-Gln Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(N)=O)C(O)=O HNVFSTLPVJWIDV-CIUDSAMLSA-N 0.000 description 1
- NJCALAAIGREHDR-WDCWCFNPSA-N Glu-Leu-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NJCALAAIGREHDR-WDCWCFNPSA-N 0.000 description 1
- AAJHGGDRKHYSDH-GUBZILKMSA-N Glu-Pro-Gln Chemical compound C1C[C@H](N(C1)C(=O)[C@H](CCC(=O)O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O AAJHGGDRKHYSDH-GUBZILKMSA-N 0.000 description 1
- ALMBZBOCGSVSAI-ACZMJKKPSA-N Glu-Ser-Asn Chemical compound C(CC(=O)O)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)N)C(=O)O)N ALMBZBOCGSVSAI-ACZMJKKPSA-N 0.000 description 1
- BPCLDCNZBUYGOD-BPUTZDHNSA-N Glu-Trp-Glu Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@H](CCC(O)=O)N)C(=O)N[C@@H](CCC(O)=O)C(O)=O)=CNC2=C1 BPCLDCNZBUYGOD-BPUTZDHNSA-N 0.000 description 1
- MLILEEIVMRUYBX-NHCYSSNCSA-N Glu-Val-Arg Chemical compound OC(=O)CC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(O)=O MLILEEIVMRUYBX-NHCYSSNCSA-N 0.000 description 1
- ZALGPUWUVHOGAE-GVXVVHGQSA-N Glu-Val-His Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](CCC(=O)O)N ZALGPUWUVHOGAE-GVXVVHGQSA-N 0.000 description 1
- WGYHAAXZWPEBDQ-IFFSRLJSSA-N Glu-Val-Thr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WGYHAAXZWPEBDQ-IFFSRLJSSA-N 0.000 description 1
- QXPRJQPCFXMCIY-NKWVEPMBSA-N Gly-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)CN QXPRJQPCFXMCIY-NKWVEPMBSA-N 0.000 description 1
- CLODWIOAKCSBAN-BQBZGAKWSA-N Gly-Arg-Asp Chemical compound NC(N)=NCCC[C@H](NC(=O)CN)C(=O)N[C@@H](CC(O)=O)C(O)=O CLODWIOAKCSBAN-BQBZGAKWSA-N 0.000 description 1
- VXKCPBPQEKKERH-IUCAKERBSA-N Gly-Arg-Pro Chemical compound NC(N)=NCCC[C@H](NC(=O)CN)C(=O)N1CCC[C@H]1C(O)=O VXKCPBPQEKKERH-IUCAKERBSA-N 0.000 description 1
- KKBWDNZXYLGJEY-UHFFFAOYSA-N Gly-Arg-Pro Natural products NCC(=O)NC(CCNC(=N)N)C(=O)N1CCCC1C(=O)O KKBWDNZXYLGJEY-UHFFFAOYSA-N 0.000 description 1
- LGQZOQRDEUIZJY-YUMQZZPRSA-N Gly-Cys-Lys Chemical compound NCCCC[C@H](NC(=O)[C@H](CS)NC(=O)CN)C(O)=O LGQZOQRDEUIZJY-YUMQZZPRSA-N 0.000 description 1
- NPSWCZIRBAYNSB-JHEQGTHGSA-N Gly-Gln-Thr Chemical compound [H]NCC(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NPSWCZIRBAYNSB-JHEQGTHGSA-N 0.000 description 1
- JSNNHGHYGYMVCK-XVKPBYJWSA-N Gly-Glu-Val Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O JSNNHGHYGYMVCK-XVKPBYJWSA-N 0.000 description 1
- BUEFQXUHTUZXHR-LURJTMIESA-N Gly-Gly-Pro zwitterion Chemical compound NCC(=O)NCC(=O)N1CCC[C@H]1C(O)=O BUEFQXUHTUZXHR-LURJTMIESA-N 0.000 description 1
- LIXWIUAORXJNBH-QWRGUYRKSA-N Gly-Leu-His Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)CN LIXWIUAORXJNBH-QWRGUYRKSA-N 0.000 description 1
- GMTXWRIDLGTVFC-IUCAKERBSA-N Gly-Lys-Glu Chemical compound [H]NCC(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O GMTXWRIDLGTVFC-IUCAKERBSA-N 0.000 description 1
- PTIIBFKSLCYQBO-NHCYSSNCSA-N Gly-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)CN PTIIBFKSLCYQBO-NHCYSSNCSA-N 0.000 description 1
- YYXJFBMCOUSYSF-RYUDHWBXSA-N Gly-Phe-Gln Chemical compound [H]NCC(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(N)=O)C(O)=O YYXJFBMCOUSYSF-RYUDHWBXSA-N 0.000 description 1
- JSLVAHYTAJJEQH-QWRGUYRKSA-N Gly-Ser-Phe Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 JSLVAHYTAJJEQH-QWRGUYRKSA-N 0.000 description 1
- FKYQEVBRZSFAMJ-QWRGUYRKSA-N Gly-Ser-Tyr Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 FKYQEVBRZSFAMJ-QWRGUYRKSA-N 0.000 description 1
- SYOJVRNQCXYEOV-XVKPBYJWSA-N Gly-Val-Glu Chemical compound [H]NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O SYOJVRNQCXYEOV-XVKPBYJWSA-N 0.000 description 1
- RYAOJUMWLWUGNW-QMMMGPOBSA-N Gly-Val-Gly Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)NCC(O)=O RYAOJUMWLWUGNW-QMMMGPOBSA-N 0.000 description 1
- BAYQNCWLXIDLHX-ONGXEEELSA-N Gly-Val-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN BAYQNCWLXIDLHX-ONGXEEELSA-N 0.000 description 1
- SBVMXEZQJVUARN-XPUUQOCRSA-N Gly-Val-Ser Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O SBVMXEZQJVUARN-XPUUQOCRSA-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
- SDTPKSOWFXBACN-GUBZILKMSA-N His-Glu-Asp Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O SDTPKSOWFXBACN-GUBZILKMSA-N 0.000 description 1
- GIRSNERMXCMDBO-GARJFASQSA-N His-Ser-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CO)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O GIRSNERMXCMDBO-GARJFASQSA-N 0.000 description 1
- 102100026120 IgG receptor FcRn large subunit p51 Human genes 0.000 description 1
- 101710177940 IgG receptor FcRn large subunit p51 Proteins 0.000 description 1
- MKWSZEHGHSLNPF-NAKRPEOUSA-N Ile-Ala-Val Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](C)C(=O)N[C@@H](C(C)C)C(=O)O)N MKWSZEHGHSLNPF-NAKRPEOUSA-N 0.000 description 1
- DFJJAVZIHDFOGQ-MNXVOIDGSA-N Ile-Glu-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N DFJJAVZIHDFOGQ-MNXVOIDGSA-N 0.000 description 1
- QQFSKBMCAKWHLG-UHFFFAOYSA-N Ile-Phe-Pro-Pro Chemical compound C1CCC(C(=O)N2C(CCC2)C(O)=O)N1C(=O)C(NC(=O)C(N)C(C)CC)CC1=CC=CC=C1 QQFSKBMCAKWHLG-UHFFFAOYSA-N 0.000 description 1
- FQYQMFCIJNWDQZ-CYDGBPFRSA-N Ile-Pro-Pro Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 FQYQMFCIJNWDQZ-CYDGBPFRSA-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
- 102220622573 Inositol-tetrakisphosphate 1-kinase_N297L_mutation Human genes 0.000 description 1
- 108010065920 Insulin Lispro Proteins 0.000 description 1
- PMGDADKJMCOXHX-UHFFFAOYSA-N L-Arginyl-L-glutamin-acetat Natural products NC(=N)NCCCC(N)C(=O)NC(CCC(N)=O)C(O)=O PMGDADKJMCOXHX-UHFFFAOYSA-N 0.000 description 1
- 241000880493 Leptailurus serval Species 0.000 description 1
- VCSBGUACOYUIGD-CIUDSAMLSA-N Leu-Asn-Asp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O VCSBGUACOYUIGD-CIUDSAMLSA-N 0.000 description 1
- YORLGJINWYYIMX-KKUMJFAQSA-N Leu-Cys-Phe Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O YORLGJINWYYIMX-KKUMJFAQSA-N 0.000 description 1
- BKTXKJMNTSMJDQ-AVGNSLFASA-N Leu-His-Gln Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N BKTXKJMNTSMJDQ-AVGNSLFASA-N 0.000 description 1
- RTIRBWJPYJYTLO-MELADBBJSA-N Leu-Lys-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N1CCC[C@@H]1C(=O)O)N RTIRBWJPYJYTLO-MELADBBJSA-N 0.000 description 1
- GCXGCIYIHXSKAY-ULQDDVLXSA-N Leu-Phe-Arg Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O GCXGCIYIHXSKAY-ULQDDVLXSA-N 0.000 description 1
- UHNQRAFSEBGZFZ-YESZJQIVSA-N Leu-Phe-Pro Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N2CCC[C@@H]2C(=O)O)N UHNQRAFSEBGZFZ-YESZJQIVSA-N 0.000 description 1
- YUTNOGOMBNYPFH-XUXIUFHCSA-N Leu-Pro-Ile Chemical compound [H]N[C@@H](CC(C)C)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(O)=O YUTNOGOMBNYPFH-XUXIUFHCSA-N 0.000 description 1
- XXXXOVFBXRERQL-ULQDDVLXSA-N Leu-Pro-Phe Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 XXXXOVFBXRERQL-ULQDDVLXSA-N 0.000 description 1
- DPURXCQCHSQPAN-AVGNSLFASA-N Leu-Pro-Pro Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 DPURXCQCHSQPAN-AVGNSLFASA-N 0.000 description 1
- XOWMDXHFSBCAKQ-SRVKXCTJSA-N Leu-Ser-Leu Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CC(C)C XOWMDXHFSBCAKQ-SRVKXCTJSA-N 0.000 description 1
- SBANPBVRHYIMRR-UHFFFAOYSA-N Leu-Ser-Pro Natural products CC(C)CC(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O SBANPBVRHYIMRR-UHFFFAOYSA-N 0.000 description 1
- KLSUAWUZBMAZCL-RHYQMDGZSA-N Leu-Thr-Pro Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(O)=O KLSUAWUZBMAZCL-RHYQMDGZSA-N 0.000 description 1
- AIQWYVFNBNNOLU-RHYQMDGZSA-N Leu-Thr-Val Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(O)=O AIQWYVFNBNNOLU-RHYQMDGZSA-N 0.000 description 1
- WUHBLPVELFTPQK-KKUMJFAQSA-N Leu-Tyr-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(O)=O WUHBLPVELFTPQK-KKUMJFAQSA-N 0.000 description 1
- WSXTWLJHTLRFLW-SRVKXCTJSA-N Lys-Ala-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CCCCN)C(O)=O WSXTWLJHTLRFLW-SRVKXCTJSA-N 0.000 description 1
- IXHKPDJKKCUKHS-GARJFASQSA-N Lys-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCCCN)N IXHKPDJKKCUKHS-GARJFASQSA-N 0.000 description 1
- QUCDKEKDPYISNX-HJGDQZAQSA-N Lys-Asn-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O QUCDKEKDPYISNX-HJGDQZAQSA-N 0.000 description 1
- LMVOVCYVZBBWQB-SRVKXCTJSA-N Lys-Asp-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CCCCN LMVOVCYVZBBWQB-SRVKXCTJSA-N 0.000 description 1
- DZQYZKPINJLLEN-KKUMJFAQSA-N Lys-Cys-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CCCCN)N)O DZQYZKPINJLLEN-KKUMJFAQSA-N 0.000 description 1
- OIQSIMFSVLLWBX-VOAKCMCISA-N Lys-Leu-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O OIQSIMFSVLLWBX-VOAKCMCISA-N 0.000 description 1
- ATNKHRAIZCMCCN-BZSNNMDCSA-N Lys-Lys-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CCCCN)N ATNKHRAIZCMCCN-BZSNNMDCSA-N 0.000 description 1
- YDDDRTIPNTWGIG-SRVKXCTJSA-N Lys-Lys-Ser Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O YDDDRTIPNTWGIG-SRVKXCTJSA-N 0.000 description 1
- ODTZHNZPINULEU-KKUMJFAQSA-N Lys-Phe-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N ODTZHNZPINULEU-KKUMJFAQSA-N 0.000 description 1
- LUTDBHBIHHREDC-IHRRRGAJSA-N Lys-Pro-Lys Chemical compound NCCCC[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(O)=O LUTDBHBIHHREDC-IHRRRGAJSA-N 0.000 description 1
- GHKXHCMRAUYLBS-CIUDSAMLSA-N Lys-Ser-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O GHKXHCMRAUYLBS-CIUDSAMLSA-N 0.000 description 1
- YKBSXQFZWFXFIB-VOAKCMCISA-N Lys-Thr-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@H](O)C)C(=O)N[C@@H](CCCCN)C(O)=O YKBSXQFZWFXFIB-VOAKCMCISA-N 0.000 description 1
- RIPJMCFGQHGHNP-RHYQMDGZSA-N Lys-Val-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](C(C)C)NC(=O)[C@H](CCCCN)N)O RIPJMCFGQHGHNP-RHYQMDGZSA-N 0.000 description 1
- 108010052285 Membrane Proteins Proteins 0.000 description 1
- 102000018697 Membrane Proteins Human genes 0.000 description 1
- CFRRIZLGFGJEDB-SRVKXCTJSA-N Met-His-Gln Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(N)=O)C(O)=O CFRRIZLGFGJEDB-SRVKXCTJSA-N 0.000 description 1
- RKIIYGUHIQJCBW-SRVKXCTJSA-N Met-His-Glu Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(O)=O RKIIYGUHIQJCBW-SRVKXCTJSA-N 0.000 description 1
- FWAHLGXNBLWIKB-NAKRPEOUSA-N Met-Ile-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCSC FWAHLGXNBLWIKB-NAKRPEOUSA-N 0.000 description 1
- 208000025370 Middle East respiratory syndrome Diseases 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
- SITLTJHOQZFJGG-UHFFFAOYSA-N N-L-alpha-glutamyl-L-valine Natural products CC(C)C(C(O)=O)NC(=O)C(N)CCC(O)=O SITLTJHOQZFJGG-UHFFFAOYSA-N 0.000 description 1
- 241001292005 Nidovirales Species 0.000 description 1
- 108700001237 Nucleic Acid-Based Vaccines Proteins 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 108090001074 Nucleocapsid Proteins Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 102000005877 Peptide Initiation Factors Human genes 0.000 description 1
- 108010044843 Peptide Initiation Factors Proteins 0.000 description 1
- QCHNRQQVLJYDSI-DLOVCJGASA-N Phe-Asn-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 QCHNRQQVLJYDSI-DLOVCJGASA-N 0.000 description 1
- UUWCIPUVJJIEEP-SRVKXCTJSA-N Phe-Asn-Cys Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CS)C(=O)O)N UUWCIPUVJJIEEP-SRVKXCTJSA-N 0.000 description 1
- AWAYOWOUGVZXOB-BZSNNMDCSA-N Phe-Asn-Phe Chemical compound C([C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(O)=O)C1=CC=CC=C1 AWAYOWOUGVZXOB-BZSNNMDCSA-N 0.000 description 1
- CDNPIRSCAFMMBE-SRVKXCTJSA-N Phe-Asn-Ser Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O CDNPIRSCAFMMBE-SRVKXCTJSA-N 0.000 description 1
- HOYQLNNGMHXZDW-KKUMJFAQSA-N Phe-Glu-Arg Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O HOYQLNNGMHXZDW-KKUMJFAQSA-N 0.000 description 1
- CJAHQEZWDZNSJO-KKUMJFAQSA-N Phe-Lys-Cys Chemical compound NCCCC[C@@H](C(=O)N[C@@H](CS)C(O)=O)NC(=O)[C@@H](N)CC1=CC=CC=C1 CJAHQEZWDZNSJO-KKUMJFAQSA-N 0.000 description 1
- JDMKQHSHKJHAHR-UHFFFAOYSA-N Phe-Phe-Leu-Tyr Natural products C=1C=C(O)C=CC=1CC(C(O)=O)NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)CC1=CC=CC=C1 JDMKQHSHKJHAHR-UHFFFAOYSA-N 0.000 description 1
- JLLJTMHNXQTMCK-UBHSHLNASA-N Phe-Pro-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=CC=C1 JLLJTMHNXQTMCK-UBHSHLNASA-N 0.000 description 1
- QARPMYDMYVLFMW-KKUMJFAQSA-N Phe-Pro-Glu Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCC(O)=O)C(O)=O)C1=CC=CC=C1 QARPMYDMYVLFMW-KKUMJFAQSA-N 0.000 description 1
- MMJJFXWMCMJMQA-STQMWFEESA-N Phe-Pro-Gly Chemical compound C([C@H](N)C(=O)N1[C@@H](CCC1)C(=O)NCC(O)=O)C1=CC=CC=C1 MMJJFXWMCMJMQA-STQMWFEESA-N 0.000 description 1
- WWPAHTZOWURIMR-ULQDDVLXSA-N Phe-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)CC1=CC=CC=C1 WWPAHTZOWURIMR-ULQDDVLXSA-N 0.000 description 1
- IAOZOFPONWDXNT-IXOXFDKPSA-N Phe-Ser-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O IAOZOFPONWDXNT-IXOXFDKPSA-N 0.000 description 1
- BSKMOCNNLNDIMU-CDMKHQONSA-N Phe-Thr-Gly Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O BSKMOCNNLNDIMU-CDMKHQONSA-N 0.000 description 1
- SJRQWEDYTKYHHL-SLFFLAALSA-N Phe-Tyr-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC2=CC=C(C=C2)O)NC(=O)[C@H](CC3=CC=CC=C3)N)C(=O)O SJRQWEDYTKYHHL-SLFFLAALSA-N 0.000 description 1
- BQMFWUKNOCJDNV-HJWJTTGWSA-N Phe-Val-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BQMFWUKNOCJDNV-HJWJTTGWSA-N 0.000 description 1
- GAMLAXHLYGLQBJ-UFYCRDLUSA-N Phe-Val-Tyr Chemical compound N[C@H](C(=O)N[C@H](C(=O)N[C@H](C(=O)O)CC1=CC=C(C=C1)O)C(C)C)CC1=CC=CC=C1 GAMLAXHLYGLQBJ-UFYCRDLUSA-N 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- IHCXPSYCHXFXKT-DCAQKATOSA-N Pro-Arg-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(O)=O IHCXPSYCHXFXKT-DCAQKATOSA-N 0.000 description 1
- XWYXZPHPYKRYPA-GMOBBJLQSA-N Pro-Asn-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O XWYXZPHPYKRYPA-GMOBBJLQSA-N 0.000 description 1
- ILMLVTGTUJPQFP-FXQIFTODSA-N Pro-Asp-Asp Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(O)=O)C(O)=O ILMLVTGTUJPQFP-FXQIFTODSA-N 0.000 description 1
- GQLOZEMWEBDEAY-NAKRPEOUSA-N Pro-Cys-Ile Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CS)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GQLOZEMWEBDEAY-NAKRPEOUSA-N 0.000 description 1
- XZONQWUEBAFQPO-HJGDQZAQSA-N Pro-Gln-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O XZONQWUEBAFQPO-HJGDQZAQSA-N 0.000 description 1
- KWMUAKQOVYCQJQ-ZPFDUUQYSA-N Pro-Ile-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@@H]1CCCN1 KWMUAKQOVYCQJQ-ZPFDUUQYSA-N 0.000 description 1
- XQPHBAKJJJZOBX-SRVKXCTJSA-N Pro-Lys-Glu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(O)=O XQPHBAKJJJZOBX-SRVKXCTJSA-N 0.000 description 1
- ULWBBFKQBDNGOY-RWMBFGLXSA-N Pro-Lys-Pro Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CCCCN)C(=O)N2CCC[C@@H]2C(=O)O ULWBBFKQBDNGOY-RWMBFGLXSA-N 0.000 description 1
- MHHQQZIFLWFZGR-DCAQKATOSA-N Pro-Lys-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O MHHQQZIFLWFZGR-DCAQKATOSA-N 0.000 description 1
- WVXQQUWOKUZIEG-VEVYYDQMSA-N Pro-Thr-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O WVXQQUWOKUZIEG-VEVYYDQMSA-N 0.000 description 1
- AJJDPGVVNPUZCR-RHYQMDGZSA-N Pro-Thr-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@@H]1CCCN1)O AJJDPGVVNPUZCR-RHYQMDGZSA-N 0.000 description 1
- KHRLUIPIMIQFGT-AVGNSLFASA-N Pro-Val-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O KHRLUIPIMIQFGT-AVGNSLFASA-N 0.000 description 1
- 102000004245 Proteasome Endopeptidase Complex Human genes 0.000 description 1
- 108090000708 Proteasome Endopeptidase Complex Proteins 0.000 description 1
- 101710188315 Protein X Proteins 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- 101000702488 Rattus norvegicus High affinity cationic amino acid transporter 1 Proteins 0.000 description 1
- UEJYSALTSUZXFV-SRVKXCTJSA-N Rigin Chemical compound NCC(=O)N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCN=C(N)N)C(O)=O UEJYSALTSUZXFV-SRVKXCTJSA-N 0.000 description 1
- 241000220317 Rosa Species 0.000 description 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 1
- ZUGXSSFMTXKHJS-ZLUOBGJFSA-N Ser-Ala-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C)C(=O)N[C@@H](C)C(O)=O ZUGXSSFMTXKHJS-ZLUOBGJFSA-N 0.000 description 1
- YQHZVYJAGWMHES-ZLUOBGJFSA-N Ser-Ala-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O YQHZVYJAGWMHES-ZLUOBGJFSA-N 0.000 description 1
- QEDMOZUJTGEIBF-FXQIFTODSA-N Ser-Arg-Asp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(O)=O QEDMOZUJTGEIBF-FXQIFTODSA-N 0.000 description 1
- HZWAHWQZPSXNCB-BPUTZDHNSA-N Ser-Arg-Trp Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O HZWAHWQZPSXNCB-BPUTZDHNSA-N 0.000 description 1
- OOKCGAYXSNJBGQ-ZLUOBGJFSA-N Ser-Asn-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O OOKCGAYXSNJBGQ-ZLUOBGJFSA-N 0.000 description 1
- VAUMZJHYZQXZBQ-WHFBIAKZSA-N Ser-Asn-Gly Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O VAUMZJHYZQXZBQ-WHFBIAKZSA-N 0.000 description 1
- CTLVSHXLRVEILB-UBHSHLNASA-N Ser-Asn-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CO)N CTLVSHXLRVEILB-UBHSHLNASA-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
- MUJQWSAWLLRJCE-KATARQTJSA-N Ser-Leu-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MUJQWSAWLLRJCE-KATARQTJSA-N 0.000 description 1
- PPNPDKGQRFSCAC-CIUDSAMLSA-N Ser-Lys-Asp Chemical compound NCCCC[C@H](NC(=O)[C@@H](N)CO)C(=O)N[C@@H](CC(O)=O)C(O)=O PPNPDKGQRFSCAC-CIUDSAMLSA-N 0.000 description 1
- XUDRHBPSPAPDJP-SRVKXCTJSA-N Ser-Lys-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CO XUDRHBPSPAPDJP-SRVKXCTJSA-N 0.000 description 1
- RHAPJNVNWDBFQI-BQBZGAKWSA-N Ser-Pro-Gly Chemical compound OC[C@H](N)C(=O)N1CCC[C@H]1C(=O)NCC(O)=O RHAPJNVNWDBFQI-BQBZGAKWSA-N 0.000 description 1
- ZSDXEKUKQAKZFE-XAVMHZPKSA-N Ser-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CO)N)O ZSDXEKUKQAKZFE-XAVMHZPKSA-N 0.000 description 1
- ODRUTDLAONAVDV-IHRRRGAJSA-N Ser-Val-Tyr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O ODRUTDLAONAVDV-IHRRRGAJSA-N 0.000 description 1
- 101710172711 Structural protein Proteins 0.000 description 1
- 239000012505 Superdex™ Substances 0.000 description 1
- 102100021696 Syncytin-1 Human genes 0.000 description 1
- LMMDEZPNUTZJAY-GCJQMDKQSA-N Thr-Asp-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C)C(O)=O LMMDEZPNUTZJAY-GCJQMDKQSA-N 0.000 description 1
- KRPKYGOFYUNIGM-XVSYOHENSA-N Thr-Asp-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N)O KRPKYGOFYUNIGM-XVSYOHENSA-N 0.000 description 1
- MMTOHPRBJKEZHT-BWBBJGPYSA-N Thr-Cys-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(O)=O MMTOHPRBJKEZHT-BWBBJGPYSA-N 0.000 description 1
- LAFLAXHTDVNVEL-WDCWCFNPSA-N Thr-Gln-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CCCCN)C(=O)O)N)O LAFLAXHTDVNVEL-WDCWCFNPSA-N 0.000 description 1
- JMGJDTNUMAZNLX-RWRJDSDZSA-N Thr-Glu-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JMGJDTNUMAZNLX-RWRJDSDZSA-N 0.000 description 1
- HJOSVGCWOTYJFG-WDCWCFNPSA-N Thr-Glu-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N)O HJOSVGCWOTYJFG-WDCWCFNPSA-N 0.000 description 1
- KBBRNEDOYWMIJP-KYNKHSRBSA-N Thr-Gly-Thr Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)N[C@@H]([C@@H](C)O)C(=O)O)N)O KBBRNEDOYWMIJP-KYNKHSRBSA-N 0.000 description 1
- SXAGUVRFGJSFKC-ZEILLAHLSA-N Thr-His-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O SXAGUVRFGJSFKC-ZEILLAHLSA-N 0.000 description 1
- GXUWHVZYDAHFSV-FLBSBUHZSA-N Thr-Ile-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GXUWHVZYDAHFSV-FLBSBUHZSA-N 0.000 description 1
- WRQLCVIALDUQEQ-UNQGMJICSA-N Thr-Phe-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O WRQLCVIALDUQEQ-UNQGMJICSA-N 0.000 description 1
- ZESGVALRVJIVLZ-VFCFLDTKSA-N Thr-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@@H]1C(=O)O)N)O ZESGVALRVJIVLZ-VFCFLDTKSA-N 0.000 description 1
- 101710120037 Toxin CcdB Proteins 0.000 description 1
- MHNHRNHJMXAVHZ-AAEUAGOBSA-N Trp-Asn-Gly Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)NCC(=O)O)N MHNHRNHJMXAVHZ-AAEUAGOBSA-N 0.000 description 1
- XGFGVFMXDXALEV-XIRDDKMYSA-N Trp-Leu-Asn Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)N XGFGVFMXDXALEV-XIRDDKMYSA-N 0.000 description 1
- UQHPXCFAHVTWFU-BVSLBCMMSA-N Trp-Phe-Val Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O UQHPXCFAHVTWFU-BVSLBCMMSA-N 0.000 description 1
- SSSDKJMQMZTMJP-BVSLBCMMSA-N Trp-Tyr-Val Chemical compound C([C@@H](C(=O)N[C@@H](C(C)C)C(O)=O)NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)C1=CC=C(O)C=C1 SSSDKJMQMZTMJP-BVSLBCMMSA-N 0.000 description 1
- QYSBJAUCUKHSLU-JYJNAYRXSA-N Tyr-Arg-Val Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C(C)C)C(O)=O QYSBJAUCUKHSLU-JYJNAYRXSA-N 0.000 description 1
- OLYXUGBVBGSZDN-ACRUOGEOSA-N Tyr-Leu-Tyr Chemical compound C([C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=C(O)C=C1 OLYXUGBVBGSZDN-ACRUOGEOSA-N 0.000 description 1
- GZUIDWDVMWZSMI-KKUMJFAQSA-N Tyr-Lys-Cys Chemical compound NCCCC[C@@H](C(=O)N[C@@H](CS)C(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 GZUIDWDVMWZSMI-KKUMJFAQSA-N 0.000 description 1
- TYFLVOUZHQUBGM-IHRRRGAJSA-N Tyr-Ser-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 TYFLVOUZHQUBGM-IHRRRGAJSA-N 0.000 description 1
- FZSPNKUFROZBSG-ZKWXMUAHSA-N Val-Ala-Asp Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@H](C(O)=O)CC(O)=O FZSPNKUFROZBSG-ZKWXMUAHSA-N 0.000 description 1
- NMANTMWGQZASQN-QXEWZRGKSA-N Val-Arg-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N NMANTMWGQZASQN-QXEWZRGKSA-N 0.000 description 1
- VMRFIKXKOFNMHW-GUBZILKMSA-N Val-Arg-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N VMRFIKXKOFNMHW-GUBZILKMSA-N 0.000 description 1
- XLDYBRXERHITNH-QSFUFRPTSA-N Val-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)C(C)C XLDYBRXERHITNH-QSFUFRPTSA-N 0.000 description 1
- BMGOFDMKDVVGJG-NHCYSSNCSA-N Val-Asp-Lys Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BMGOFDMKDVVGJG-NHCYSSNCSA-N 0.000 description 1
- VVZDBPBZHLQPPB-XVKPBYJWSA-N Val-Glu-Gly Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)NCC(O)=O VVZDBPBZHLQPPB-XVKPBYJWSA-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
- PIFJAFRUVWZRKR-QMMMGPOBSA-N Val-Gly-Gly Chemical compound CC(C)[C@H]([NH3+])C(=O)NCC(=O)NCC([O-])=O PIFJAFRUVWZRKR-QMMMGPOBSA-N 0.000 description 1
- OACSGBOREVRSME-NHCYSSNCSA-N Val-His-Asn Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)N[C@@H](CC(N)=O)C(O)=O OACSGBOREVRSME-NHCYSSNCSA-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
- XXWBHOWRARMUOC-NHCYSSNCSA-N Val-Lys-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)N)C(=O)O)N XXWBHOWRARMUOC-NHCYSSNCSA-N 0.000 description 1
- VIKZGAUAKQZDOF-NRPADANISA-N Val-Ser-Glu Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O VIKZGAUAKQZDOF-NRPADANISA-N 0.000 description 1
- VHIZXDZMTDVFGX-DCAQKATOSA-N Val-Ser-Leu Chemical compound CC(C)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)N VHIZXDZMTDVFGX-DCAQKATOSA-N 0.000 description 1
- GBIUHAYJGWVNLN-UHFFFAOYSA-N Val-Ser-Pro Natural products CC(C)C(N)C(=O)NC(CO)C(=O)N1CCCC1C(O)=O GBIUHAYJGWVNLN-UHFFFAOYSA-N 0.000 description 1
- NZYNRRGJJVSSTJ-GUBZILKMSA-N Val-Ser-Val Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O NZYNRRGJJVSSTJ-GUBZILKMSA-N 0.000 description 1
- BGTDGENDNWGMDQ-KJEVXHAQSA-N Val-Tyr-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](C(C)C)N)O BGTDGENDNWGMDQ-KJEVXHAQSA-N 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 102220532017 WW domain-binding protein 11_K322A_mutation Human genes 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 238000001042 affinity chromatography Methods 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 108010076324 alanyl-glycyl-glycine Proteins 0.000 description 1
- 108010024078 alanyl-glycyl-serine Proteins 0.000 description 1
- 108010069020 alanyl-prolyl-glycine Proteins 0.000 description 1
- 108010005233 alanylglutamic acid Proteins 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000005875 antibody response Effects 0.000 description 1
- 230000010056 antibody-dependent cellular cytotoxicity Effects 0.000 description 1
- 208000037908 antibody-mediated disorder Diseases 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 108010052670 arginyl-glutamyl-glutamic acid Proteins 0.000 description 1
- 108010072041 arginyl-glycyl-aspartic acid Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 108010038633 aspartylglutamate Proteins 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 108010069495 cysteinyltyrosine Proteins 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 241001493065 dsRNA viruses Species 0.000 description 1
- 238000001976 enzyme digestion Methods 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 108010013768 glutamyl-aspartyl-proline Proteins 0.000 description 1
- 108010049041 glutamylalanine Proteins 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
- XBGGUPMXALFZOT-UHFFFAOYSA-N glycyl-L-tyrosine hemihydrate Natural products NCC(=O)NC(C(O)=O)CC1=CC=C(O)C=C1 XBGGUPMXALFZOT-UHFFFAOYSA-N 0.000 description 1
- 108010015792 glycyllysine Proteins 0.000 description 1
- 108010081551 glycylphenylalanine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000002163 immunogen Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 229940031551 inactivated vaccine Drugs 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 108010031424 isoleucyl-prolyl-proline Proteins 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- 108010027338 isoleucylcysteine Proteins 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 108010053037 kyotorphin Proteins 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 108010090333 leucyl-lysyl-proline Proteins 0.000 description 1
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 1
- 108010091871 leucylmethionine Proteins 0.000 description 1
- 230000003986 lysosome degradation Effects 0.000 description 1
- 108010003700 lysyl aspartic acid Proteins 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001404 mediated effect Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 238000007857 nested PCR Methods 0.000 description 1
- PGSADBUBUOPOJS-UHFFFAOYSA-N neutral red Chemical compound Cl.C1=C(C)C(N)=CC2=NC3=CC(N(C)C)=CC=C3N=C21 PGSADBUBUOPOJS-UHFFFAOYSA-N 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- 229940023146 nucleic acid vaccine Drugs 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 108010024654 phenylalanyl-prolyl-alanine Proteins 0.000 description 1
- 108010073101 phenylalanylleucine Proteins 0.000 description 1
- 108010051242 phenylalanylserine Proteins 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 108010070643 prolylglutamic acid Proteins 0.000 description 1
- 108010015796 prolylisoleucine Proteins 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- 108010048397 seryl-lysyl-leucine Proteins 0.000 description 1
- 230000000087 stabilizing effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 108010071097 threonyl-lysyl-proline Proteins 0.000 description 1
- 238000013519 translation Methods 0.000 description 1
- 108010052774 valyl-lysyl-glycyl-phenylalanyl-tyrosine Proteins 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 230000009385 viral infection Effects 0.000 description 1
- 229940023147 viral vector vaccine Drugs 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/005—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from viruses
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/12—Viral antigens
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/20011—Coronaviridae
- C12N2770/20022—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2770/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses positive-sense
- C12N2770/00011—Details
- C12N2770/20011—Coronaviridae
- C12N2770/20034—Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Molecular Biology (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Microbiology (AREA)
- Communicable Diseases (AREA)
- Oncology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Epidemiology (AREA)
- Immunology (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
本发明公开了一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用。所述融合蛋白包括融合表达的SARS‑CoV‑2病毒S蛋白的RBD区域和IgG抗体的Fc区域。本发明通过将SARS‑CoV‑2的棘突蛋白的RBD结构域与免疫球蛋白Fc进行融合表达获得重组融合蛋白,Fc的铰链区以及恒定区能通过二硫键将IgG抗体的两条重链相结合,实现RBD的二聚体,制备的重组融合蛋白可用于作为针对新型冠状病毒肺炎的重组蛋白疫苗。该重组蛋白疫苗可在小鼠体内诱导产生高滴度的特异性IgG抗体和中和抗体,抗体水平可维持3个月以上,该疫苗能同时诱导小鼠产生细胞免疫。
Description
技术领域
本发明涉及疫苗技术领域,特别是涉及一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用。
背景技术
冠状病毒属于尼多病毒目,冠状病毒科的一种具有包膜的单股正链的RNA病毒(+ss RNA),又可分为α、β、γ、δ属。其中,α、β属主要感染哺乳动物,而γ、δ属主要感染禽类,少数情况下也感染哺乳动物。新型冠状病毒肺炎(COVID-19)则是由β属的SARS-CoV-2病毒所引起。
SARS-CoV-2病毒其基因组编码四种结构蛋白,分别为棘突蛋白,包膜蛋白,膜蛋白以及核衣壳蛋白。其中,S蛋白的受体结合结构域(Receptor binding domain,RBD)通过结合宿主细胞膜上的血管紧张素转化酶2(ACE2)感染人体。在缺乏特效药的情况下,疫苗仍然是保护人群的最有效的防控手段。S蛋白在介导病毒感染的初期发挥重要作用,是产生中和抗体的主要区域。成熟的S蛋白是以三聚体形式构成病毒表面的“皇冠状”外形。S蛋白的膜远端S1结构域包括了RBD结构域,主要与宿主细胞膜上的ACE2受体结合。膜近端S2结构域包括了FP(Fusion peptide,融合肽)成分。S蛋白三聚体在未结合受体之前,2个RBD结构域处于“向下”状态,1个RBD结构域处于“向上”状态。在与受体结合之后,蛋白酶体识别并切割弗林蛋白酶切位点,继而导致S1结构脱落,接着触发S2亚基构象的改变,S2的FP成分暴露有利于病毒包膜和宿主细胞膜的融合,最终实现病毒包膜和宿主细胞膜的融合。当宿主感染SARS-CoV-2之后,体内会产生多种结合S蛋白的中和抗体,大部分靶向RBD结构域,少部分靶向NTD结构域。由于RBD结构域相比全长S蛋白,S1结构域,S2结构域,诱导产生的中和抗体能力显著强于其他结构,之前关于MERS和SARS的RBD重组蛋白已证明其作为重组蛋白疫苗开发的理想的候选抗原。
疫苗开发五大类型:减毒活疫苗、灭活疫苗、病毒载体疫苗、重组蛋白疫苗、核酸疫苗。
重组蛋白疫苗可细分为重组S蛋白疫苗、重组RBD疫苗和类病毒颗粒疫苗(VLPs)。这些重组蛋白可以通过不同的表达***表达,包括昆虫细胞、哺乳动物细胞、酵母、原核细胞以及植物。不同表达体系的差异体现在翻译后修饰。原核细胞不具有翻译后修饰的功能,因此丧失了对S蛋白或者RBD蛋白糖基化的修饰。但是,目前发现SARS-CoV-2的棘突蛋白表面存在许多糖基化修饰,也许这些糖基化修饰参与稳定RBD构,也可能作为是免疫原性的一部分。但是,有些糖基化修饰也可能掩蔽表位区域。因此,即便对于相同的重组蛋白疫苗,不同的表达体系也有可能导致其免疫原性的差异。一般来说,哺乳动物表达的RBD重组蛋白疫苗比昆虫和大肠杆菌中表达的RBD重组蛋白疫苗能够诱导更强的中和抗体反应。另一方面,重组蛋白疫苗通常仅包含中和表位产生的结构域,因此,这些疫苗大多数都含有全长S蛋白或者部分结构,比起S蛋白,RBD结构域可能缺乏了其他中和表位,比如靶向NTD区域。但是,全长S蛋白疫苗可能能够诱导不必要的免疫反应,导致抗体介导的疾病增强。RBD作为单体形式直接应用于疫苗,其免疫原性较弱。所以往往要结合适合的佐剂或者引入额外的蛋白形成二聚体以增加其免疫原性。
发明内容
本发明针对现有技术中存在的上述不足,提供了一种免疫效果更好、由SARS-CoV-2的棘突蛋白的RBD结构域与免疫球蛋白Fc进行融合表达的重组融合蛋白,可用于作为针对新型冠状病毒肺炎的重组蛋白疫苗。
一种融合蛋白,包括融合表达的SARS-CoV-2病毒S蛋白的RBD区域和IgG抗体的Fc区域。
优选的,RBD区域的氨基酸序列如SEQ ID No.1所示,该序列为棘突蛋白的330-583aa。
优选的,IgG抗体为人源IgG抗体或鼠源IgG抗体。更优选的,人源IgG抗体的Fc区域氨基酸序列如SEQ ID No.2所示;鼠源IgG抗体的Fc区域氨基酸序列如SEQ ID No.3所示。
优选的,RBD区域与Fc区域之间通过柔性连接接头进行连接,柔性连接接头的氨基酸序列为GGGGS。柔性连接接头的作用在于避免RBD结构域和Fc结构域可能出现的相互影响,当然除了Gly-Gly-Gly-Gly-Ser(GGGGS)序列外,其中Gly与Ser的数量、位置等均可以再调整,或者使用其他常用的柔性连接接头,柔性连接接头本身对RBD结构域和Fc结构域的结构没有影响。
本发明又提供了编码所述融合蛋白的基因。
优选的,编码RBD区域的序列如SEQ ID No.4所示,编码Fc区域的序列如SEQ IDNo.5或6所示。
所述的基因,还包括位于编码RBD区域的序列上游的信号肽编码序列以及位于信号肽编码序列上游的Kozak序列,信号肽编码序列如SEQ ID No.7所示,Kozak序列为ACCACC。Kozak序列是位于真核生物mRNA5’端帽子结构后面的一段核酸序列,它可以与翻译起始因子结合而介导含有5’帽子结构的mRNA翻译起始,用来增强真核基因的翻译效率的。信号肽用于在真核细胞表达***中做分泌表达,一般使用HEK293细胞进行表达的话,使用HEK293细胞的信号肽,如果用其他细胞表达***,就相应使用其他细胞对应的信号肽即可。
本发明还提供了所述融合蛋白在制备新型冠状病毒亚单位疫苗中的应用。
一种新型冠状病毒亚单位疫苗,包含所述融合蛋白。
本发明通过将SARS-CoV-2的棘突蛋白的RBD结构域与免疫球蛋白Fc进行融合表达获得重组融合蛋白,已知Fc的铰链区以及恒定区能通过二硫键将IgG抗体的两条重链相结合,因此,利用这一特性,将其应用于RBD二聚体蛋白的设计上,制备的重组融合蛋白可用于作为针对新型冠状病毒肺炎的重组蛋白疫苗。另外,Fc通过FcRn受体途径,可以将Fc融合药物从溶酶体的降解中拯救出来,从而有效延长药物的半衰期。
本发明制备的重组蛋白疫苗可在小鼠体内诱导产生高滴度的特异性IgG抗体和中和抗体,抗体水平可维持3个月以上,该疫苗能同时诱导小鼠产生细胞免疫。
附图说明
图1为连接鼠源Fc或者人源Fc的RBD重组蛋白疫苗示意图。
图2为分子筛纯化RBD重组蛋白疫苗的结果,其中,图A为RBD-hFc,图B为RBD-mFc。
图3为SDS-PAGE分析分子筛纯化后RBD重组蛋白疫苗的结果,其中,图A为RBD-hFc样品且为非还原性SDS-PAGE,图B为RBD-hFc样品且为还原性SDS-PAGE,图C为RBD-mFc样品且为非还原性SDS-PAGE,图D为RBD-mFc样品且为还原性SDS-PAGE,图E为S1-hFc样品且为还原性SDS-PAGE。
图4为SEC-HPLC分析分子筛纯化后RBD重组蛋白疫苗的结果,其中,图A为RBD-hFc,图B为RBD-mFc。
图5为VERO E6细胞检测RBD重组蛋白疫苗亲和力检测图,其中,图A为RBD-hFc,图B为RBD-mFc;图A和B中,左侧的峰为对照组,即一抗加入的PBS溶液。
图6为RBD重组蛋白免疫后IgG滴度检测的结果。其中对照组分别为AL(OH)3以及S1-hFc,其中,图A为RBD-hFc,图B为RBD-mFc,图C为S1-hFc。
图7为RBD重组蛋白免疫后中和抗体滴度检测的结果。其中对照组分别为AL(OH)3以及S1-hFc,其中,图A为RBD-hFc,图B为RBD-mFc,图C为S1-hFc。
图8为RBD重组蛋白免疫后细胞因子释放的结果,对照组分别为AL(OH)3以及S1-hFc,图A为IFN-γ释放量,图B为IL-4释放量,图C为IL-10释放量。
具体实施方式
实施例1:RBD重组蛋白的制备
(1)RBD序列源于SARS-CoV-2棘突蛋白330-583氨基酸范围。在本发明中,鼠源Fc来自于鼠源IgG1,人源Fc是在人源IgG1的基础上进行改造。人源Fc具体改造位点是在Asn297Ala以及Lys322Ala,对于疫苗而而言,Fc所介导的ADCC和CDC效应是多余的。为了增加疫苗在应用上的安全性,因此对这两个位点进行突变。
实验设计路线:先通过PCR分别扩增RBD片段(基因序列SEQ ID No.4,氨基酸序列SEQ ID No.1),人源IgG1修饰后的Fc片段(以下简称为hFc,基因序列SEQ ID No.5,氨基酸序列SEQ ID No.2),鼠源IgG1的Fc片段(以下简称为mFc,基因序列SEQ ID No.6,氨基酸序列SEQ ID No.3)。再通过重叠延伸PCR,将RBD与hFc以及mFc通过一段柔性连接区(Gly-Gly-Gly-Gly-Ser,基因序列SEQ ID No.10)进行拼接。第三步通过延伸PCR,在上游引入EcoR I酶切位点(GAATTC)、kozak序列(ACCACC)及HEK293细胞信号肽(氨基酸序列:MDMRVPAQLLGLLLLWFPGARC,基因序列SEQ ID No.7),此外在下游引入终止密码子(TGA)及XhoI酶切位点(GCGGCCGC)。最后将双酶切后的PCR产物连接于pcdna3.1真核表达载体中,并将连接产物转化DH5α大肠杆菌,挑选测序正确的单克隆并保菌。将上述重组质粒瞬转入人胚肾细胞(HEK293细胞)进行表达,将融合人源Fc的RBD重组蛋白命名为RBD-hFc,融合鼠源Fc的RBD重组蛋白命名为RBD-mFc。
RBD重组蛋白首先利用亲和层析法进行纯化:RBD-hFc利用protein A柱纯化,RBD-mFc利用protein G柱纯化。然后,将上一步纯化后的蛋白进一步利用分子筛(Superdex200Increase 10/300GL,GE)来纯化。由于受到Fc形成二聚体的能力,RBD-hFc产生两个邻近的峰,RBD-mFc主要产生一个峰(图2)。
(2)SDS-PAGE分析纯化产物如图3,RBD-hFc第一个峰的产物在还原条件下产生55KDa左右的条带,在非还原条件下产生170KDa左右的条带,因此峰1的产物符合理论蛋白分子量;RBD-mFc第二个峰的产物在还原条件下产生55KDa左右的条带,在非还原条件下产生170KDa左右的条带,因此峰2的产物符合理论蛋白分子量。同时,利用SEC-HPLC分析纯化产物(非还原条件下),如图4,说明RBD重组蛋白可以达到较高纯度。
实施例2:流式亲和力测定
(1)收集1×106个VERO E6细胞分别与RBD-hFc重组蛋白疫苗(10μg/ml),RBD-mFc重组蛋白疫苗(10μg/ml),并设置两组阴性对照(仅加入PBS)。在4℃孵育30min。
(2)PBS洗涤3次后,在孵育RBD-mFc的试管中加入FITC标记的山羊抗鼠的二抗(1:1000)200μl;在孵育RBD-mFc的试管中加入FITC标记的山羊抗鼠的二抗(1:1000)200μl;阴性对照组,分别加入FITC标记的山羊抗人的二抗(1:1000)200μl,FITC标记的山羊抗鼠的二抗(1:1000)200μl。4℃孵育30min之后,PBS洗涤3次后,用流式细胞仪检测。
(3)流式细胞仪检测RBD重组蛋白与VERO E6细胞(ACE2阳性)的亲和力主要通过二抗标记后的FITC的平均荧光强度来表达。
(4)如图5所示,RBD重组蛋白与VERO E6细胞的亲和力明显高于阴性对照组,因此Fc融合形式的RBD疫苗仍然保持其天然构象。
实施例3:重组蛋白免疫小鼠
选择健康的BALB/C雌性小鼠,8周龄(19-21g),小鼠随机分成7组,每组4只,分别为F,G,H,I,J,K,L共7组,具体如下:F组为接种Al(OH)3的佐剂组;G组为接种高剂量(8μg)RBD-hFc(RBD-hFcHigh),H组为接种低剂量(2μg)RBD-hFc(RBD-hFcLow);K组为接种高剂量(8μg)RBD-mFc(RBD-mFcHigh),L组为接种低剂量(2μg)RBD-mFc(RBD-mFcLow)。此外,为了比较RBD和S1两种Fc融合形式的重组蛋白疫苗的免疫原性差异,因此设置了融合人源Fc的S1重组蛋白疫苗作为对照,S1-hFc(购自北京义翘神州公司,货号40591-V02H)中S1结构域选择S蛋白16-685aa,而其中的Fc为人源IgG1(Asn297以及Lys322两个位点未突变)。
具体如下:I组为接种高剂量(8μg)S1-hFc(S1-hFcHigh),J组为接种低剂量(2μg)S1-hFc(S1-hFcLow)。
RBD以及对照的S1重组蛋白在结合Al(OH)3佐剂之后,采用后肢肌肉注射,注射总体积为200μl/只,每次接种时两侧后腿各注射一半剂量即100μl。初次免疫在第0天,免疫之后的第七天进行第二次的加强免疫。并且在免疫后1,2,3,4,6,8,10,12周采血,最后第91天处死小鼠并提取脾脏细胞。
实施例4:IgG特异性抗体滴度检测
为了检测RBD-hFc、RBD-mFc和S1-hFc融合蛋白的免疫原性,用高、低剂量的融合蛋白对BALB/C小鼠进行了两次免疫(方法同实施例3)。在免疫后1~12周分别进行采血。
先将50ng RBD(购自Genscript,China)蛋白在ELISA平板中4℃包被过夜。第二日,在每个孔中添加10%FBS,在37℃下在含有0.5%吐温-20的PBS中封闭1个小时。封闭之后,将稀释后的血清样本加入每个孔中,在37℃下孵育1个小时。接着,使用PBST溶液洗涤每个平板3次后,往每个孔中加入兔抗小鼠IgG HRP抗体,在37℃下孵育1个小时。同样,使用PBST溶液洗涤每个平板6次,并添加3,3',5,5'-四甲基联苯胺(TMB)底物,37℃下反应10分钟。用1%H2SO4停止反应,并在450nm处检测吸光度。OD450值大于背景值的2.1倍为阳性。
加强接种一周后,RBD-hFcHigh、RBD-hFcLow、RBD-mFcHigh、RBD-mFcLow、S1-hFcHigh和S1-hFcLow组的IgG抗体几何平均滴度(GMT)分别为21527、18102、15222、15222、3805和3200。6组IgG血清转化率均为100%。随后,IgG抗体滴度上升,在接种后3~4周达到高峰。RBD-mFcHigh组IgG抗体最高GMT超过10万。随着时间的推移,IgG抗体滴度略有下降,但在接种后12周仍维持在较高水平。RBD-hFc组和RBD-mFc组的IgG抗体水平均高于S1-hFc组。
实施例5:中和抗体滴度检测
首先用SARS-CoV-2病毒进行噬菌斑减少中和试验(PRNT)。来自免疫后小鼠的血清在56℃灭活0.5小时并连续稀释。将2倍稀释血清混合并与相同体积的病毒培养物(100TCID50)在37℃下培养1小时。将混合物添加到6孔板中的Vero E6细胞上,并在37℃下再培养1小时。然后,丢弃病毒培养物并用0.6%琼脂糖凝胶在病毒培养基中替换。孵育2天后,加入含0.1%中性红的第二层琼脂糖。一天后计算菌斑数量,并计算导致噬菌斑减少50%的血清稀释液(PRNT50)作为效价。
两次免疫后,除S1-hFcLow组的一只小鼠外,所有组在接种后两周都能检测到中和抗体滴度。中和抗体滴度在接种后4周达到高峰。RBD-hFcHigh、RBD-hFcLow、RBD-mFcHigh、RBD-mFcLow和S1-hFcHigh组的GMT分别为128、108、431、152和54。接种后12周后,RBD-hFcHigh组、RBD-hFcLow组、RBD-mFcHigh组和S1-hFcLow组GMT分别为最高值的35.4%、50%、35.4%和70.3%。而RBD-mFcLow组和S1-hFcLow组的中和抗体水平与峰值水平基本一致。结果表明,重组蛋白具有良好的免疫原性。
实施例6:细胞因子(IFN-γ、IL4以及IL10)检测
(1)取接种12周的RBD重组蛋白或AL佐剂免疫小鼠的血清标本。然后将各组小鼠等量的血清混合到一个样品中,使用Proteome ProfilerTM小鼠细胞因子阵列面板A对100μl血清样品进行分析。平均像素密度用Quantity One进行分析。
(2)从各组中随机抽取小鼠3只,处死后取脾脏细胞至组织培养皿。通过3ml注射器按压分离成单个细胞悬浮液。细胞在含有10%FBS的RPMI 1640培养基(5×105/孔)中培养,然后用SARS-CoV-2RBD(2μg/孔)刺激。在37℃,5%CO2中培养24小时后,使用小鼠酶联免疫斑点试剂盒测量产生IFN-γ的脾细胞。应用ChemiDoc-XRS+成像***对Spot-forming细胞(SFCs)进行成像,并用Quantity-One软件对相关数据进行统计分析。
为了评估RBD和S1融合蛋白的细胞免疫应答,从所有免疫小鼠中分离脾细胞并用RBD蛋白刺激。Elispot检测IFN-γ分泌水平明显高于对照组,其中RBD-mFcHigh组最高。用ELISA法检测RBD刺激后,脾细胞产生的IL-4和IL-10产生量。RBD-hFcHigh、RBD-mFcHigh和S1-hFcHigh组中脾细胞培养基的IL-4和IL-10浓度也显著高于对照组。三组IL-4浓度分别为18.6、19.9和16.3pg/ml,IL-10浓度分别为85.3、209和95.3pg/ml。这些结果表明,RBD重组蛋白能诱导小鼠产生细胞免疫应答。
序列表
<110> 浙江大学
浙江省疾病预防控制中心
<120> 一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用
<160> 10
<170> SIPOSequenceListing 1.0
<210> 1
<211> 254
<212> PRT
<213> 新型冠状病毒(SARS-CoV-2)
<400> 1
Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr
1 5 10 15
Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys
20 25 30
Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe
35 40 45
Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr
50 55 60
Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln
65 70 75 80
Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu
85 90 95
Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu
100 105 110
Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg
115 120 125
Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr
130 135 140
Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr
145 150 155 160
Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr
165 170 175
Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro
180 185 190
Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys
195 200 205
Cys Val Asn Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr
210 215 220
Glu Ser Asn Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile
225 230 235 240
Ala Asp Thr Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu
245 250
<210> 2
<211> 231
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Pro Lys Ser Cys Asp Lys Thr His Thr Cys Pro Pro Cys Pro Ala Pro
1 5 10 15
Glu Ala Ala Gly Gly Pro Ser Val Phe Leu Phe Pro Pro Lys Pro Lys
20 25 30
Asp Thr Leu Met Ile Ser Arg Thr Pro Glu Val Thr Cys Val Val Val
35 40 45
Asp Val Ser His Glu Asp Pro Glu Val Lys Phe Asn Trp Tyr Val Asp
50 55 60
Gly Val Glu Val His Asn Ala Lys Thr Lys Pro Arg Glu Glu Gln Tyr
65 70 75 80
Ala Ser Thr Tyr Arg Val Val Ser Val Leu Thr Val Leu His Gln Asp
85 90 95
Trp Leu Asn Gly Lys Glu Tyr Lys Cys Ala Val Ser Asn Lys Ala Leu
100 105 110
Gly Ala Pro Ile Glu Lys Thr Ile Ser Lys Ala Lys Gly Gln Pro Arg
115 120 125
Glu Pro Gln Val Tyr Thr Leu Pro Pro Ser Arg Asp Glu Leu Thr Lys
130 135 140
Asn Gln Val Ser Leu Thr Cys Leu Val Lys Gly Phe Tyr Pro Ser Asp
145 150 155 160
Ile Ala Val Glu Trp Glu Ser Asn Gly Gln Pro Glu Asn Asn Tyr Lys
165 170 175
Thr Thr Pro Pro Val Leu Asp Ser Asp Gly Ser Phe Phe Leu Tyr Ser
180 185 190
Lys Leu Thr Val Asp Lys Ser Arg Trp Gln Gln Gly Asn Val Phe Ser
195 200 205
Cys Ser Val Met His Glu Ala Leu His Asn His Tyr Thr Gln Lys Ser
210 215 220
Leu Ser Leu Ser Pro Gly Lys
225 230
<210> 3
<211> 225
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Val Arg Ser Gly Cys Lys Pro Cys Ile Cys Thr Val Pro Glu Val Ser
1 5 10 15
Ser Val Phe Ile Phe Pro Pro Lys Pro Lys Asp Val Leu Thr Ile Thr
20 25 30
Leu Thr Pro Lys Val Thr Cys Val Val Val Asp Ile Ser Lys Asp Asp
35 40 45
Pro Glu Val Gln Phe Ser Trp Phe Val Asp Asp Val Glu Val His Thr
50 55 60
Ala Gln Thr Gln Pro Arg Glu Glu Gln Phe Asn Ser Thr Phe Arg Ser
65 70 75 80
Val Ser Glu Leu Pro Ile Met His Gln Asp Trp Leu Asn Gly Lys Glu
85 90 95
Phe Lys Cys Arg Val Asn Ser Ala Ala Phe Pro Ala Pro Ile Glu Lys
100 105 110
Thr Ile Ser Lys Thr Lys Gly Arg Pro Lys Ala Pro Gln Val Tyr Thr
115 120 125
Ile Pro Pro Pro Lys Glu Gln Met Ala Lys Asp Lys Val Ser Leu Thr
130 135 140
Cys Met Ile Thr Asp Phe Phe Pro Glu Asp Ile Thr Val Glu Trp Gln
145 150 155 160
Trp Asn Gly Gln Pro Ala Glu Asn Tyr Lys Asn Thr Gln Pro Ile Met
165 170 175
Asp Thr Asp Gly Ser Tyr Phe Val Tyr Ser Lys Leu Asn Val Gln Lys
180 185 190
Ser Asn Trp Glu Ala Gly Asn Thr Phe Thr Cys Ser Val Leu His Glu
195 200 205
Gly Leu His Asn His His Thr Glu Lys Ser Leu Ser His Ser Pro Gly
210 215 220
Lys
225
<210> 4
<211> 762
<212> DNA
<213> 新型冠状病毒(SARS-CoV-2)
<400> 4
cctaacatta ccaacctctg cccatttgga gaggtgttta acgccacccg gttcgccagc 60
gtgtacgcct ggaaccggaa gaggatcagc aactgcgtgg ccgactacag cgtgctgtac 120
aacagcgcct ccttcagcac cttcaagtgc tacggggtga gccccacaaa gctgaacgat 180
ctgtgcttca ccaacgtata cgccgatagc ttcgtgatcc ggggggatga ggtgaggcag 240
atcgcccccg gccagacagg caagatcgcc gattacaact acaagctgcc cgatgacttc 300
accggctgcg tgatcgcctg gaacagcaac aacctggact ccaaggtggg cggcaactac 360
aactacctgt accgcctgtt caggaagtcc aacctgaagc cttttgagag ggatatcagc 420
acagagatct accaggccgg ctccacaccc tgcaacggcg tggaggggtt caactgctac 480
ttccccctgc agagctatgg cttccagccc acaaacgggg tggggtacca gccctacagg 540
gtggtggtgc tgagcttcga gctgctgcac gcccccgcca cagtgtgcgg gcccaagaag 600
tccaccaacc tggtgaaaaa caagtgcgtg aacttcaact tcaacgggct gacagggacc 660
ggcgtgctga cagagagcaa caagaagttc ctgcccttcc agcagttcgg gcgggatatc 720
gccgacacca cagacgccgt gagggacccc cagacactgg ag 762
<210> 5
<211> 696
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 5
cccaaatctt gtgacaaaac tcacacatgc ccaccgtgcc cagcacctga agccgctggg 60
ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat ctcccggacc 120
cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt caagttcaac 180
tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga ggagcagtac 240
gccagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg gctgaatggc 300
aaggagtaca agtgcgccgt ctccaacaaa gccctcggag cccccatcga gaaaaccatc 360
tccaaagcca aagggcagcc ccgagaacca caggtgtaca ccctgccccc atcccgggat 420
gagctgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta tcccagcgac 480
atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac cacgcctccc 540
gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga caagagcagg 600
tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca caaccactac 660
acgcagaaga gcctctccct gtctccgggt aaatga 696
<210> 6
<211> 678
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 6
gttagatctg gttgtaagcc ttgcatatgt acagtcccag aagtatcatc tgtcttcatc 60
ttccccccaa agcccaagga tgtgctcacc attactctga ctcctaaggt cacgtgtgtt 120
gtggtagaca tcagcaagga tgatcccgag gtccagttca gctggtttgt agatgatgtg 180
gaggtgcaca cagctcagac gcaaccccgg gaggagcagt tcaacagcac tttccgctca 240
gtcagtgaac ttcccatcat gcaccaggac tggctcaatg gcaaggagtt caaatgcagg 300
gtcaacagtg cagctttccc tgcccccatc gagaaaacca tctccaaaac caaaggcaga 360
ccgaaggctc cacaggtgta caccattcca cctcccaagg agcagatggc caaggataaa 420
gtcagtctga cctgcatgat aacagacttc ttccctgaag acattactgt ggagtggcag 480
tggaatgggc agccagcgga gaactacaag aacactcagc ccatcatgga cacagatggc 540
tcttacttcg tctacagcaa gctcaatgtg cagaagagca actgggaggc aggaaatact 600
ttcacctgct ctgtgttaca tgagggcctg cacaaccacc atactgagaa gagcctctcc 660
cactctcctg gtaaatga 678
<210> 7
<211> 66
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 7
atggatatga gggtgcctgc ccagctgctg ggactgctcc tgctgtggtt tcccggcgcc 60
agatgc 66
<210> 8
<211> 22
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 8
Met Asp Met Arg Val Pro Ala Gln Leu Leu Gly Leu Leu Leu Leu Trp
1 5 10 15
Phe Pro Gly Ala Arg Cys
20
<210> 9
<211> 5
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 9
Gly Gly Gly Gly Ser
1 5
<210> 10
<211> 15
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
ggaggaggcg gcagc 15
Claims (10)
1.一种融合蛋白,包括融合表达的SARS-CoV-2病毒S蛋白的RBD区域和IgG抗体的Fc区域。
2.根据权利要求1所述的融合蛋白,其特征在于,RBD区域的氨基酸序列如SEQ ID No.1所示。
3.根据权利要求1所述的融合蛋白,其特征在于,IgG抗体为人源IgG抗体或鼠源IgG抗体。
4.根据权利要求3所述的融合蛋白,其特征在于,人源IgG抗体的Fc区域氨基酸序列如SEQ ID No.2所示;鼠源IgG抗体的Fc区域氨基酸序列如SEQ ID No.3所示。
5.根据权利要求1所述的融合蛋白,其特征在于,RBD区域与Fc区域之间通过柔性连接接头进行连接,柔性连接接头的氨基酸序列为GGGGS。
6.编码权利要求1~5任一所述融合蛋白的基因。
7.根据权利要求6所述的基因,其特征在于,编码RBD区域的序列如SEQ ID No.4所示,编码Fc区域的序列如SEQ ID No.5或6所示。
8.根据权利要求7所述的基因,其特征在于,还包括位于编码RBD区域的序列上游的信号肽编码序列以及位于信号肽编码序列上游的Kozak序列,信号肽编码序列如SEQ ID No.7所示,Kozak序列为ACCACC。
9.权利要求1~5任一所述融合蛋白在制备新型冠状病毒亚单位疫苗中的应用。
10.一种新型冠状病毒亚单位疫苗,其特征在于,包含权利要求1~5任一所述融合蛋白。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110180894.7A CN112851824B (zh) | 2021-02-09 | 一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110180894.7A CN112851824B (zh) | 2021-02-09 | 一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112851824A true CN112851824A (zh) | 2021-05-28 |
CN112851824B CN112851824B (zh) | 2024-08-02 |
Family
ID=
Cited By (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113773372A (zh) * | 2021-08-15 | 2021-12-10 | 北京科兴中维生物技术有限公司 | 重组蛋白及其制备方法和应用 |
CN113912738A (zh) * | 2021-07-19 | 2022-01-11 | 华中农业大学 | 一种猫新型冠状病毒亚单位疫苗及其制备方法及应用 |
CN114014940A (zh) * | 2021-11-25 | 2022-02-08 | 华兰基因工程有限公司 | 一种2019-nCoV表面蛋白受体结合区融合蛋白制备方法 |
CN114196701A (zh) * | 2021-11-17 | 2022-03-18 | 浙江迪福润丝生物科技有限公司 | 一种sars-cov-2的二价重组新城疫病毒载体及相应疫苗株和制备方法 |
CN114574502A (zh) * | 2022-04-11 | 2022-06-03 | 四川大学 | 一种以复制缺陷腺相关病毒为载体的新型冠状病毒疫苗 |
CN116023511A (zh) * | 2023-02-03 | 2023-04-28 | 上海易安飞玛生物技术有限公司 | 一种融合蛋白及其在制备SARS-CoV-2疫苗中的应用 |
Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017036905A1 (en) * | 2015-09-04 | 2017-03-09 | Ucb Biopharma Sprl | Multimeric proteins which bind to human fc-receptors |
CN111333704A (zh) * | 2020-02-24 | 2020-06-26 | 军事科学院军事医学研究院微生物流行病研究所 | 新型冠状病毒covid-19疫苗、制备方法及其应用 |
CN111533809A (zh) * | 2020-04-21 | 2020-08-14 | 中国科学院武汉病毒研究所 | 针对新型冠状病毒的亚单位疫苗及应用 |
CN111662389A (zh) * | 2020-06-05 | 2020-09-15 | 广州中医药大学(广州中医药研究院) | 一种SARS-CoV-2的融合蛋白及其疫苗组合物 |
CN111926040A (zh) * | 2020-10-12 | 2020-11-13 | 天津中逸安健生物科技有限公司 | 一种新型冠状病毒rbd核苷酸序列、优化方法与应用 |
CN111939250A (zh) * | 2020-08-17 | 2020-11-17 | 郑州大学 | 一种预防covid-19的新型疫苗及其制备方法 |
CN112225814A (zh) * | 2020-09-29 | 2021-01-15 | 东莞博盛生物科技有限公司 | 一种新型冠状病毒rbd融合蛋白亚单位疫苗及其制备方法和应用 |
CN112300251A (zh) * | 2020-02-24 | 2021-02-02 | 四川大学 | 抗SARS-CoV-2感染的蛋白及疫苗 |
Patent Citations (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2017036905A1 (en) * | 2015-09-04 | 2017-03-09 | Ucb Biopharma Sprl | Multimeric proteins which bind to human fc-receptors |
CN111333704A (zh) * | 2020-02-24 | 2020-06-26 | 军事科学院军事医学研究院微生物流行病研究所 | 新型冠状病毒covid-19疫苗、制备方法及其应用 |
CN112300251A (zh) * | 2020-02-24 | 2021-02-02 | 四川大学 | 抗SARS-CoV-2感染的蛋白及疫苗 |
CN111533809A (zh) * | 2020-04-21 | 2020-08-14 | 中国科学院武汉病毒研究所 | 针对新型冠状病毒的亚单位疫苗及应用 |
CN111662389A (zh) * | 2020-06-05 | 2020-09-15 | 广州中医药大学(广州中医药研究院) | 一种SARS-CoV-2的融合蛋白及其疫苗组合物 |
CN111939250A (zh) * | 2020-08-17 | 2020-11-17 | 郑州大学 | 一种预防covid-19的新型疫苗及其制备方法 |
CN112225814A (zh) * | 2020-09-29 | 2021-01-15 | 东莞博盛生物科技有限公司 | 一种新型冠状病毒rbd融合蛋白亚单位疫苗及其制备方法和应用 |
CN111926040A (zh) * | 2020-10-12 | 2020-11-13 | 天津中逸安健生物科技有限公司 | 一种新型冠状病毒rbd核苷酸序列、优化方法与应用 |
Non-Patent Citations (1)
Title |
---|
LINLIN WANG ET AL: "Silencing Fc domains in T Cell–Engaging Bispecific Antibodies Improves T-Cell Trafficking and Antitumor Potency", CANCER IMMUNOL RES, vol. 7, no. 12, 15 October 2019 (2019-10-15) * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113912738A (zh) * | 2021-07-19 | 2022-01-11 | 华中农业大学 | 一种猫新型冠状病毒亚单位疫苗及其制备方法及应用 |
CN113773372A (zh) * | 2021-08-15 | 2021-12-10 | 北京科兴中维生物技术有限公司 | 重组蛋白及其制备方法和应用 |
CN114196701A (zh) * | 2021-11-17 | 2022-03-18 | 浙江迪福润丝生物科技有限公司 | 一种sars-cov-2的二价重组新城疫病毒载体及相应疫苗株和制备方法 |
CN114196701B (zh) * | 2021-11-17 | 2023-10-10 | 浙江迪福润丝生物科技有限公司 | 一种sars-cov-2的二价重组新城疫病毒载体及相应疫苗株和制备方法 |
CN114014940A (zh) * | 2021-11-25 | 2022-02-08 | 华兰基因工程有限公司 | 一种2019-nCoV表面蛋白受体结合区融合蛋白制备方法 |
CN114574502A (zh) * | 2022-04-11 | 2022-06-03 | 四川大学 | 一种以复制缺陷腺相关病毒为载体的新型冠状病毒疫苗 |
CN116023511A (zh) * | 2023-02-03 | 2023-04-28 | 上海易安飞玛生物技术有限公司 | 一种融合蛋白及其在制备SARS-CoV-2疫苗中的应用 |
CN116023511B (zh) * | 2023-02-03 | 2024-04-09 | 北京强本生物技术有限公司 | 一种融合蛋白及其在制备SARS-CoV-2疫苗中的应用 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN109666070B (zh) | 单克隆抗体mers-4v2及其编码基因和应用 | |
AU2020201853B2 (en) | DNA antibody constructs and method of using same | |
Earnest et al. | Neutralizing antibodies against Mayaro virus require Fc effector functions for protective activity | |
CN107090029B (zh) | 可溶性 pd-1变体、融合构建体及其用途 | |
CA2841376C (en) | N-terminal deleted gp120 immunogens | |
KR20200103681A (ko) | Il-2 개변체 | |
AU2021201586A1 (en) | DNA antibody constructs and method of using same | |
WO2013006244A1 (en) | Bispecific monoclonal antibody therapeutics against west nile virus with improved cns penetration | |
Hua et al. | Engineering broadly neutralizing antibodies for HIV prevention and therapy | |
CN108530535A (zh) | 结合肽聚糖识别蛋白1的抗体 | |
CN108277234A (zh) | 封闭cd47并激发抗肿瘤免疫的重组溶瘤腺病毒及其用途 | |
CN110746503B (zh) | 抗H7N9全人源单克隆抗体hIg311及其制备方法与应用 | |
Zaharatos et al. | HIV-1 and influenza antigens synthetically linked to IgG2a Fc elicit superior humoral responses compared to unmodified antigens in mice | |
CN112851824B (zh) | 一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用 | |
CN111434685B (zh) | 抗h7n9全人源单克隆抗体9i17及其制备方法与应用 | |
CN111434681B (zh) | 抗h7n9全人源单克隆抗体6j15及其制备方法与应用 | |
CN109536476A (zh) | 具备透明质酸酶活性的靶向融合蛋白、制备方法及用途 | |
CN111434684B (zh) | 抗h7n9全人源单克隆抗体5q2及其制备方法与应用 | |
CN111320686B (zh) | 抗h7n9全人源单克隆抗体2g3及其制备方法与应用 | |
CN112851824A (zh) | 一种融合蛋白及其在制备新型冠状病毒亚单位疫苗中的应用 | |
CN114106184B (zh) | 抗ca125抗原vhh结构域及含有其的双特异性抗体 | |
CN104610453A (zh) | 一类抗her2双靶向抗体、其制备方法及用途 | |
CN113698487A (zh) | 抗人ace2单克隆抗体及其应用 | |
CN1216913C (zh) | 人白细胞介素-17受体样分子融合蛋白及其编码基因与表达细胞系 | |
CN109836494A (zh) | 一种靶向cd20的单链抗体、嵌合抗原受体t细胞及其制备方法和应用 |
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 |