CN114805603B - 一种ctb/ltb多肽与vp8的融合蛋白及轮状病毒疫苗 - Google Patents
一种ctb/ltb多肽与vp8的融合蛋白及轮状病毒疫苗 Download PDFInfo
- Publication number
- CN114805603B CN114805603B CN202210355800.XA CN202210355800A CN114805603B CN 114805603 B CN114805603 B CN 114805603B CN 202210355800 A CN202210355800 A CN 202210355800A CN 114805603 B CN114805603 B CN 114805603B
- Authority
- CN
- China
- Prior art keywords
- ctb
- seq
- asn
- thr
- amino acid
- 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.)
- Active
Links
- 108020001507 fusion proteins Proteins 0.000 title claims abstract description 33
- 102000037865 fusion proteins Human genes 0.000 title claims abstract description 33
- 229940124859 Rotavirus vaccine Drugs 0.000 title claims description 10
- 125000003275 alpha amino acid group Chemical group 0.000 claims abstract description 35
- 239000002671 adjuvant Substances 0.000 claims description 15
- 108090000765 processed proteins & peptides Proteins 0.000 abstract description 41
- 229920001184 polypeptide Polymers 0.000 abstract description 39
- 102000004196 processed proteins & peptides Human genes 0.000 abstract description 39
- 108090000623 proteins and genes Proteins 0.000 abstract description 36
- 102000004169 proteins and genes Human genes 0.000 abstract description 34
- 241000702670 Rotavirus Species 0.000 abstract description 18
- 239000012634 fragment Substances 0.000 abstract description 16
- 101150006264 ctb-1 gene Proteins 0.000 abstract description 15
- 241000617996 Human rotavirus Species 0.000 abstract description 4
- 230000005847 immunogenicity Effects 0.000 abstract description 4
- 238000000746 purification Methods 0.000 abstract description 4
- 230000004927 fusion Effects 0.000 abstract description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 3
- 108700021730 rotavirus NS35 Proteins 0.000 abstract description 2
- 108091028043 Nucleic acid sequence Proteins 0.000 description 19
- 101000867232 Escherichia coli Heat-stable enterotoxin II Proteins 0.000 description 17
- 230000003053 immunization Effects 0.000 description 14
- 238000002649 immunization Methods 0.000 description 14
- 229960005486 vaccine Drugs 0.000 description 14
- 150000007523 nucleic acids Chemical group 0.000 description 12
- 206010008631 Cholera Diseases 0.000 description 11
- 241000699670 Mus sp. Species 0.000 description 11
- 108020004414 DNA Proteins 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- 210000004369 blood Anatomy 0.000 description 6
- 239000008280 blood Substances 0.000 description 6
- RAXXELZNTBOGNW-UHFFFAOYSA-N imidazole Natural products C1=CNC=N1 RAXXELZNTBOGNW-UHFFFAOYSA-N 0.000 description 6
- 206010012735 Diarrhoea Diseases 0.000 description 5
- 108090000978 Interleukin-4 Proteins 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 230000001900 immune effect Effects 0.000 description 5
- 241000588724 Escherichia coli Species 0.000 description 4
- 102100037850 Interferon gamma Human genes 0.000 description 4
- 108010074328 Interferon-gamma Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 210000001744 T-lymphocyte Anatomy 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 108010043322 lysyl-tryptophyl-alpha-lysine Proteins 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 108020004707 nucleic acids Proteins 0.000 description 4
- 102000039446 nucleic acids Human genes 0.000 description 4
- VWADICJNCPFKJS-ZLUOBGJFSA-N Asn-Ser-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O VWADICJNCPFKJS-ZLUOBGJFSA-N 0.000 description 3
- 101100454807 Caenorhabditis elegans lgg-1 gene Proteins 0.000 description 3
- 241000283707 Capra Species 0.000 description 3
- MIIGESVJEBDJMP-FHWLQOOXSA-N Glu-Phe-Tyr Chemical compound C([C@H](NC(=O)[C@H](CCC(O)=O)N)C(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(O)=O)C1=CC=CC=C1 MIIGESVJEBDJMP-FHWLQOOXSA-N 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- XQJCEKXQUJQNNK-ZLUOBGJFSA-N Ser-Ser-Ser Chemical compound OC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O XQJCEKXQUJQNNK-ZLUOBGJFSA-N 0.000 description 3
- SQHKXWODKJDZRC-LKXGYXEUSA-N Ser-Thr-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(O)=O SQHKXWODKJDZRC-LKXGYXEUSA-N 0.000 description 3
- 210000004241 Th2 cell Anatomy 0.000 description 3
- GARULAKWZGFIKC-RWRJDSDZSA-N Thr-Gln-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O GARULAKWZGFIKC-RWRJDSDZSA-N 0.000 description 3
- PFMSJVIPEZMKSC-DZKIICNBSA-N Val-Tyr-Glu Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N PFMSJVIPEZMKSC-DZKIICNBSA-N 0.000 description 3
- 108010069205 aspartyl-phenylalanine Proteins 0.000 description 3
- 230000004069 differentiation Effects 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 108010062266 glycyl-glycyl-argininal Proteins 0.000 description 3
- 108010047926 leucyl-lysyl-tyrosine Proteins 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000013612 plasmid Substances 0.000 description 3
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 3
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 2
- NXSFUECZFORGOG-CIUDSAMLSA-N Ala-Asn-Leu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NXSFUECZFORGOG-CIUDSAMLSA-N 0.000 description 2
- SLQQPJBDBVPVQV-JYJNAYRXSA-N Arg-Phe-Val Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(O)=O SLQQPJBDBVPVQV-JYJNAYRXSA-N 0.000 description 2
- RYQSYXFGFOTJDJ-RHYQMDGZSA-N Arg-Thr-Leu Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O RYQSYXFGFOTJDJ-RHYQMDGZSA-N 0.000 description 2
- IOTKDTZEEBZNCM-UGYAYLCHSA-N Asn-Asn-Ile Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOTKDTZEEBZNCM-UGYAYLCHSA-N 0.000 description 2
- JZDZLBJVYWIIQU-AVGNSLFASA-N Asn-Glu-Tyr Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O JZDZLBJVYWIIQU-AVGNSLFASA-N 0.000 description 2
- QUMKPKWYDVMGNT-NUMRIWBASA-N Asn-Thr-Gln Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O QUMKPKWYDVMGNT-NUMRIWBASA-N 0.000 description 2
- POTCZYQVVNXUIG-BQBZGAKWSA-N Asp-Gly-Pro Chemical compound OC(=O)C[C@H](N)C(=O)NCC(=O)N1CCC[C@H]1C(O)=O POTCZYQVVNXUIG-BQBZGAKWSA-N 0.000 description 2
- HTSSXFASOUSJQG-IHPCNDPISA-N Asp-Tyr-Trp Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O HTSSXFASOUSJQG-IHPCNDPISA-N 0.000 description 2
- 241000894006 Bacteria Species 0.000 description 2
- 108090000695 Cytokines Proteins 0.000 description 2
- 102000004127 Cytokines Human genes 0.000 description 2
- 238000002965 ELISA Methods 0.000 description 2
- AQPZYBSRDRZBAG-AVGNSLFASA-N Gln-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](CCC(=O)N)N AQPZYBSRDRZBAG-AVGNSLFASA-N 0.000 description 2
- QOXDAWODGSIDDI-GUBZILKMSA-N Glu-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(=O)O)N QOXDAWODGSIDDI-GUBZILKMSA-N 0.000 description 2
- XOEKMEAOMXMURD-JYJNAYRXSA-N Glu-Tyr-His Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O XOEKMEAOMXMURD-JYJNAYRXSA-N 0.000 description 2
- WKJKBELXHCTHIJ-WPRPVWTQSA-N Gly-Arg-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CCCN=C(N)N WKJKBELXHCTHIJ-WPRPVWTQSA-N 0.000 description 2
- QSVCIFZPGLOZGH-WDSKDSINSA-N Gly-Glu-Ser Chemical compound NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O QSVCIFZPGLOZGH-WDSKDSINSA-N 0.000 description 2
- MBOAPAXLTUSMQI-JHEQGTHGSA-N Gly-Glu-Thr Chemical compound [H]NCC(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MBOAPAXLTUSMQI-JHEQGTHGSA-N 0.000 description 2
- UESJMAMHDLEHGM-NHCYSSNCSA-N Gly-Ile-Leu Chemical compound NCC(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O UESJMAMHDLEHGM-NHCYSSNCSA-N 0.000 description 2
- SOEGEPHNZOISMT-BYPYZUCNSA-N Gly-Ser-Gly Chemical compound NCC(=O)N[C@@H](CO)C(=O)NCC(O)=O SOEGEPHNZOISMT-BYPYZUCNSA-N 0.000 description 2
- QYZYJFXHXYUZMZ-UGYAYLCHSA-N Ile-Asn-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N QYZYJFXHXYUZMZ-UGYAYLCHSA-N 0.000 description 2
- KEKTTYCXKGBAAL-VGDYDELISA-N Ile-His-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CO)C(=O)O)N KEKTTYCXKGBAAL-VGDYDELISA-N 0.000 description 2
- GAZGFPOZOLEYAJ-YTFOTSKYSA-N Ile-Leu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N GAZGFPOZOLEYAJ-YTFOTSKYSA-N 0.000 description 2
- UAELWXJFLZBKQS-WHOFXGATSA-N Ile-Phe-Gly Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](Cc1ccccc1)C(=O)NCC(O)=O UAELWXJFLZBKQS-WHOFXGATSA-N 0.000 description 2
- SVZFKLBRCYCIIY-CYDGBPFRSA-N Ile-Pro-Arg Chemical compound CC[C@H](C)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SVZFKLBRCYCIIY-CYDGBPFRSA-N 0.000 description 2
- LZDNBBYBDGBADK-UHFFFAOYSA-N L-valyl-L-tryptophan Natural products C1=CC=C2C(CC(NC(=O)C(N)C(C)C)C(O)=O)=CNC2=C1 LZDNBBYBDGBADK-UHFFFAOYSA-N 0.000 description 2
- IIKJNQWOQIWWMR-CIUDSAMLSA-N Leu-Cys-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CS)NC(=O)[C@H](CC(C)C)N IIKJNQWOQIWWMR-CIUDSAMLSA-N 0.000 description 2
- YUTZYVTZDVZBJJ-IHPCNDPISA-N Lys-Trp-Lys Chemical compound C1=CC=C2C(C[C@H](NC(=O)[C@@H](N)CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O)=CNC2=C1 YUTZYVTZDVZBJJ-IHPCNDPISA-N 0.000 description 2
- RQILLQOQXLZTCK-KBPBESRZSA-N Lys-Tyr-Gly Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)NCC(O)=O RQILLQOQXLZTCK-KBPBESRZSA-N 0.000 description 2
- XMBSYZWANAQXEV-UHFFFAOYSA-N N-alpha-L-glutamyl-L-phenylalanine Natural products OC(=O)CCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 XMBSYZWANAQXEV-UHFFFAOYSA-N 0.000 description 2
- 108010002311 N-glycylglutamic acid Proteins 0.000 description 2
- 108010066427 N-valyltryptophan Proteins 0.000 description 2
- JEGFCFLCRSJCMA-IHRRRGAJSA-N Phe-Arg-Ser Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)N[C@@H](CO)C(=O)O)N JEGFCFLCRSJCMA-IHRRRGAJSA-N 0.000 description 2
- KDYPMIZMXDECSU-JYJNAYRXSA-N Phe-Leu-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 KDYPMIZMXDECSU-JYJNAYRXSA-N 0.000 description 2
- LKRUQZQZMXMKEQ-SFJXLCSZSA-N Phe-Trp-Thr Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LKRUQZQZMXMKEQ-SFJXLCSZSA-N 0.000 description 2
- BTPAWKABYQMKKN-LKXGYXEUSA-N Ser-Asp-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O BTPAWKABYQMKKN-LKXGYXEUSA-N 0.000 description 2
- ZOHGLPQGEHSLPD-FXQIFTODSA-N Ser-Gln-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O ZOHGLPQGEHSLPD-FXQIFTODSA-N 0.000 description 2
- FZXOPYUEQGDGMS-ACZMJKKPSA-N Ser-Ser-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O FZXOPYUEQGDGMS-ACZMJKKPSA-N 0.000 description 2
- 210000000447 Th1 cell Anatomy 0.000 description 2
- TZKPNGDGUVREEB-FOHZUACHSA-N Thr-Asn-Gly Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O TZKPNGDGUVREEB-FOHZUACHSA-N 0.000 description 2
- UYTYTDMCDBPDSC-URLPEUOOSA-N Thr-Ile-Phe Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N UYTYTDMCDBPDSC-URLPEUOOSA-N 0.000 description 2
- GYUUYCIXELGTJS-MEYUZBJRSA-N Thr-Phe-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC2=CN=CN2)C(=O)O)N)O GYUUYCIXELGTJS-MEYUZBJRSA-N 0.000 description 2
- MROIJTGJGIDEEJ-RCWTZXSCSA-N Thr-Pro-Pro Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N1[C@H](C(O)=O)CCC1 MROIJTGJGIDEEJ-RCWTZXSCSA-N 0.000 description 2
- YRJOLUDFVAUXLI-GSSVUCPTSA-N Thr-Thr-Asp Chemical compound C[C@@H](O)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(O)=O YRJOLUDFVAUXLI-GSSVUCPTSA-N 0.000 description 2
- QJIODPFLAASXJC-JHYOHUSXSA-N Thr-Thr-Phe Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)N)O QJIODPFLAASXJC-JHYOHUSXSA-N 0.000 description 2
- CKKFTIQYURNSEI-IHRRRGAJSA-N Tyr-Asn-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 CKKFTIQYURNSEI-IHRRRGAJSA-N 0.000 description 2
- KSFXWENSJABBFI-ZKWXMUAHSA-N Val-Ser-Asn Chemical compound [H]N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O KSFXWENSJABBFI-ZKWXMUAHSA-N 0.000 description 2
- 108010005233 alanylglutamic acid Proteins 0.000 description 2
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 239000000427 antigen Substances 0.000 description 2
- 102000036639 antigens Human genes 0.000 description 2
- 108091007433 antigens Proteins 0.000 description 2
- 108010068380 arginylarginine Proteins 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 230000018044 dehydration Effects 0.000 description 2
- 238000006297 dehydration reaction Methods 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 108010079547 glutamylmethionine Proteins 0.000 description 2
- 108010050848 glycylleucine Proteins 0.000 description 2
- 108010037850 glycylvaline Proteins 0.000 description 2
- 230000028993 immune response Effects 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 238000004806 packaging method and process Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000035755 proliferation Effects 0.000 description 2
- 108010079317 prolyl-tyrosine Proteins 0.000 description 2
- 108010004914 prolylarginine Proteins 0.000 description 2
- 230000005180 public health Effects 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- ROLXPVQSRCPVGK-XDTLVQLUSA-N Ala-Glu-Tyr Chemical compound N[C@@H](C)C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O ROLXPVQSRCPVGK-XDTLVQLUSA-N 0.000 description 1
- CKLDHDOIYBVUNP-KBIXCLLPSA-N Ala-Ile-Glu Chemical compound [H]N[C@@H](C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(O)=O CKLDHDOIYBVUNP-KBIXCLLPSA-N 0.000 description 1
- KUFVXLQLDHJVOG-SHGPDSBTSA-N Ala-Thr-Thr Chemical compound C[C@H]([C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](C)N)O KUFVXLQLDHJVOG-SHGPDSBTSA-N 0.000 description 1
- OTOXOKCIIQLMFH-KZVJFYERSA-N Arg-Ala-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCN=C(N)N OTOXOKCIIQLMFH-KZVJFYERSA-N 0.000 description 1
- NABSCJGZKWSNHX-RCWTZXSCSA-N Arg-Arg-Thr Chemical compound NC(N)=NCCC[C@@H](C(=O)N[C@@H]([C@H](O)C)C(O)=O)NC(=O)[C@@H](N)CCCN=C(N)N NABSCJGZKWSNHX-RCWTZXSCSA-N 0.000 description 1
- MTANSHNQTWPZKP-KKUMJFAQSA-N Arg-Gln-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CCCN=C(N)N)N)O MTANSHNQTWPZKP-KKUMJFAQSA-N 0.000 description 1
- VRTWYUYCJGNFES-CIUDSAMLSA-N Arg-Ser-Gln Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(O)=O VRTWYUYCJGNFES-CIUDSAMLSA-N 0.000 description 1
- FRBAHXABMQXSJQ-FXQIFTODSA-N Arg-Ser-Ser Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O FRBAHXABMQXSJQ-FXQIFTODSA-N 0.000 description 1
- UTSMXMABBPFVJP-SZMVWBNQSA-N Arg-Val-Trp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)NC(=O)[C@H](CCCN=C(N)N)N UTSMXMABBPFVJP-SZMVWBNQSA-N 0.000 description 1
- GMRGSBAMMMVDGG-GUBZILKMSA-N Asn-Arg-Arg Chemical compound C(C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)CN=C(N)N GMRGSBAMMMVDGG-GUBZILKMSA-N 0.000 description 1
- PCKRJVZAQZWNKM-WHFBIAKZSA-N Asn-Asn-Gly Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O PCKRJVZAQZWNKM-WHFBIAKZSA-N 0.000 description 1
- QISZHYWZHJRDAO-CIUDSAMLSA-N Asn-Asp-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N QISZHYWZHJRDAO-CIUDSAMLSA-N 0.000 description 1
- VYLVOMUVLMGCRF-ZLUOBGJFSA-N Asn-Asp-Ser Chemical compound NC(=O)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O VYLVOMUVLMGCRF-ZLUOBGJFSA-N 0.000 description 1
- PAXHINASXXXILC-SRVKXCTJSA-N Asn-Asp-Tyr Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC(=O)N)N)O PAXHINASXXXILC-SRVKXCTJSA-N 0.000 description 1
- COUZKSSMBFADSB-AVGNSLFASA-N Asn-Glu-Phe Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC(=O)N)N COUZKSSMBFADSB-AVGNSLFASA-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
- OOWSBIOUKIUWLO-RCOVLWMOSA-N Asn-Gly-Val Chemical compound [H]N[C@@H](CC(N)=O)C(=O)NCC(=O)N[C@@H](C(C)C)C(O)=O OOWSBIOUKIUWLO-RCOVLWMOSA-N 0.000 description 1
- SEKBHZJLARBNPB-GHCJXIJMSA-N Asn-Ile-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CO)C(O)=O SEKBHZJLARBNPB-GHCJXIJMSA-N 0.000 description 1
- IBLAOXSULLECQZ-IUKAMOBKSA-N Asn-Ile-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CC(N)=O IBLAOXSULLECQZ-IUKAMOBKSA-N 0.000 description 1
- ZMUQQMGITUJQTI-CIUDSAMLSA-N Asn-Leu-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O ZMUQQMGITUJQTI-CIUDSAMLSA-N 0.000 description 1
- XFJKRRCWLTZIQA-XIRDDKMYSA-N Asn-Lys-Trp Chemical compound C1=CC=C2C(=C1)C(=CN2)C[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC(=O)N)N XFJKRRCWLTZIQA-XIRDDKMYSA-N 0.000 description 1
- REQUGIWGOGSOEZ-ZLUOBGJFSA-N Asn-Ser-Asn Chemical compound C([C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)C(=O)N REQUGIWGOGSOEZ-ZLUOBGJFSA-N 0.000 description 1
- QYRMBFWDSFGSFC-OLHMAJIHSA-N Asn-Thr-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N)O QYRMBFWDSFGSFC-OLHMAJIHSA-N 0.000 description 1
- PQKSVQSMTHPRIB-ZKWXMUAHSA-N Asn-Val-Ser Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CO)C(O)=O PQKSVQSMTHPRIB-ZKWXMUAHSA-N 0.000 description 1
- AITKTFCQOBRJTG-CIUDSAMLSA-N Asp-Leu-Cys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CC(=O)O)N AITKTFCQOBRJTG-CIUDSAMLSA-N 0.000 description 1
- HJCGDIGVVWETRO-ZPFDUUQYSA-N Asp-Lys-Ile Chemical compound CC[C@H](C)[C@H](NC(=O)[C@H](CCCCN)NC(=O)[C@@H](N)CC(O)=O)C(O)=O HJCGDIGVVWETRO-ZPFDUUQYSA-N 0.000 description 1
- XXAMCEGRCZQGEM-ZLUOBGJFSA-N Asp-Ser-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(O)=O XXAMCEGRCZQGEM-ZLUOBGJFSA-N 0.000 description 1
- MGSVBZIBCCKGCY-ZLUOBGJFSA-N Asp-Ser-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O MGSVBZIBCCKGCY-ZLUOBGJFSA-N 0.000 description 1
- 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
- 238000011725 BALB/c mouse Methods 0.000 description 1
- 208000001726 Classical Swine Fever Diseases 0.000 description 1
- 108020004705 Codon Proteins 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- GRNOCLDFUNCIDW-ACZMJKKPSA-N Cys-Ala-Glu Chemical compound C[C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)O)NC(=O)[C@H](CS)N GRNOCLDFUNCIDW-ACZMJKKPSA-N 0.000 description 1
- KLLFLHBKSJAUMZ-ACZMJKKPSA-N Cys-Asn-Glu Chemical compound C(CC(=O)O)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CS)N KLLFLHBKSJAUMZ-ACZMJKKPSA-N 0.000 description 1
- LKHMGNHQULEPFY-ACZMJKKPSA-N Cys-Ser-Glu Chemical compound SC[C@H](N)C(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCC(O)=O LKHMGNHQULEPFY-ACZMJKKPSA-N 0.000 description 1
- 108010041986 DNA Vaccines Proteins 0.000 description 1
- 229940021995 DNA vaccine Drugs 0.000 description 1
- PHZYLYASFWHLHJ-FXQIFTODSA-N Gln-Asn-Glu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O PHZYLYASFWHLHJ-FXQIFTODSA-N 0.000 description 1
- ZNZPKVQURDQFFS-FXQIFTODSA-N Gln-Glu-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O ZNZPKVQURDQFFS-FXQIFTODSA-N 0.000 description 1
- XITLYYAIPBBHPX-ZKWXMUAHSA-N Gln-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](N)CCC(N)=O XITLYYAIPBBHPX-ZKWXMUAHSA-N 0.000 description 1
- UWMDGPFFTKDUIY-HJGDQZAQSA-N Gln-Pro-Thr Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O UWMDGPFFTKDUIY-HJGDQZAQSA-N 0.000 description 1
- UEILCTONAMOGBR-RWRJDSDZSA-N Gln-Thr-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O UEILCTONAMOGBR-RWRJDSDZSA-N 0.000 description 1
- UQKVUFGUSVYJMQ-IRIUXVKKSA-N Gln-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](CCC(=O)N)N)O UQKVUFGUSVYJMQ-IRIUXVKKSA-N 0.000 description 1
- XEKAJTCACGEBOK-KKUMJFAQSA-N Glu-Met-Phe Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)O)NC(=O)[C@H](CCC(=O)O)N XEKAJTCACGEBOK-KKUMJFAQSA-N 0.000 description 1
- UMZHHILWZBFPGL-LOKLDPHHSA-N Glu-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O UMZHHILWZBFPGL-LOKLDPHHSA-N 0.000 description 1
- NTHIHAUEXVTXQG-KKUMJFAQSA-N Glu-Tyr-Arg Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)NC(=O)[C@H](CCC(=O)O)N)O NTHIHAUEXVTXQG-KKUMJFAQSA-N 0.000 description 1
- HJTSRYLPAYGEEC-SIUGBPQLSA-N Glu-Tyr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)NC(=O)[C@H](CCC(=O)O)N HJTSRYLPAYGEEC-SIUGBPQLSA-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
- HQRHFUYMGCHHJS-LURJTMIESA-N Gly-Gly-Arg Chemical compound NCC(=O)NCC(=O)N[C@H](C(O)=O)CCCN=C(N)N HQRHFUYMGCHHJS-LURJTMIESA-N 0.000 description 1
- JNGHLWWFPGIJER-STQMWFEESA-N Gly-Pro-Tyr Chemical compound NCC(=O)N1CCC[C@H]1C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 JNGHLWWFPGIJER-STQMWFEESA-N 0.000 description 1
- KSOBNUBCYHGUKH-UWVGGRQHSA-N Gly-Val-Val Chemical compound CC(C)[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)CN KSOBNUBCYHGUKH-UWVGGRQHSA-N 0.000 description 1
- HAPWZEVRQYGLSG-IUCAKERBSA-N His-Gly-Glu Chemical compound [H]N[C@@H](CC1=CNC=N1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O HAPWZEVRQYGLSG-IUCAKERBSA-N 0.000 description 1
- BDFCIKANUNMFGB-PMVVWTBXSA-N His-Gly-Thr Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CN=CN1 BDFCIKANUNMFGB-PMVVWTBXSA-N 0.000 description 1
- JMSONHOUHFDOJH-GUBZILKMSA-N His-Ser-Glu Chemical compound OC(=O)CC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC1=CN=CN1 JMSONHOUHFDOJH-GUBZILKMSA-N 0.000 description 1
- GYXDQXPCPASCNR-NHCYSSNCSA-N His-Val-Asn Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N GYXDQXPCPASCNR-NHCYSSNCSA-N 0.000 description 1
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 1
- 241000701044 Human gammaherpesvirus 4 Species 0.000 description 1
- HDODQNPMSHDXJT-GHCJXIJMSA-N Ile-Asn-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O HDODQNPMSHDXJT-GHCJXIJMSA-N 0.000 description 1
- LBRCLQMZAHRTLV-ZKWXMUAHSA-N Ile-Gly-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O LBRCLQMZAHRTLV-ZKWXMUAHSA-N 0.000 description 1
- UWLHDGMRWXHFFY-HPCHECBXSA-N Ile-Ile-Pro Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N1CCC[C@@H]1C(=O)O)N UWLHDGMRWXHFFY-HPCHECBXSA-N 0.000 description 1
- OUUCIIJSBIBCHB-ZPFDUUQYSA-N Ile-Leu-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O OUUCIIJSBIBCHB-ZPFDUUQYSA-N 0.000 description 1
- TVYWVSJGSHQWMT-AJNGGQMLSA-N Ile-Leu-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N TVYWVSJGSHQWMT-AJNGGQMLSA-N 0.000 description 1
- AGGIYSLVUKVOPT-HTFCKZLJSA-N Ile-Ser-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N AGGIYSLVUKVOPT-HTFCKZLJSA-N 0.000 description 1
- YCKPUHHMCFSUMD-IUKAMOBKSA-N Ile-Thr-Asp Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N YCKPUHHMCFSUMD-IUKAMOBKSA-N 0.000 description 1
- COWHUQXTSYTKQC-RWRJDSDZSA-N Ile-Thr-Glu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(=O)O)C(=O)O)N COWHUQXTSYTKQC-RWRJDSDZSA-N 0.000 description 1
- HJDZMPFEXINXLO-QPHKQPEJSA-N Ile-Thr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)N HJDZMPFEXINXLO-QPHKQPEJSA-N 0.000 description 1
- NGKPIPCGMLWHBX-WZLNRYEVSA-N Ile-Tyr-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)O)N NGKPIPCGMLWHBX-WZLNRYEVSA-N 0.000 description 1
- 208000005168 Intussusception Diseases 0.000 description 1
- HGCNKOLVKRAVHD-UHFFFAOYSA-N L-Met-L-Phe Natural products CSCCC(N)C(=O)NC(C(O)=O)CC1=CC=CC=C1 HGCNKOLVKRAVHD-UHFFFAOYSA-N 0.000 description 1
- DBVWMYGBVFCRBE-CIUDSAMLSA-N Leu-Asn-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O DBVWMYGBVFCRBE-CIUDSAMLSA-N 0.000 description 1
- ULXYQAJWJGLCNR-YUMQZZPRSA-N Leu-Asp-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)NCC(O)=O ULXYQAJWJGLCNR-YUMQZZPRSA-N 0.000 description 1
- PNUCWVAGVNLUMW-CIUDSAMLSA-N Leu-Cys-Ser Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CS)C(=O)N[C@@H](CO)C(O)=O PNUCWVAGVNLUMW-CIUDSAMLSA-N 0.000 description 1
- PRZVBIAOPFGAQF-SRVKXCTJSA-N Leu-Glu-Met Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCSC)C(O)=O PRZVBIAOPFGAQF-SRVKXCTJSA-N 0.000 description 1
- USLNHQZCDQJBOV-ZPFDUUQYSA-N Leu-Ile-Asn Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O USLNHQZCDQJBOV-ZPFDUUQYSA-N 0.000 description 1
- ONPJGOIVICHWBW-BZSNNMDCSA-N Leu-Lys-Tyr Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 ONPJGOIVICHWBW-BZSNNMDCSA-N 0.000 description 1
- ILDSIMPXNFWKLH-KATARQTJSA-N Leu-Thr-Ser Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(O)=O ILDSIMPXNFWKLH-KATARQTJSA-N 0.000 description 1
- XTONYTDATVADQH-CIUDSAMLSA-N Lys-Cys-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(O)=O XTONYTDATVADQH-CIUDSAMLSA-N 0.000 description 1
- PGBPWPTUOSCNLE-JYJNAYRXSA-N Lys-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](CCCCN)N PGBPWPTUOSCNLE-JYJNAYRXSA-N 0.000 description 1
- LNMKRJJLEFASGA-BZSNNMDCSA-N Lys-Phe-Leu Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(C)C)C(O)=O LNMKRJJLEFASGA-BZSNNMDCSA-N 0.000 description 1
- IUYCGMNKIZDRQI-BQBZGAKWSA-N Met-Gly-Ala Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@@H](C)C(O)=O IUYCGMNKIZDRQI-BQBZGAKWSA-N 0.000 description 1
- WXJXYMFUTRXRGO-UWVGGRQHSA-N Met-His-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@H](C(=O)NCC(O)=O)CC1=CNC=N1 WXJXYMFUTRXRGO-UWVGGRQHSA-N 0.000 description 1
- WPTDJKDGICUFCP-XUXIUFHCSA-N Met-Ile-Leu Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(C)C)C(=O)O)NC(=O)[C@H](CCSC)N WPTDJKDGICUFCP-XUXIUFHCSA-N 0.000 description 1
- LNXGEYIEEUZGGH-JYJNAYRXSA-N Met-Phe-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CCSC)CC1=CC=CC=C1 LNXGEYIEEUZGGH-JYJNAYRXSA-N 0.000 description 1
- KYXDADPHSNFWQX-VEVYYDQMSA-N Met-Thr-Asp Chemical compound CSCC[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@H](C(O)=O)CC(O)=O KYXDADPHSNFWQX-VEVYYDQMSA-N 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- YBAFDPFAUTYYRW-UHFFFAOYSA-N N-L-alpha-glutamyl-L-leucine Natural products CC(C)CC(C(O)=O)NC(=O)C(N)CCC(O)=O YBAFDPFAUTYYRW-UHFFFAOYSA-N 0.000 description 1
- KZNQNBZMBZJQJO-UHFFFAOYSA-N N-glycyl-L-proline Natural products NCC(=O)N1CCCC1C(O)=O KZNQNBZMBZJQJO-UHFFFAOYSA-N 0.000 description 1
- JIYJYFIXQTYDNF-YDHLFZDLSA-N Phe-Asn-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CC=CC=C1)N JIYJYFIXQTYDNF-YDHLFZDLSA-N 0.000 description 1
- ZLGQEBCCANLYRA-RYUDHWBXSA-N Phe-Gly-Glu Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)NCC(=O)N[C@@H](CCC(O)=O)C(O)=O ZLGQEBCCANLYRA-RYUDHWBXSA-N 0.000 description 1
- SFKOEHXABNPLRT-KBPBESRZSA-N Phe-His-Gly Chemical compound N[C@@H](Cc1ccccc1)C(=O)N[C@@H](Cc1cnc[nH]1)C(=O)NCC(O)=O SFKOEHXABNPLRT-KBPBESRZSA-N 0.000 description 1
- SHUFSZDAIPLZLF-BEAPCOKYSA-N Phe-Thr-Pro Chemical compound C[C@H]([C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=CC=C2)N)O SHUFSZDAIPLZLF-BEAPCOKYSA-N 0.000 description 1
- GTMSCDVFQLNEOY-BZSNNMDCSA-N Phe-Tyr-Asn Chemical compound C1=CC=C(C=C1)C[C@@H](C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CC(=O)N)C(=O)O)N GTMSCDVFQLNEOY-BZSNNMDCSA-N 0.000 description 1
- MMPBPRXOFJNCCN-ZEWNOJEFSA-N Phe-Tyr-Ile Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O MMPBPRXOFJNCCN-ZEWNOJEFSA-N 0.000 description 1
- XALFIVXGQUEGKV-JSGCOSHPSA-N Phe-Val-Gly Chemical compound OC(=O)CNC(=O)[C@H](C(C)C)NC(=O)[C@@H](N)CC1=CC=CC=C1 XALFIVXGQUEGKV-JSGCOSHPSA-N 0.000 description 1
- NHDVNAKDACFHPX-GUBZILKMSA-N Pro-Arg-Ala Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(O)=O NHDVNAKDACFHPX-GUBZILKMSA-N 0.000 description 1
- CYQQWUPHIZVCNY-GUBZILKMSA-N Pro-Arg-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(O)=O CYQQWUPHIZVCNY-GUBZILKMSA-N 0.000 description 1
- SMCHPSMKAFIERP-FXQIFTODSA-N Pro-Asn-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@@H]1CCCN1 SMCHPSMKAFIERP-FXQIFTODSA-N 0.000 description 1
- ODPIUQVTULPQEP-CIUDSAMLSA-N Pro-Gln-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@@H]1CCCN1 ODPIUQVTULPQEP-CIUDSAMLSA-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
- GFHOSBYCLACKEK-GUBZILKMSA-N Pro-Pro-Asn Chemical compound [H]N1CCC[C@H]1C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(N)=O)C(O)=O GFHOSBYCLACKEK-GUBZILKMSA-N 0.000 description 1
- GZNYIXWOIUFLGO-ZJDVBMNYSA-N Pro-Thr-Thr Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O GZNYIXWOIUFLGO-ZJDVBMNYSA-N 0.000 description 1
- BVRBCQBUNGAWFP-KKUMJFAQSA-N Pro-Tyr-Gln Chemical compound C1C[C@H](NC1)C(=O)N[C@@H](CC2=CC=C(C=C2)O)C(=O)N[C@@H](CCC(=O)N)C(=O)O BVRBCQBUNGAWFP-KKUMJFAQSA-N 0.000 description 1
- OOZJHTXCLJUODH-QXEWZRGKSA-N Pro-Val-Asp Chemical compound OC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1 OOZJHTXCLJUODH-QXEWZRGKSA-N 0.000 description 1
- UBRXAVQWXOWRSJ-ZLUOBGJFSA-N Ser-Asn-Asp Chemical compound C([C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CO)N)C(=O)N UBRXAVQWXOWRSJ-ZLUOBGJFSA-N 0.000 description 1
- VGNYHOBZJKWRGI-CIUDSAMLSA-N Ser-Asn-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H](N)CO VGNYHOBZJKWRGI-CIUDSAMLSA-N 0.000 description 1
- UGJRQLURDVGULT-LKXGYXEUSA-N Ser-Asn-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O UGJRQLURDVGULT-LKXGYXEUSA-N 0.000 description 1
- OLIJLNWFEQEFDM-SRVKXCTJSA-N Ser-Asp-Phe Chemical compound OC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 OLIJLNWFEQEFDM-SRVKXCTJSA-N 0.000 description 1
- CDVFZMOFNJPUDD-ACZMJKKPSA-N Ser-Gln-Asn Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O CDVFZMOFNJPUDD-ACZMJKKPSA-N 0.000 description 1
- DSGYZICNAMEJOC-AVGNSLFASA-N Ser-Glu-Phe Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DSGYZICNAMEJOC-AVGNSLFASA-N 0.000 description 1
- IOVHBRCQOGWAQH-ZKWXMUAHSA-N Ser-Gly-Ile Chemical compound [H]N[C@@H](CO)C(=O)NCC(=O)N[C@@H]([C@@H](C)CC)C(O)=O IOVHBRCQOGWAQH-ZKWXMUAHSA-N 0.000 description 1
- UIGMAMGZOJVTDN-WHFBIAKZSA-N Ser-Gly-Ser Chemical compound OC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O UIGMAMGZOJVTDN-WHFBIAKZSA-N 0.000 description 1
- ZOPISOXXPQNOCO-SVSWQMSJSA-N Ser-Ile-Thr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)O)NC(=O)[C@H](CO)N ZOPISOXXPQNOCO-SVSWQMSJSA-N 0.000 description 1
- JLPMFVAIQHCBDC-CIUDSAMLSA-N Ser-Lys-Cys Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CS)C(=O)O)NC(=O)[C@H](CO)N JLPMFVAIQHCBDC-CIUDSAMLSA-N 0.000 description 1
- BYCVMHKULKRVPV-GUBZILKMSA-N Ser-Lys-Gln Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O BYCVMHKULKRVPV-GUBZILKMSA-N 0.000 description 1
- PYTKULIABVRXSC-BWBBJGPYSA-N Ser-Ser-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PYTKULIABVRXSC-BWBBJGPYSA-N 0.000 description 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
- 102000012479 Serine Proteases Human genes 0.000 description 1
- 108010022999 Serine Proteases Proteins 0.000 description 1
- 101000629318 Severe acute respiratory syndrome coronavirus 2 Spike glycoprotein Proteins 0.000 description 1
- 206010042434 Sudden death Diseases 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 230000024932 T cell mediated immunity Effects 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- DFTCYYILCSQGIZ-GCJQMDKQSA-N Thr-Ala-Asn Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(N)=O)C(O)=O DFTCYYILCSQGIZ-GCJQMDKQSA-N 0.000 description 1
- UTSWGQNAQRIHAI-UNQGMJICSA-N Thr-Arg-Phe Chemical compound NC(N)=NCCC[C@H](NC(=O)[C@@H](N)[C@H](O)C)C(=O)N[C@H](C(O)=O)CC1=CC=CC=C1 UTSWGQNAQRIHAI-UNQGMJICSA-N 0.000 description 1
- VIBXMCZWVUOZLA-OLHMAJIHSA-N Thr-Asn-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O VIBXMCZWVUOZLA-OLHMAJIHSA-N 0.000 description 1
- NLSNVZAREYQMGR-HJGDQZAQSA-N Thr-Asp-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O NLSNVZAREYQMGR-HJGDQZAQSA-N 0.000 description 1
- OHAJHDJOCKKJLV-LKXGYXEUSA-N Thr-Asp-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O OHAJHDJOCKKJLV-LKXGYXEUSA-N 0.000 description 1
- LHEZGZQRLDBSRR-WDCWCFNPSA-N Thr-Glu-Leu Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O LHEZGZQRLDBSRR-WDCWCFNPSA-N 0.000 description 1
- JRAUIKJSEAKTGD-TUBUOCAGSA-N Thr-Ile-His Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H]([C@@H](C)O)N JRAUIKJSEAKTGD-TUBUOCAGSA-N 0.000 description 1
- VRUFCJZQDACGLH-UVOCVTCTSA-N Thr-Leu-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O VRUFCJZQDACGLH-UVOCVTCTSA-N 0.000 description 1
- MXNAOGFNFNKUPD-JHYOHUSXSA-N Thr-Phe-Thr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)O)C(O)=O MXNAOGFNFNKUPD-JHYOHUSXSA-N 0.000 description 1
- MUAFDCVOHYAFNG-RCWTZXSCSA-N Thr-Pro-Arg Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCCNC(N)=N)C(O)=O MUAFDCVOHYAFNG-RCWTZXSCSA-N 0.000 description 1
- LKJCABTUFGTPPY-HJGDQZAQSA-N Thr-Pro-Gln Chemical compound C[C@@H](O)[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CCC(N)=O)C(O)=O LKJCABTUFGTPPY-HJGDQZAQSA-N 0.000 description 1
- IVDFVBVIVLJJHR-LKXGYXEUSA-N Thr-Ser-Asp Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(O)=O)C(O)=O IVDFVBVIVLJJHR-LKXGYXEUSA-N 0.000 description 1
- 241000243777 Trichinella spiralis Species 0.000 description 1
- KULBQAVOXHQLIY-HSCHXYMDSA-N Trp-Ile-Leu Chemical compound C1=CC=C2C(C[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O)=CNC2=C1 KULBQAVOXHQLIY-HSCHXYMDSA-N 0.000 description 1
- GRSCONMARGNYHA-PMVMPFDFSA-N Trp-Lys-Phe Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O GRSCONMARGNYHA-PMVMPFDFSA-N 0.000 description 1
- YXSSXUIBUJGHJY-SFJXLCSZSA-N Trp-Thr-Phe Chemical compound C([C@H](NC(=O)[C@@H](NC(=O)[C@@H](N)CC=1C2=CC=CC=C2NC=1)[C@H](O)C)C(O)=O)C1=CC=CC=C1 YXSSXUIBUJGHJY-SFJXLCSZSA-N 0.000 description 1
- AKFLVKKWVZMFOT-IHRRRGAJSA-N Tyr-Arg-Asn 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](CC(N)=O)C(O)=O AKFLVKKWVZMFOT-IHRRRGAJSA-N 0.000 description 1
- CKHQKYHIZCRTAP-SOUVJXGZSA-N Tyr-Gln-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC2=CC=C(C=C2)O)N)C(=O)O CKHQKYHIZCRTAP-SOUVJXGZSA-N 0.000 description 1
- ZRPLVTZTKPPSBT-AVGNSLFASA-N Tyr-Glu-Ser Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(O)=O ZRPLVTZTKPPSBT-AVGNSLFASA-N 0.000 description 1
- HIINQLBHPIQYHN-JTQLQIEISA-N Tyr-Gly-Gly Chemical compound OC(=O)CNC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 HIINQLBHPIQYHN-JTQLQIEISA-N 0.000 description 1
- YIKDYZDNRCNFQB-KKUMJFAQSA-N Tyr-His-Asn Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC2=CN=CN2)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O YIKDYZDNRCNFQB-KKUMJFAQSA-N 0.000 description 1
- NXRGXTBPMOGFID-CFMVVWHZSA-N Tyr-Ile-Asn Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O NXRGXTBPMOGFID-CFMVVWHZSA-N 0.000 description 1
- HHFMNAVFGBYSAT-IGISWZIWSA-N Tyr-Ile-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N HHFMNAVFGBYSAT-IGISWZIWSA-N 0.000 description 1
- NZBSVMQZQMEUHI-WZLNRYEVSA-N Tyr-Thr-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N NZBSVMQZQMEUHI-WZLNRYEVSA-N 0.000 description 1
- PWKMJDQXKCENMF-MEYUZBJRSA-N Tyr-Thr-Leu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O PWKMJDQXKCENMF-MEYUZBJRSA-N 0.000 description 1
- KUXCBJFJURINGF-PXDAIIFMSA-N Tyr-Trp-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](CC3=CC=C(C=C3)O)N KUXCBJFJURINGF-PXDAIIFMSA-N 0.000 description 1
- PMDOQZFYGWZSTK-LSJOCFKGSA-N Val-Gly-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)C(C)C PMDOQZFYGWZSTK-LSJOCFKGSA-N 0.000 description 1
- OEVFFOBAXHBXKM-HSHDSVGOSA-N Val-Trp-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CNC2=CC=CC=C21)NC(=O)[C@H](C(C)C)N)O OEVFFOBAXHBXKM-HSHDSVGOSA-N 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 108010087924 alanylproline Proteins 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910021502 aluminium hydroxide Inorganic materials 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000005349 anion exchange Methods 0.000 description 1
- 230000000890 antigenic effect Effects 0.000 description 1
- 108010077245 asparaginyl-proline Proteins 0.000 description 1
- 108010047857 aspartylglycine Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000010241 blood sampling Methods 0.000 description 1
- 239000013622 capto Q Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000009089 cytolysis Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 238000006073 displacement reaction Methods 0.000 description 1
- 238000010828 elution Methods 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- 108091005608 glycosylated proteins Proteins 0.000 description 1
- 102000035122 glycosylated proteins Human genes 0.000 description 1
- 108010089804 glycyl-threonine Proteins 0.000 description 1
- 108010092114 histidylphenylalanine Proteins 0.000 description 1
- 108010018006 histidylserine Proteins 0.000 description 1
- 230000028996 humoral immune response Effects 0.000 description 1
- 230000004727 humoral immunity Effects 0.000 description 1
- 230000008348 humoral response Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000003780 insertion Methods 0.000 description 1
- 230000037431 insertion Effects 0.000 description 1
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 1
- BPHPUYQFMNQIOC-NXRLNHOXSA-N isopropyl beta-D-thiogalactopyranoside Chemical compound CC(C)S[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O BPHPUYQFMNQIOC-NXRLNHOXSA-N 0.000 description 1
- 230000021633 leukocyte mediated immunity Effects 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 229940126578 oral vaccine Drugs 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108010077112 prolyl-proline Proteins 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 238000010591 solubility diagram Methods 0.000 description 1
- 229940031626 subunit vaccine Drugs 0.000 description 1
- 235000011149 sulphuric acid Nutrition 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 229940096911 trichinella spiralis Drugs 0.000 description 1
- 108010017949 tyrosyl-glycyl-glycine Proteins 0.000 description 1
- 230000005924 vaccine-induced immune response Effects 0.000 description 1
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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K39/39—Medicinal preparations containing antigens or antibodies characterised by the immunostimulating additives, e.g. chemical adjuvants
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55505—Inorganic adjuvants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/555—Medicinal preparations containing antigens or antibodies characterised by a specific combination antigen/adjuvant
- A61K2039/55511—Organic adjuvants
- A61K2039/55544—Bacterial toxins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K2319/00—Fusion polypeptide
- C07K2319/55—Fusion polypeptide containing a fusion with a toxin, e.g. diphteria toxin
-
- 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
- C12N2720/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsRNA viruses
- C12N2720/00011—Details
- C12N2720/12011—Reoviridae
- C12N2720/12311—Rotavirus, e.g. rotavirus A
- C12N2720/12322—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
- C12N2720/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA dsRNA viruses
- C12N2720/00011—Details
- C12N2720/12011—Reoviridae
- C12N2720/12311—Rotavirus, e.g. rotavirus A
- C12N2720/12334—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)
- General Health & Medical Sciences (AREA)
- Virology (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Oncology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Communicable Diseases (AREA)
- Gastroenterology & Hepatology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了一种CTB/LTB多肽与VP8的融合蛋白，其氨基酸序列包括人轮状病毒VP8蛋白氨基酸片段和外源多肽片段组成，所述外源多肽片段为CTB‑1、CTB‑2和/或LTB‑1，所述CTB‑1的氨基酸序列如SEQ ID NO:1所示，LTB‑1的氨基酸序列如SEQ ID NO:2所示，CTB‑2的氨基酸序列如SEQ ID NO:6所示，所述VP8蛋白氨基酸片段是人轮状病毒Wa株蛋白的氨基酸片段第64‑223位氨基酸序列，其氨基酸序列如SEQ ID NO:3。本发明从CTB和LTB中筛选出多肽片段CTB‑1、CTB‑2和LTB‑1，将多肽片段与轮状病毒的VP8蛋白进行融合表达，分别纯化得到3个多肽‑VP8融合蛋白，具有较好的蛋白水溶性，并且能产生高免疫原性。
Description
技术领域
本发明涉及分子生物学领域，具体涉及一种CTB/LTB多肽与VP8的融合蛋白及轮状病毒疫苗。
背景技术
轮状病毒引起的腹泻是5岁以下儿童最常见的疾病之一。一个孩子在5岁前的几个月里都患有腹泻，这些症状也与成长缓慢有关（Glass R I ,Lang D R ,Ivanoff B N , etal. Introduction: Rotavirus-From Basic Research ToA Vaccine[J]. Journal ofInfectious Diseases(Supplement_1):S1.）。这些数据引发了儿科医生和公共卫生专家的关注，预防和治疗轮状腹泻疾病成为提高儿童存活率的一个关键目标。
早在1998年，Wyeth推出了口服的轮状病毒疫苗Rotashield，使用一年期间，发现接种的儿童发生肠套叠的风险增高，因此，产品不得不撤市。后来Merck和GSK分别推出了口服轮状病毒活疫苗，这两个产品在全球80多个国家广泛应用，对控制轮状病毒造成的严重脱水性腹泻，以及其引起的住院和儿童死亡起到非常好的保护作用。由于口服轮状病毒活疫苗在低收入国家只有大约60%的保护作用，尽管如此，许多这些国家的疫苗覆盖率仍保持在大约70%（Glass RI, Jiang B, Parashar U. The future control of rotavirusdisease: can live oralvaccines alone solve the rotavirus problem. Vaccine2018; 36:2233–6），仅口服活疫苗无法完全控制轮状病毒疾病，人们未来想控制严重轮状病毒疾病的造成的公共卫生健康问题，新一代疫苗的研究将是非常必要的。
发明内容
为构建高可溶性的重组亚单位疫苗，本发明提供了一种CTB/LTB多肽与VP8的融合蛋白，此融合蛋白具有高免疫原性和较好的可溶性。还公开了一种轮状疫苗，能使得免疫机体产生较高的特异性抗体IgG、IgG1和IgG2a。
本发明通过下述技术方案实现：
一种CTB/LTB多肽与VP8的融合蛋白，其氨基酸序列包括人轮状病毒VP8蛋白氨基酸片段和外源多肽片段组成，所述外源多肽片段为CTB-1，CTB-2和/或LTB-1，所述CTB-1的氨基酸序列如SEQ ID NO:1所示，LTB-1的氨基酸序列如SEQ ID NO:2所示，CTB-2的氨基酸序列如SEQ ID NO:6所示，所述VP8蛋白氨基酸片段是人轮状病毒Wa株蛋白的氨基酸片段第64-223位氨基酸序列，其氨基酸序列如SEQ ID NO:3。
融合蛋白为（1）CTB-VP8 P[8]，其氨基酸序列如SEQ ID NO:4所示；（2）LTB1-VP8 P[8]，其氨基酸序列如SEQ ID NO:5所示。3）CTB2-VP8 P[8]，其氨基酸序列如SEQ ID NO:7所示。
一种轮状病毒疫苗，包含如前所述的融合蛋白。
进一步的，所述融合蛋白为10-100微克。
进一步的，还包含医学上可适用的佐剂。
进一步的，所述融合蛋白和佐剂体积比为1：1。
本发明与现有技术相比，具有如下的优点和有益效果：
本发明从CTB和LTB中筛选出多肽片段CTB-1、CTB-2和LTB-1，将多肽片段与轮状病毒的VP8蛋白进行融合表达，分别纯化得到3个多肽-VP8融合蛋白，具有较好的蛋白水溶性，并且能产生高免疫原性，所制备得到的疫苗能够使得小鼠产生较高的特异性抗体IgG、IgG1和IgG2a。
附图说明
此处所说明的附图用来提供对本发明实施例的进一步理解，构成本申请的一部分，并不构成对本发明实施例的限定。在附图中：
图1为本发明融合蛋白合成图；
图2为融合蛋白水溶性图；
图3为纯化效果图；
图4为免疫特异性结果图。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚明白，下面结合实施例和附图，对本发明作进一步的详细说明，本发明的示意性实施方式及其说明仅用于解释本发明，并不作为对本发明的限定。
实施例1
表达系统的构建。
本发明通过对CTB和LTB蛋白的一级结构进行T细胞抗原表位分析筛选出2个多肽片段（如表一所示），其中一条来自CTB-1（含T细胞抗原表位），一条来自LTB-1（含T细胞抗原表位）。（CTB-1：霍乱肠毒素B亚单位；LTB-1：大肠杆菌不耐热肠毒素B亚单位）。本发明还筛选出了一条不含T细胞抗原表位的多肽片段CTB-2，其氨基酸序列如Seq ID NO：6。
表一外源多肽序列
| 多肽 | 氨基酸序列 |
| CTB-1 | Seq ID NO：1 HGTPQNITDLCAEYHNTQI |
| LTB-1 | Seq ID NO：2 GAPQTITELCSEYRNTQI |
| CTB-2 | Seq ID NO：6 TDLCAEYHNTQIYTLNDKIFS |
在本实施案例中，将CTB-1、CTB-2和LTB-1多肽与轮状病毒的VP8蛋白（AA64-223）进行融合表达，将得到如下融合蛋白（不含外源多肽的VP8P[8]本身作为对照）： 1）VP8P[8]
2）CTB1-VP8P[8]融合蛋白，氨基酸序列如Seq ID NO：4所示；
3）LTB1-VP8P[8] 融合蛋白，氨基酸序列如Seq ID NO：5所示；
4）CTB2-VP8P[8] 融合蛋白，氨基酸序列如Seq ID NO：7所示；
表二轮状病毒VP8蛋白和融合蛋白的氨基酸序列
| 轮状病毒毒株 | VP4 P型 | VP8蛋白氨基酸序列 |
| Wa株（GenBank:MT025868） | VP4 P[8] | Seq ID NO：3 |
其中VP8P[8]的序列来源于Rotavirus Astrain Wa isolate RVA/Human-lab/USA/G1P8/Wa/virulent VP4 (VP4) gene（GenBank:MT025868）的蛋白序列的氨基酸集团64-223。为了防止外源多肽影响VP8蛋白的2级和3级结构，在外源多肽和VP8之间增加了一个链接肽（GSGSG），链接肽由不带正电或负电荷的氨基酸组成，所以不影响蛋白的等电点。
融合蛋白的DNA序列交给北京擎科生物技术有限公司进行全序列合成，密码子按照大肠杆菌表达系统进行优化，优化后的序列：
Seq ID NO 8：霍乱肠毒素B亚单位（CTB1）多肽核酸序列；
Seq ID NO 9：霍乱肠毒素B亚单位（CTB2）多肽核酸序列；
Seq ID NO 10：大肠杆菌不耐热肠毒素B亚单位（LTB）多肽核酸序列；
Seq ID NO 11：轮状病毒VP8 P[8]蛋白核酸序列（AA64-223）；
Seq ID NO 12： CTB1-VP8核酸序列；
Seq ID NO 13： CTB2-VP8核酸序列；
Seq ID NO 14： LTB-VP8核酸序列；
Seq ID NO 8：霍乱肠毒素B亚单位（CTB1）多肽核酸序列
CATGGCACCCCGCAGAATATTACCGATCTGTGCGCCGAATATCATAATACCCAGATT；
Seq ID NO 9：霍乱肠毒素B亚单位（CTB2）多肽核酸序列
ACCGATCTGTGTGCCGAATATCATAATACCCAGATCTATACCCTGAATGATAAAATTTTC；
Seq ID NO 10：大肠杆菌不耐热肠毒素B亚单位（LTB）多肽核酸序列
GGTGCACCTCAGACCATTACCGAACTGTGCAGTGAATATCGTAATACCCAGATT；
Seq ID NO 11：轮状病毒VP8 P[8]蛋白核酸序列（AA64-223）
ATGATTCTGGATGGTCCGTATCAGCCGACCACCTTTACCCCGCCGAATGATTATTGGATTCTGATTAATAGTAACACCAACGGCGTTGTGTATGAAAGCACCAATAATAGCGATTTTTGGACCGCCGTTGTGGCCATTGAACCGCATGTGAATCCGGTTGATCGCCAGTATACCATTTTTGGCGAAAGCAAACAGTTTAATGTGAGCAATGATAGTAACAAATGGAAATTTCTGGAGATGTTTCGCAGCAGCAGTCAGAATGAATTTTATAATCGTCGTACCCTGACCAGTGATACCCGTTTTGTGGGCATTCTGAAATATGGCGGCCGCGTTTGGACCTTTCATGGCGAAACCCCGCGTGCAACCACCGATAGCAGTAGTACCGCAAATCTGAATAATATTAGCATTACCATCCACAGCGAATTTTATATTATCCCGCGCAGTCAGGAAAGTAAATGTAATGAATATATCAACAACGGCCTG；
Seq ID NO 12：CTB1-VP8核酸序列
ATGCATGGCACCCCGCAGAATATTACCGATCTGTGCGCCGAATATCATAATACCCAGATTGGTAGTGGTAGCGGCATTCTGGATGGCCCGTATCAGCCGACCACCTTTACCCCGCCGAATGATTATTGGATTCTGATTAATAGCAACACCAATGGCGTTGTTTATGAAAGCACCAATAATAGCGATTTTTGGACCGCAGTTGTTGCCATTGAACCGCATGTGAATCCGGTTGATCGCCAGTATACCATTTTTGGCGAAAGCAAACAGTTTAATGTGAGTAATGATAGCAACAAATGGAAATTTCTGGAAATGTTTCGCAGTAGTAGTCAGAATGAATTTTATAATCGCCGTACCCTGACCAGTGATACCCGTTTTGTTGGCATTCTGAAATATGGCGGTCGCGTTTGGACCTTTCATGGTGAAACCCCGCGTGCAACCACCGATAGTAGTAGTACCGCAAATCTGAATAATATTAGTATCACCATCCACAGCGAATTTTATATTATTCCGCGTAGTCAGGAAAGCAAATGCAATGAATATATTAACAACGGTCTG
Seq ID NO 13：CTB2-VP8核酸序列
ATGACCGATCTGTGTGCCGAATATCATAATACCCAGATCTATACCCTGAATGATAAAATTTTCGGTAGTGGTAGTGGTATTCTGGATGGTCCGTATCAGCCGACCACCTTTACCCCGCCGAATGATTATTGGATTCTGATTAATAGTAACACCAACGGCGTGGTTTATGAAAGCACCAATAATAGTGATTTCTGGACCGCAGTTGTTGCAATTGAACCGCATGTTAATCCGGTTGATCGCCAGTATACCATTTTTGGCGAAAGCAAACAGTTTAATGTGAGCAATGATAGCAATAAGTGGAAATTTCTGGAAATGTTTCGCAGCAGCAGCCAGAATGAATTTTATAATCGTCGCACCCTGACCAGCGATACCCGTTTTGTTGGTATTCTGAAATATGGCGGCCGCGTTTGGACCTTTCATGGTGAAACCCCGCGTGCCACCACCGATAGTAGCAGCACCGCAAATCTGAATAATATTAGTATTACCATCCACAGTGAGTTTTATATTATCCCGCGCAGCCAGGAAAGCAAATGTAATGAATATATTAACAACGGCCTG；
Seq ID NO 14：LTB1-VP8核酸序列
ATGGGTGCACCTCAGACCATTACCGAACTGTGCAGTGAATATCGTAATACCCAGATTGGCAGTGGCAGCGGTATTCTGGATGGTCCGTATCAGCCGACCACCTTTACCCCGCCGAATGATTATTGGATTCTGATTAATAGTAACACCAACGGCGTTGTGTATGAAAGCACCAATAATAGCGATTTTTGGACCGCCGTTGTGGCCATTGAACCGCATGTGAATCCGGTTGATCGCCAGTATACCATTTTTGGCGAAAGCAAACAGTTTAATGTGAGCAATGATAGTAACAAATGGAAATTTCTGGAGATGTTTCGCAGCAGCAGTCAGAATGAATTTTATAATCGTCGTACCCTGACCAGTGATACCCGTTTTGTGGGCATTCTGAAATATGGCGGCCGCGTTTGGACCTTTCATGGCGAAACCCCGCGTGCAACCACCGATAGCAGTAGTACCGCAAATCTGAATAATATTAGCATTACCATCCACAGCGAATTTTATATTATCCCGCGCAGTCAGGAAAGTAAATGTAATGAATATATCAACAACGGCCTG。
将合成好的目的序列构建到pET30a的NdeI和XhoI酶切位点之间（质粒构建过程由北京擎科生物科技有限公司完成），如图1所示。然后将有目标基因的质粒转化入BL21(DE3)感受态菌体内，进行抗生素筛选。以T7启动子引物为测序引物，通过检测（北京擎科生物科技有限公司）证实目的片段插入PET30a载体的NdeI和XhoI酶切位点之间（Seq ID NO 12、Seq ID NO 13和Seq ID NO 14）。
实施例2
菌种培养和诱导表达。
VP8蛋白是一个非糖基化的蛋白，因此，选择了大肠杆菌表达系统。所用的质粒为pET30a，宿主细胞E.coli BL21(DE3)。挑取生长好的菌落在摇瓶中37 ℃，220rpm培养至OD值约为0.5-0.8，然后把培养的温度降低到16 ℃，加入终浓度为0.2 mM的IPTG诱导表达6-8个小时，诱导结束后将菌体收集，收集条件为4℃，4000 rpm离心30分钟。通过计算，1L的培养体积能够得到5-8g（湿重）的菌。SDS-PAGE（浓缩胶12.5%）检测分析证明菌体表达的蛋白都是可溶性蛋白，SDS-PAGE的结果显示目的蛋白大小与理论值相近，如图2所示。
实施例3
蛋白纯化。
将收集的菌体（LTB1-VP8 /CTB1-VP8/ CTB2-VP8）用buffer A（10mM PH 8.0Tris-HCl+500mM NaCl+30mM 咪唑）重悬，高压裂解后离心取上清。用buffer A平衡Ni-NTANUPharose FF层析柱子（纽龙，Ni-NTA NUPharose FF，5ml），流速为1mL/min。将样品以1mL/min通过亲和柱，上样结束之后，用buffer A用冲洗亲和柱10个柱体积。蛋白的洗脱条件为：60%的buffer B（10mM PH 8.0Tris-HCl+500mM 咪唑）将目的蛋白洗脱下来。
将蛋白通过10kDa MWCO的离心式过滤器（Millipore）进行缓冲液的置换，将缓冲液置换成Q buffer A（10mM pH 8.0 Tris-HCl）。将置换缓冲液之后的样品以1mL/min的流速通过已经事先平衡好的阴离子交换Q柱（Cytiva，Capto Q，5mL）；目的蛋白洗脱条件是通过0-100% Qbuffer B（10mM pH 8.0 Tris-HCl+0.5 M NaCl）线性梯度进行洗脱10个柱体积。纯化之后的目的蛋白通过SDS-PAGE检测蛋白的纯化效果，如图3所示。
实施例4
抗原蛋白免疫原性研究。
疫苗的配制。在本实施例中，在疫苗的配置中加入了氢氧化铝佐剂（Invivogen,vac-alu-250）。配制的方法是用100微升的蛋白（分别含10微克、50微克和100微克蛋白）与100微升的氢氧化铝佐剂1：1混合乳化，然后放于4℃冰箱，准备免疫。
动物实验的分组（本实验的动物小鼠均为健康的BALB/c小鼠（Mus musculus）（雄性），6-8周龄，22.5 g ± 2.5 g。），本试验中，小鼠采用随机区组法分成11组，每组6只，将小鼠分为对照组和实验组，其中对照组包含阴性对照组（0.01 M PBS，标号为：PBS组）和佐剂对照组（铝佐剂），分别选择融合蛋白CTB1-VP8组，CTB2-VP8组，LTB1-VP8 组和VP8组，不同的蛋白免疫剂量即10 μg、50 μg和100 μg免疫实验组。
疫苗的免疫接种：免疫途径为后腿肌肉注射，免疫三次，一次基础免疫，两次加强免疫，每次免疫间隔两周。实验组中，每只小鼠每次免疫接种疫苗为200 μL（含100 μL融合蛋白稀释液和100 μL佐剂）。以相同途径免疫阴性对照PBS组（每只小鼠每次免疫接种200 μL的0.01 M PBS）和佐剂对照AL组（每只小鼠每次免疫接种100 μL的0.01 M PBS和100 μL佐剂）。
免疫接种后，观察并记录各组小鼠的情况，主要观察小鼠有无突发性死亡等情况。采血方式：采用断尾取血的方式进行收集小鼠血液，最后一次采血后可以直接将其处死。（注意：每只小鼠分别取样）
血液样品的处理方式：
（1）首先收集小鼠的尾部血液，盛于1.5 mL EP管内，至少满足每只每次得到的血清不低于100 µL。
（2）收集血液后37 ℃放置1 h，取出放置4 ℃过夜，然后在4 ℃环境中，5000 rpm离心15 min。
（3）仔细收集析出的上层血清，最后将血清分装（每只每次采血分装2管即可，每管50 µL）做好标记、置于-80 ℃保存。
实施例5
间接ELISA法检测蛋白特异性抗体IgG\IgG1\IgG2a。
取保存于-80℃冰箱的免疫后血清，间接ELISA法检测血清中各蛋白特异性抗体IgG、IgG1、gG2a。具体操作步骤如下：
（1）、包被：用包被液稀释蛋白（抗原），稀释比例为1：200，每孔100 μL，4℃过夜。
（2）、弃蛋白液，拍干，PBST洗3次，每次5 min（尽量加满，但不串孔）。
（3）、封闭：5%脱脂奶粉封闭（加满），37℃，1h30min。
（4）、PBST洗3次，每次5 min。
（5）、待检血清样品稀释：选取阳性血清和阴性血清，用PBS稀释，稀释比例为1：400，每孔100 μL，37℃，1 h。
（6）、PBST洗3次，每次5 min。
（7）、加入相应的IgG、IgG1、gG2a的HRP二抗（按照说明书稀释），每孔100 μL，37℃，1 h。
（8）、PBST洗5次，每次5 min。
（9）、显色：每孔加入TMB，每孔100 μL，37℃，避光15-20 min。
（10）、终止：加入2M H2SO4，每孔100 μL，测OD值（450nm）。
注：IgG二抗（辣根过氧化物酶标记山羊抗小鼠IgG(H+L)，A0216，上海碧云天生物技术有限公司）、IgG1二抗（山羊抗小鼠IgG1 (HRP)，ab97240，艾博抗（上海）贸易有限公司）、IgG2a二抗（山羊抗小鼠IgG2a heavy chain，ab97245，艾博抗（上海）贸易有限公司）。
Th1细胞分泌 IFN-γ，介导细胞免疫反应，Th2细胞分泌IL-4，促进体液免疫反应。但IFN-γ和IL-4水平通常是反向调节的。IL-4诱导Th0细胞分化为Th2细胞，Th2细胞分泌的IL-4促进Th2细胞增殖，引起体液免疫，抑制Th1增殖(Chen, L.et al.IL-4 inducesdifferentiation and expansion of Th2 cytokine-producing eosinophils. 2004,J Immunol172, 2059-2066)，同时该因子能促进抗体向lgG1转换。IFN-γ及IL-12促进Th0向Th1分化，Th1细胞分泌IFN-γ，该细胞因子能促进抗体向lgG2a转换(Prompetchara, E.et al.DNA vaccine candidate encoding SARS-CoV-2 spike proteins elicited potenthumoral and Th1 cell-mediated immune responses in mice.PLoS One,2021,16,e0248007; Ding, Y.et al.A novel combined vaccine against classical swinefever and porcine epidemic diarrhea viruseselicits a significant Th2-favoredhumoral response in mice.Vaccine,2021,39, 4573-4576)。在小鼠体内，Th1型会产生lgG2a型抗体，Th2型会产生lgG1型抗体，通常使用lgG1/lgG2a来评价疫苗诱导的免疫应答中占主导地位的免疫应答(Yue, X.et al.Molecular characterization of aTrichinella spiralis serine proteinase.Vet Res,2020,51, 125)。
结果表3和如图4所示，在特异性抗体指标和IgG1指标中可以看出：CTB1-VP8和LTB1-VP8蛋白的免疫效果显著高于CTB2-VP8蛋白组、VP8蛋白组、PBS对照组和AL佐剂对照组（p<0.001），然而CTB2-VP8蛋白和VP8蛋白免疫的效果与PBS对照组和AL佐剂对照组没有明显的差异（p>0.001），其中CTB1-VP8蛋白免疫组的效果最好。在IgG2a指标中可以看出：CTB1-VP8和LTB1-VP8蛋白的免疫效果于VP8蛋白免疫效果相比，CTB1-VP8和LTB1-VP8蛋白免疫效果在第二次免疫呈现上升的趋势，并在42 Day时与CTB2-VP8蛋白组、VP8蛋白组、PBS对照组和AL佐剂对照组有明显的差异（p<0.05），在特异性抗体指标、IgG1指标和IgG2a指标中，VP8蛋白组与PBS对照组和AL佐剂对照组没有明显的差异（p>0.05）。总的来说：CTB1-VP8和LTB1-VP8能刺激小鼠产生较高的特异性抗体和IgG1和IgG2a，能引起小鼠体内产生较好的免疫应答。
表3 动物免疫后血清中特异性抗体、IgG1和IgG2a数据
本发明涉及到的氨基酸序列表如下：
Seq ID NO 1：霍乱肠毒素B亚单位（CTB1）多肽氨基酸序列；
Seq ID NO 2：大肠杆菌不耐热肠毒素B亚单位（LTB）多肽氨基酸序列；
Seq ID NO 3：轮状病毒VP8 P[8]蛋白氨基酸序列（AA64-223）；
Seq ID NO 4：CTB1-VP8氨基酸序列；
Seq ID NO 5：LTB-VP8氨基酸序列；
Seq ID NO 6：霍乱肠毒素B亚单位（CTB2）多肽氨基酸序列；
Seq ID NO 7：CTB2-VP8氨基酸序列；
Seq ID NO 8：霍乱肠毒素B亚单位（CTB1）多肽核酸序列；
Seq ID NO 9：霍乱肠毒素B亚单位（CTB2）多肽核酸序列；
Seq ID NO 10：大肠杆菌不耐热肠毒素B亚单位（LTB）多肽核酸序列；
Seq ID NO 11：轮状病毒VP8 P[8]蛋白核酸序列（AA64-223）；
Seq ID NO 12：CTB1-VP8核酸序列；
Seq ID NO 13：CTB2-VP8核酸序列；
Seq ID NO 14：LTB1-VP8核酸序列；
Seq ID NO 1：HGTPQNITDLCAEYHNTQI；
Seq ID NO 2：GAPQTITELCSEYRNTQI；
Seq ID NO 3：
MILDGPYQPTTFTPPNDYWILINSNTNGVVYESTNNSDFWTAVVAIEPHVNPVDRQYTIFGESKQFNVSNDSNKWKFLEMFRSSSQNEFYNRRTLTSDTRFVGILKYGGRVWTFHGETPRATTDSSSTANLNNISITIHSEFYIIPRSQESKCNEYINNGL；
Seq ID NO 4：
MHGTPQNITDLCAEYHNTQIGSGSGILDGPYQPTTFTPPNDYWILINSNTNGVVYESTNNSDFWTAVVAIEPHVNPVDRQYTIFGESKQFNVSNDSNKWKFLEMFRSSSQNEFYNRRTLTSDTRFVGILKYGGRVWTFHGETPRATTDSSSTANLNNISITIHSEFYIIPRSQESKCNEYINNGL；
Seq ID NO 5：
MGAPQTITELCSEYRNTQIGSGSGILDGPYQPTTFTPPNDYWILINSNTNGVVYESTNNSDFWTAVVAIEPHVNPVDRQYTIFGESKQFNVSNDSNKWKFLEMFRSSSQNEFYNRRTLTSDTRFVGILKYGGRVWTFHGETPRATTDSSSTANLNNISITIHSEFYIIPRSQESKCNEYINNGL；
Seq ID NO 6：TDLCAEYHNTQIYTLNDKIF；
Seq ID NO 7：
MTDLCAEYHNTQIYTLNDKIFGSGSGILDGPYQPTTFTPPNDYWILINSNTNGVVYESTNNSDFWTAVVAIEPHVNPVDRQYTIFGESKQFNVSNDSNKWKFLEMFRSSSQNEFYNRRTLTSDTRFVGILKYGGRVWTFHGETPRATTDSSSTANLNNISITIHSEFYIIPRSQESKCNEYINNGL；
Seq ID NO 8：霍乱肠毒素B亚单位（CTB1）多肽核酸序列
CATGGCACCCCGCAGAATATTACCGATCTGTGCGCCGAATATCATAATACCCAGATT；
Seq ID NO 9：霍乱肠毒素B亚单位（CTB2）多肽核酸序列
ACCGATCTGTGTGCCGAATATCATAATACCCAGATCTATACCCTGAATGATAAAATTTTC；
Seq ID NO 10：大肠杆菌不耐热肠毒素B亚单位（LTB）多肽核酸序列
GGTGCACCTCAGACCATTACCGAACTGTGCAGTGAATATCGTAATACCCAGATT；
Seq ID NO 11：轮状病毒VP8 P[8]蛋白核酸序列（AA64-223）
ATGATTCTGGATGGTCCGTATCAGCCGACCACCTTTACCCCGCCGAATGATTATTGGATTCTGATTAATAGTAACACCAACGGCGTTGTGTATGAAAGCACCAATAATAGCGATTTTTGGACCGCCGTTGTGGCCATTGAACCGCATGTGAATCCGGTTGATCGCCAGTATACCATTTTTGGCGAAAGCAAACAGTTTAATGTGAGCAATGATAGTAACAAATGGAAATTTCTGGAGATGTTTCGCAGCAGCAGTCAGAATGAATTTTATAATCGTCGTACCCTGACCAGTGATACCCGTTTTGTGGGCATTCTGAAATATGGCGGCCGCGTTTGGACCTTTCATGGCGAAACCCCGCGTGCAACCACCGATAGCAGTAGTACCGCAAATCTGAATAATATTAGCATTACCATCCACAGCGAATTTTATATTATCCCGCGCAGTCAGGAAAGTAAATGTAATGAATATATCAACAACGGCCTG；
Seq ID NO 12：CTB1-VP8核酸序列
ATGCATGGCACCCCGCAGAATATTACCGATCTGTGCGCCGAATATCATAATACCCAGATTGGTAGTGGTAGCGGCATTCTGGATGGCCCGTATCAGCCGACCACCTTTACCCCGCCGAATGATTATTGGATTCTGATTAATAGCAACACCAATGGCGTTGTTTATGAAAGCACCAATAATAGCGATTTTTGGACCGCAGTTGTTGCCATTGAACCGCATGTGAATCCGGTTGATCGCCAGTATACCATTTTTGGCGAAAGCAAACAGTTTAATGTGAGTAATGATAGCAACAAATGGAAATTTCTGGAAATGTTTCGCAGTAGTAGTCAGAATGAATTTTATAATCGCCGTACCCTGACCAGTGATACCCGTTTTGTTGGCATTCTGAAATATGGCGGTCGCGTTTGGACCTTTCATGGTGAAACCCCGCGTGCAACCACCGATAGTAGTAGTACCGCAAATCTGAATAATATTAGTATCACCATCCACAGCGAATTTTATATTATTCCGCGTAGTCAGGAAAGCAAATGCAATGAATATATTAACAACGGTCTG
Seq ID NO 13：CTB2-VP8核酸序列
ATGACCGATCTGTGTGCCGAATATCATAATACCCAGATCTATACCCTGAATGATAAAATTTTCGGTAGTGGTAGTGGTATTCTGGATGGTCCGTATCAGCCGACCACCTTTACCCCGCCGAATGATTATTGGATTCTGATTAATAGTAACACCAACGGCGTGGTTTATGAAAGCACCAATAATAGTGATTTCTGGACCGCAGTTGTTGCAATTGAACCGCATGTTAATCCGGTTGATCGCCAGTATACCATTTTTGGCGAAAGCAAACAGTTTAATGTGAGCAATGATAGCAATAAGTGGAAATTTCTGGAAATGTTTCGCAGCAGCAGCCAGAATGAATTTTATAATCGTCGCACCCTGACCAGCGATACCCGTTTTGTTGGTATTCTGAAATATGGCGGCCGCGTTTGGACCTTTCATGGTGAAACCCCGCGTGCCACCACCGATAGTAGCAGCACCGCAAATCTGAATAATATTAGTATTACCATCCACAGTGAGTTTTATATTATCCCGCGCAGCCAGGAAAGCAAATGTAATGAATATATTAACAACGGCCTG；
Seq ID NO 14：LTB1-VP8核酸序列
ATGGGTGCACCTCAGACCATTACCGAACTGTGCAGTGAATATCGTAATACCCAGATTGGCAGTGGCAGCGGTATTCTGGATGGTCCGTATCAGCCGACCACCTTTACCCCGCCGAATGATTATTGGATTCTGATTAATAGTAACACCAACGGCGTTGTGTATGAAAGCACCAATAATAGCGATTTTTGGACCGCCGTTGTGGCCATTGAACCGCATGTGAATCCGGTTGATCGCCAGTATACCATTTTTGGCGAAAGCAAACAGTTTAATGTGAGCAATGATAGTAACAAATGGAAATTTCTGGAGATGTTTCGCAGCAGCAGTCAGAATGAATTTTATAATCGTCGTACCCTGACCAGTGATACCCGTTTTGTGGGCATTCTGAAATATGGCGGCCGCGTTTGGACCTTTCATGGCGAAACCCCGCGTGCAACCACCGATAGCAGTAGTACCGCAAATCTGAATAATATTAGCATTACCATCCACAGCGAATTTTATATTATCCCGCGCAGTCAGGAAAGTAAATGTAATGAATATATCAACAACGGCCTG。
以上所述的具体实施方式，对本发明的目的、技术方案和有益效果进行了进一步详细说明，所应理解的是，以上所述仅为本发明的具体实施方式而已，并不用于限定本发明的保护范围，凡在本发明的精神和原则之内，所做的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。
序列表
<110> 成都康华生物制品股份有限公司
<120> 一种CTB/LTB多肽与VP8的融合蛋白及轮状病毒疫苗
<130> 2022.04.06
<160> 14
<170> SIPOSequenceListing 1.0
<210> 1
<211> 19
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 1
His Gly Thr Pro Gln Asn Ile Thr Asp Leu Cys Ala Glu Tyr His Asn
1 5 10 15
Thr Gln Ile
<210> 2
<211> 18
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 2
Gly Ala Pro Gln Thr Ile Thr Glu Leu Cys Ser Glu Tyr Arg Asn Thr
1 5 10 15
Gln Ile
<210> 3
<211> 161
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 3
Met Ile Leu Asp Gly Pro Tyr Gln Pro Thr Thr Phe Thr Pro Pro Asn
1 5 10 15
Asp Tyr Trp Ile Leu Ile Asn Ser Asn Thr Asn Gly Val Val Tyr Glu
20 25 30
Ser Thr Asn Asn Ser Asp Phe Trp Thr Ala Val Val Ala Ile Glu Pro
35 40 45
His Val Asn Pro Val Asp Arg Gln Tyr Thr Ile Phe Gly Glu Ser Lys
50 55 60
Gln Phe Asn Val Ser Asn Asp Ser Asn Lys Trp Lys Phe Leu Glu Met
65 70 75 80
Phe Arg Ser Ser Ser Gln Asn Glu Phe Tyr Asn Arg Arg Thr Leu Thr
85 90 95
Ser Asp Thr Arg Phe Val Gly Ile Leu Lys Tyr Gly Gly Arg Val Trp
100 105 110
Thr Phe His Gly Glu Thr Pro Arg Ala Thr Thr Asp Ser Ser Ser Thr
115 120 125
Ala Asn Leu Asn Asn Ile Ser Ile Thr Ile His Ser Glu Phe Tyr Ile
130 135 140
Ile Pro Arg Ser Gln Glu Ser Lys Cys Asn Glu Tyr Ile Asn Asn Gly
145 150 155 160
Leu
<210> 4
<211> 185
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 4
Met His Gly Thr Pro Gln Asn Ile Thr Asp Leu Cys Ala Glu Tyr His
1 5 10 15
Asn Thr Gln Ile Gly Ser Gly Ser Gly Ile Leu Asp Gly Pro Tyr Gln
20 25 30
Pro Thr Thr Phe Thr Pro Pro Asn Asp Tyr Trp Ile Leu Ile Asn Ser
35 40 45
Asn Thr Asn Gly Val Val Tyr Glu Ser Thr Asn Asn Ser Asp Phe Trp
50 55 60
Thr Ala Val Val Ala Ile Glu Pro His Val Asn Pro Val Asp Arg Gln
65 70 75 80
Tyr Thr Ile Phe Gly Glu Ser Lys Gln Phe Asn Val Ser Asn Asp Ser
85 90 95
Asn Lys Trp Lys Phe Leu Glu Met Phe Arg Ser Ser Ser Gln Asn Glu
100 105 110
Phe Tyr Asn Arg Arg Thr Leu Thr Ser Asp Thr Arg Phe Val Gly Ile
115 120 125
Leu Lys Tyr Gly Gly Arg Val Trp Thr Phe His Gly Glu Thr Pro Arg
130 135 140
Ala Thr Thr Asp Ser Ser Ser Thr Ala Asn Leu Asn Asn Ile Ser Ile
145 150 155 160
Thr Ile His Ser Glu Phe Tyr Ile Ile Pro Arg Ser Gln Glu Ser Lys
165 170 175
Cys Asn Glu Tyr Ile Asn Asn Gly Leu
180 185
<210> 5
<211> 184
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 5
Met Gly Ala Pro Gln Thr Ile Thr Glu Leu Cys Ser Glu Tyr Arg Asn
1 5 10 15
Thr Gln Ile Gly Ser Gly Ser Gly Ile Leu Asp Gly Pro Tyr Gln Pro
20 25 30
Thr Thr Phe Thr Pro Pro Asn Asp Tyr Trp Ile Leu Ile Asn Ser Asn
35 40 45
Thr Asn Gly Val Val Tyr Glu Ser Thr Asn Asn Ser Asp Phe Trp Thr
50 55 60
Ala Val Val Ala Ile Glu Pro His Val Asn Pro Val Asp Arg Gln Tyr
65 70 75 80
Thr Ile Phe Gly Glu Ser Lys Gln Phe Asn Val Ser Asn Asp Ser Asn
85 90 95
Lys Trp Lys Phe Leu Glu Met Phe Arg Ser Ser Ser Gln Asn Glu Phe
100 105 110
Tyr Asn Arg Arg Thr Leu Thr Ser Asp Thr Arg Phe Val Gly Ile Leu
115 120 125
Lys Tyr Gly Gly Arg Val Trp Thr Phe His Gly Glu Thr Pro Arg Ala
130 135 140
Thr Thr Asp Ser Ser Ser Thr Ala Asn Leu Asn Asn Ile Ser Ile Thr
145 150 155 160
Ile His Ser Glu Phe Tyr Ile Ile Pro Arg Ser Gln Glu Ser Lys Cys
165 170 175
Asn Glu Tyr Ile Asn Asn Gly Leu
180
<210> 6
<211> 20
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 6
Thr Asp Leu Cys Ala Glu Tyr His Asn Thr Gln Ile Tyr Thr Leu Asn
1 5 10 15
Asp Lys Ile Phe
20
<210> 7
<211> 186
<212> PRT
<213> 人工序列(Artificial Sequence)
<400> 7
Met Thr Asp Leu Cys Ala Glu Tyr His Asn Thr Gln Ile Tyr Thr Leu
1 5 10 15
Asn Asp Lys Ile Phe Gly Ser Gly Ser Gly Ile Leu Asp Gly Pro Tyr
20 25 30
Gln Pro Thr Thr Phe Thr Pro Pro Asn Asp Tyr Trp Ile Leu Ile Asn
35 40 45
Ser Asn Thr Asn Gly Val Val Tyr Glu Ser Thr Asn Asn Ser Asp Phe
50 55 60
Trp Thr Ala Val Val Ala Ile Glu Pro His Val Asn Pro Val Asp Arg
65 70 75 80
Gln Tyr Thr Ile Phe Gly Glu Ser Lys Gln Phe Asn Val Ser Asn Asp
85 90 95
Ser Asn Lys Trp Lys Phe Leu Glu Met Phe Arg Ser Ser Ser Gln Asn
100 105 110
Glu Phe Tyr Asn Arg Arg Thr Leu Thr Ser Asp Thr Arg Phe Val Gly
115 120 125
Ile Leu Lys Tyr Gly Gly Arg Val Trp Thr Phe His Gly Glu Thr Pro
130 135 140
Arg Ala Thr Thr Asp Ser Ser Ser Thr Ala Asn Leu Asn Asn Ile Ser
145 150 155 160
Ile Thr Ile His Ser Glu Phe Tyr Ile Ile Pro Arg Ser Gln Glu Ser
165 170 175
Lys Cys Asn Glu Tyr Ile Asn Asn Gly Leu
180 185
<210> 8
<211> 57
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 8
catggcaccc cgcagaatat taccgatctg tgcgccgaat atcataatac ccagatt 57
<210> 9
<211> 60
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 9
accgatctgt gtgccgaata tcataatacc cagatctata ccctgaatga taaaattttc 60
<210> 10
<211> 54
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 10
ggtgcacctc agaccattac cgaactgtgc agtgaatatc gtaataccca gatt 54
<210> 11
<211> 483
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 11
atgattctgg atggtccgta tcagccgacc acctttaccc cgccgaatga ttattggatt 60
ctgattaata gtaacaccaa cggcgttgtg tatgaaagca ccaataatag cgatttttgg 120
accgccgttg tggccattga accgcatgtg aatccggttg atcgccagta taccattttt 180
ggcgaaagca aacagtttaa tgtgagcaat gatagtaaca aatggaaatt tctggagatg 240
tttcgcagca gcagtcagaa tgaattttat aatcgtcgta ccctgaccag tgatacccgt 300
tttgtgggca ttctgaaata tggcggccgc gtttggacct ttcatggcga aaccccgcgt 360
gcaaccaccg atagcagtag taccgcaaat ctgaataata ttagcattac catccacagc 420
gaattttata ttatcccgcg cagtcaggaa agtaaatgta atgaatatat caacaacggc 480
ctg 483
<210> 12
<211> 555
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 12
atgcatggca ccccgcagaa tattaccgat ctgtgcgccg aatatcataa tacccagatt 60
ggtagtggta gcggcattct ggatggcccg tatcagccga ccacctttac cccgccgaat 120
gattattgga ttctgattaa tagcaacacc aatggcgttg tttatgaaag caccaataat 180
agcgattttt ggaccgcagt tgttgccatt gaaccgcatg tgaatccggt tgatcgccag 240
tataccattt ttggcgaaag caaacagttt aatgtgagta atgatagcaa caaatggaaa 300
tttctggaaa tgtttcgcag tagtagtcag aatgaatttt ataatcgccg taccctgacc 360
agtgataccc gttttgttgg cattctgaaa tatggcggtc gcgtttggac ctttcatggt 420
gaaaccccgc gtgcaaccac cgatagtagt agtaccgcaa atctgaataa tattagtatc 480
accatccaca gcgaatttta tattattccg cgtagtcagg aaagcaaatg caatgaatat 540
attaacaacg gtctg 555
<210> 13
<211> 558
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 13
atgaccgatc tgtgtgccga atatcataat acccagatct ataccctgaa tgataaaatt 60
ttcggtagtg gtagtggtat tctggatggt ccgtatcagc cgaccacctt taccccgccg 120
aatgattatt ggattctgat taatagtaac accaacggcg tggtttatga aagcaccaat 180
aatagtgatt tctggaccgc agttgttgca attgaaccgc atgttaatcc ggttgatcgc 240
cagtatacca tttttggcga aagcaaacag tttaatgtga gcaatgatag caataagtgg 300
aaatttctgg aaatgtttcg cagcagcagc cagaatgaat tttataatcg tcgcaccctg 360
accagcgata cccgttttgt tggtattctg aaatatggcg gccgcgtttg gacctttcat 420
ggtgaaaccc cgcgtgccac caccgatagt agcagcaccg caaatctgaa taatattagt 480
attaccatcc acagtgagtt ttatattatc ccgcgcagcc aggaaagcaa atgtaatgaa 540
tatattaaca acggcctg 558
<210> 14
<211> 552
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 14
atgggtgcac ctcagaccat taccgaactg tgcagtgaat atcgtaatac ccagattggc 60
agtggcagcg gtattctgga tggtccgtat cagccgacca cctttacccc gccgaatgat 120
tattggattc tgattaatag taacaccaac ggcgttgtgt atgaaagcac caataatagc 180
gatttttgga ccgccgttgt ggccattgaa ccgcatgtga atccggttga tcgccagtat 240
accatttttg gcgaaagcaa acagtttaat gtgagcaatg atagtaacaa atggaaattt 300
ctggagatgt ttcgcagcag cagtcagaat gaattttata atcgtcgtac cctgaccagt 360
gatacccgtt ttgtgggcat tctgaaatat ggcggccgcg tttggacctt tcatggcgaa 420
accccgcgtg caaccaccga tagcagtagt accgcaaatc tgaataatat tagcattacc 480
atccacagcg aattttatat tatcccgcgc agtcaggaaa gtaaatgtaa tgaatatatc 540
aacaacggcc tg 552
Claims (5)
1.一种CTB/LTB多肽与VP8的融合蛋白，其特征在于，所述融合蛋白为（1）CTB1-VP8 P[8]，其氨基酸序列如SEQ ID NO:4所示；或所述融合蛋白为（2）LTB1-VP8 P[8]，其氨基酸序列如SEQ ID NO:5所示。
2.一种轮状病毒疫苗，其特征在于，包含如权利要求1所述的融合蛋白。
3.根据权利要求2所述的轮状病毒疫苗，其特征在于，所述融合蛋白为10-100微克。
4.根据权利要求3所述的轮状病毒疫苗，其特征在于，还包含医学上可适用的佐剂。
5.根据权利要求4所述的轮状病毒疫苗，其特征在于，所述融合蛋白和佐剂体积比为1：1。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210355800.XA CN114805603B (zh) | 2022-04-06 | 2022-04-06 | 一种ctb/ltb多肽与vp8的融合蛋白及轮状病毒疫苗 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210355800.XA CN114805603B (zh) | 2022-04-06 | 2022-04-06 | 一种ctb/ltb多肽与vp8的融合蛋白及轮状病毒疫苗 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114805603A CN114805603A (zh) | 2022-07-29 |
| CN114805603B true CN114805603B (zh) | 2024-03-12 |
Family
ID=82533078
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210355800.XA Active CN114805603B (zh) | 2022-04-06 | 2022-04-06 | 一种ctb/ltb多肽与vp8的融合蛋白及轮状病毒疫苗 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114805603B (zh) |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103319604A (zh) * | 2013-06-18 | 2013-09-25 | 黑龙江八一农垦大学 | 一种人轮状病毒△vp8*亚单位重组蛋白及其应用 |
| CN105085639A (zh) * | 2014-05-21 | 2015-11-25 | 厦门大学 | 截短的轮状病毒vp8蛋白及其用途 |
| WO2018055535A2 (en) * | 2016-09-21 | 2018-03-29 | The Governors Of The University Of Alberta | Hepatitis c virus immunogenic compositions and methods of use thereof |
| CN113735945A (zh) * | 2018-04-13 | 2021-12-03 | 成都迈科康生物科技有限公司 | 一种人轮状病毒vp8重组蛋白以及利用该vp8重组蛋白的人用轮状病毒疫苗 |
-
2022
- 2022-04-06 CN CN202210355800.XA patent/CN114805603B/zh active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103319604A (zh) * | 2013-06-18 | 2013-09-25 | 黑龙江八一农垦大学 | 一种人轮状病毒△vp8*亚单位重组蛋白及其应用 |
| CN105085639A (zh) * | 2014-05-21 | 2015-11-25 | 厦门大学 | 截短的轮状病毒vp8蛋白及其用途 |
| WO2018055535A2 (en) * | 2016-09-21 | 2018-03-29 | The Governors Of The University Of Alberta | Hepatitis c virus immunogenic compositions and methods of use thereof |
| CN113735945A (zh) * | 2018-04-13 | 2021-12-03 | 成都迈科康生物科技有限公司 | 一种人轮状病毒vp8重组蛋白以及利用该vp8重组蛋白的人用轮状病毒疫苗 |
Non-Patent Citations (2)
| Title |
|---|
| "cholera enterotoxin B-subunit [Vibrio cholerae]，AAD51360.1";Shin, H.J.，等;NCBI GenBank;CDS * |
| Ochi,S.等."heat-labile enterotoxin LT subunit B [Escherichia coli]，WP_015675362.1".NCBI GenBank.2019,CDS. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114805603A (zh) | 2022-07-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102203252B (zh) | 基于鞭毛蛋白的新型免疫佐剂化合物及其用途 | |
| KR101100237B1 (ko) | 경구용 재조합 헬리코박터 파일로리 백신 및 그의 제조방법 | |
| CN108586618B (zh) | 一种猪流行性腹泻亚单位疫苗的制备及应用 | |
| WO2015176586A1 (zh) | 截短的轮状病毒vp8蛋白及其用途 | |
| EP1904626B1 (en) | Chimeric empty viral-like particles derived from the infectious bursal disease virus (ibdv), process for their production and applications | |
| NZ329833A (en) | Proteins useful for increasing the immunogenicity of an antigen and nucleotide sequences coding such proteins | |
| US5238823A (en) | Interleukin-2-leukotoxin gene fusions and uses thereof | |
| CN106928373B (zh) | 一种猪支原体肺炎多表位黏膜疫苗 | |
| CN109678968B (zh) | 猪肺炎支原体亚单位疫苗及其制备方法与应用 | |
| CN111548395A (zh) | 一种口蹄疫病毒二价多表位重组病毒样颗粒及其应用 | |
| CN111138551A (zh) | 一种预防金黄色葡萄球菌和白色念珠菌感染的atm融合蛋白的制备及应用 | |
| US5273889A (en) | Gamma-iterferon-leukotoxin gene fusions and uses thereof | |
| CN111607605B (zh) | 一种多价表位和亚单位疫苗的构建方法 | |
| CN114805603B (zh) | 一种ctb/ltb多肽与vp8的融合蛋白及轮状病毒疫苗 | |
| Shivachandra et al. | Immunogenicity of recombinant Omp16 protein of Pasteurella multocida B: 2 in mouse model | |
| CN110257405B (zh) | 牛支原体乙醇脱氢酶基因及其编码蛋白与应用 | |
| CN109369809B (zh) | 多表位抗原及其制备方法与在制备防治鹦鹉热衣原体感染的药物中的应用 | |
| CN116496406B (zh) | 激活tlr-5的幽门螺旋杆菌融合抗原及其应用 | |
| Xie et al. | Shuffling of pig interleukin-2 gene and its enhancing of immunity in mice to Pasteurella multocida vaccine | |
| US20030165526A1 (en) | Vaccines and agents for inducing immunity against rickettsial diseases, and associated preventative therapy | |
| CN114437236A (zh) | 一种重组非洲猪瘟病毒多表位融合蛋白、制备及其应用 | |
| KR101511800B1 (ko) | Ａ군 베타 용혈성 스트렙토코커스에 대한 백신 및 이의 수득 방법 | |
| JP6902804B2 (ja) | Ｂ型肝炎ウイルス様粒子をアジュバントとして含むワクチン組成物 | |
| CN111850003A (zh) | 一种重组表达的多杀性巴氏杆菌硫胺素周质结合蛋白及应用 | |
| WO2019175477A1 (en) | A subunit vaccine against porcine post-weaning diarrhoea |
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 |