CN107224575A - Composition of dairy cow staphylococcus aureus mastitis subunit vaccine and preparation method and application thereof - Google Patents
Composition of dairy cow staphylococcus aureus mastitis subunit vaccine and preparation method and application thereof Download PDFInfo
- Publication number
- CN107224575A CN107224575A CN201710128121.8A CN201710128121A CN107224575A CN 107224575 A CN107224575 A CN 107224575A CN 201710128121 A CN201710128121 A CN 201710128121A CN 107224575 A CN107224575 A CN 107224575A
- Authority
- CN
- China
- Prior art keywords
- albumen
- pvl
- composition
- hemolysin
- protein
- 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
- 239000000203 mixture Substances 0.000 title claims abstract description 85
- 241000191967 Staphylococcus aureus Species 0.000 title claims abstract description 26
- 208000004396 mastitis Diseases 0.000 title claims abstract description 22
- 229940031626 subunit vaccine Drugs 0.000 title claims abstract description 16
- 238000002360 preparation method Methods 0.000 title claims abstract description 12
- 235000013365 dairy product Nutrition 0.000 title claims abstract description 10
- 101710092462 Alpha-hemolysin Proteins 0.000 claims abstract description 43
- 101710124951 Phospholipase C Proteins 0.000 claims abstract description 34
- 241000283690 Bos taurus Species 0.000 claims abstract description 27
- 239000002671 adjuvant Substances 0.000 claims abstract description 22
- 239000007788 liquid Substances 0.000 claims abstract description 22
- RTKIYNMVFMVABJ-UHFFFAOYSA-L thimerosal Chemical compound [Na+].CC[Hg]SC1=CC=CC=C1C([O-])=O RTKIYNMVFMVABJ-UHFFFAOYSA-L 0.000 claims abstract description 13
- 229940033663 thimerosal Drugs 0.000 claims abstract description 13
- 239000000427 antigen Substances 0.000 claims abstract description 9
- 102000036639 antigens Human genes 0.000 claims abstract description 9
- 108091007433 antigens Proteins 0.000 claims abstract description 9
- 238000002156 mixing Methods 0.000 claims abstract description 9
- 239000008346 aqueous phase Substances 0.000 claims abstract description 7
- 229960005486 vaccine Drugs 0.000 claims description 26
- 235000013336 milk Nutrition 0.000 claims description 23
- 239000008267 milk Substances 0.000 claims description 23
- 210000004080 milk Anatomy 0.000 claims description 23
- 230000005847 immunogenicity Effects 0.000 claims description 9
- 230000002949 hemolytic effect Effects 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 206010018910 Haemolysis Diseases 0.000 claims description 6
- 230000008588 hemolysis Effects 0.000 claims description 6
- 206010064571 Gene mutation Diseases 0.000 claims description 5
- FFRBMBIXVSCUFS-UHFFFAOYSA-N 2,4-dinitro-1-naphthol Chemical compound C1=CC=C2C(O)=C([N+]([O-])=O)C=C([N+]([O-])=O)C2=C1 FFRBMBIXVSCUFS-UHFFFAOYSA-N 0.000 claims description 4
- 239000003755 preservative agent Substances 0.000 claims description 4
- 230000002335 preservative effect Effects 0.000 claims description 4
- 108010022355 Fibroins Proteins 0.000 claims description 3
- 230000001804 emulsifying effect Effects 0.000 claims description 3
- 108010068327 4-hydroxyphenylpyruvate dioxygenase Proteins 0.000 claims description 2
- 102100031673 Corneodesmosin Human genes 0.000 claims description 2
- 101710139375 Corneodesmosin Proteins 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 abstract description 23
- 239000003228 hemolysin Substances 0.000 abstract description 17
- 102000004169 proteins and genes Human genes 0.000 abstract description 13
- 238000006243 chemical reaction Methods 0.000 abstract description 9
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 7
- 239000012071 phase Substances 0.000 abstract description 3
- 239000000243 solution Substances 0.000 description 24
- 241000894006 Bacteria Species 0.000 description 19
- 239000000523 sample Substances 0.000 description 17
- 210000004027 cell Anatomy 0.000 description 12
- 230000004083 survival effect Effects 0.000 description 12
- 241000699666 Mus <mouse, genus> Species 0.000 description 11
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 10
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 9
- 230000029087 digestion Effects 0.000 description 9
- 231100000776 exotoxin Toxicity 0.000 description 9
- 239000002095 exotoxin Substances 0.000 description 9
- 238000001179 sorption measurement Methods 0.000 description 9
- 239000013612 plasmid Substances 0.000 description 8
- OTLLEIBWKHEHGU-UHFFFAOYSA-N 2-[5-[[5-(6-aminopurin-9-yl)-3,4-dihydroxyoxolan-2-yl]methoxy]-3,4-dihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy-3,5-dihydroxy-4-phosphonooxyhexanedioic acid Chemical compound C1=NC=2C(N)=NC=NC=2N1C(C(C1O)O)OC1COC1C(CO)OC(OC(C(O)C(OP(O)(O)=O)C(O)C(O)=O)C(O)=O)C(O)C1O OTLLEIBWKHEHGU-UHFFFAOYSA-N 0.000 description 7
- 108010006464 Hemolysin Proteins Proteins 0.000 description 7
- 238000005119 centrifugation Methods 0.000 description 7
- 231100000614 poison Toxicity 0.000 description 7
- 239000002574 poison Substances 0.000 description 7
- 235000018102 proteins Nutrition 0.000 description 7
- 230000003115 biocidal effect Effects 0.000 description 6
- 239000012149 elution buffer Substances 0.000 description 6
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 5
- 239000000872 buffer Substances 0.000 description 5
- 210000000170 cell membrane Anatomy 0.000 description 5
- 201000010099 disease Diseases 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 208000015181 infectious disease Diseases 0.000 description 5
- 238000011068 loading method Methods 0.000 description 5
- 238000005374 membrane filtration Methods 0.000 description 5
- 238000012797 qualification Methods 0.000 description 5
- 239000011780 sodium chloride Substances 0.000 description 5
- 108010005054 Deoxyribonuclease BamHI Proteins 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 238000013467 fragmentation Methods 0.000 description 4
- 238000006062 fragmentation reaction Methods 0.000 description 4
- 150000002460 imidazoles Chemical class 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 230000035772 mutation Effects 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 230000001681 protective effect Effects 0.000 description 4
- 230000006798 recombination Effects 0.000 description 4
- 238000005215 recombination Methods 0.000 description 4
- AJPJDKMHJJGVTQ-UHFFFAOYSA-M sodium dihydrogen phosphate Chemical compound [Na+].OP(O)([O-])=O AJPJDKMHJJGVTQ-UHFFFAOYSA-M 0.000 description 4
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 4
- 229910000162 sodium phosphate Inorganic materials 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 3
- 241000588724 Escherichia coli Species 0.000 description 3
- 238000000246 agarose gel electrophoresis Methods 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 239000007853 buffer solution Substances 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 230000004087 circulation Effects 0.000 description 3
- 238000004140 cleaning Methods 0.000 description 3
- 230000034994 death Effects 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000004945 emulsification Methods 0.000 description 3
- 239000003292 glue Substances 0.000 description 3
- 229940031551 inactivated vaccine Drugs 0.000 description 3
- 210000003622 mature neutrocyte Anatomy 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 238000001742 protein purification Methods 0.000 description 3
- 210000002966 serum Anatomy 0.000 description 3
- 239000003053 toxin Substances 0.000 description 3
- 231100000765 toxin Toxicity 0.000 description 3
- 108010038745 tryptophylglycine Proteins 0.000 description 3
- 230000003612 virological effect Effects 0.000 description 3
- 230000001018 virulence Effects 0.000 description 3
- 239000002699 waste material Substances 0.000 description 3
- RTFWCVDISAMGEQ-SRVKXCTJSA-N Asn-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](CC(=O)N)N RTFWCVDISAMGEQ-SRVKXCTJSA-N 0.000 description 2
- JILRMFFFCHUUTJ-ACZMJKKPSA-N Gln-Ser-Ser Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O JILRMFFFCHUUTJ-ACZMJKKPSA-N 0.000 description 2
- 241000880493 Leptailurus serval Species 0.000 description 2
- 241001465754 Metazoa Species 0.000 description 2
- 241000191940 Staphylococcus Species 0.000 description 2
- 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 2
- 102000014384 Type C Phospholipases Human genes 0.000 description 2
- 108010079194 Type C Phospholipases Proteins 0.000 description 2
- 235000001014 amino acid Nutrition 0.000 description 2
- 108010077245 asparaginyl-proline Proteins 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 235000010633 broth Nutrition 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000005859 coupling reaction Methods 0.000 description 2
- 125000004122 cyclic group Chemical group 0.000 description 2
- 238000011033 desalting Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 238000011067 equilibration Methods 0.000 description 2
- 210000003743 erythrocyte Anatomy 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 108010028295 histidylhistidine Proteins 0.000 description 2
- 230000006801 homologous recombination Effects 0.000 description 2
- 238000002744 homologous recombination Methods 0.000 description 2
- 230000002779 inactivation Effects 0.000 description 2
- 230000001939 inductive effect Effects 0.000 description 2
- 108010044374 isoleucyl-tyrosine Proteins 0.000 description 2
- 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 2
- 239000000463 material Substances 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 238000003672 processing method Methods 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 238000002560 therapeutic procedure Methods 0.000 description 2
- 239000000304 virulence factor Substances 0.000 description 2
- 230000007923 virulence factor Effects 0.000 description 2
- 239000011534 wash buffer Substances 0.000 description 2
- PKOHVHWNGUHYRE-ZFWWWQNUSA-N (2s)-1-[2-[[(2s)-2-amino-3-(1h-indol-3-yl)propanoyl]amino]acetyl]pyrrolidine-2-carboxylic acid Chemical compound O=C([C@H](CC=1C2=CC=CC=C2NC=1)N)NCC(=O)N1CCC[C@H]1C(O)=O PKOHVHWNGUHYRE-ZFWWWQNUSA-N 0.000 description 1
- INISVPAOPMKHCO-WZYQIZRJSA-N (2s)-6-amino-2-[[(2s)-2-[[(2s)-2-[[(2s)-2-[[(2s)-1-[(2s,3r)-2-amino-3-hydroxybutanoyl]pyrrolidine-2-carbonyl]amino]-3-methylbutanoyl]amino]-3-hydroxypropanoyl]amino]-4-carboxybutanoyl]amino]hexanoic acid Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@H](C(C)C)NC(=O)[C@@H]1CCCN1C(=O)[C@@H](N)[C@@H](C)O INISVPAOPMKHCO-WZYQIZRJSA-N 0.000 description 1
- IDOQDZANRZQBTP-UHFFFAOYSA-N 2-[2-(2,4,4-trimethylpentan-2-yl)phenoxy]ethanol Chemical compound CC(C)(C)CC(C)(C)C1=CC=CC=C1OCCO IDOQDZANRZQBTP-UHFFFAOYSA-N 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- JBGSZRYCXBPWGX-BQBZGAKWSA-N Ala-Arg-Gly Chemical compound OC(=O)CNC(=O)[C@@H](NC(=O)[C@@H](N)C)CCCN=C(N)N JBGSZRYCXBPWGX-BQBZGAKWSA-N 0.000 description 1
- WUHJHHGYVVJMQE-BJDJZHNGSA-N Ala-Leu-Ile Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WUHJHHGYVVJMQE-BJDJZHNGSA-N 0.000 description 1
- KQESEZXHYOUIIM-CQDKDKBSSA-N Ala-Lys-Tyr Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O KQESEZXHYOUIIM-CQDKDKBSSA-N 0.000 description 1
- NLOMBWNGESDVJU-GUBZILKMSA-N Ala-Met-Arg Chemical compound [H]N[C@@H](C)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O NLOMBWNGESDVJU-GUBZILKMSA-N 0.000 description 1
- AAWLEICNDUHIJM-MBLNEYKQSA-N Ala-Thr-His Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)O)NC(=O)[C@H](C)N)O AAWLEICNDUHIJM-MBLNEYKQSA-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
- 101710197219 Alpha-toxin Proteins 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
- OTUQSEPIIVBYEM-IHRRRGAJSA-N Arg-Asn-Tyr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O OTUQSEPIIVBYEM-IHRRRGAJSA-N 0.000 description 1
- FBLMOFHNVQBKRR-IHRRRGAJSA-N Arg-Asp-Tyr Chemical compound NC(N)=NCCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CC=C(O)C=C1 FBLMOFHNVQBKRR-IHRRRGAJSA-N 0.000 description 1
- SNBHMYQRNCJSOJ-CIUDSAMLSA-N Arg-Gln-Asn Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O SNBHMYQRNCJSOJ-CIUDSAMLSA-N 0.000 description 1
- ZPWMEWYQBWSGAO-ZJDVBMNYSA-N Arg-Thr-Thr Chemical compound [H]N[C@@H](CCCNC(N)=N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O ZPWMEWYQBWSGAO-ZJDVBMNYSA-N 0.000 description 1
- AYKKKGFJXIDYLX-ACZMJKKPSA-N Asn-Gln-Asn Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O AYKKKGFJXIDYLX-ACZMJKKPSA-N 0.000 description 1
- PQAIOUVVZCOLJK-FXQIFTODSA-N Asn-Gln-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CC(=O)N)N PQAIOUVVZCOLJK-FXQIFTODSA-N 0.000 description 1
- QNJIRRVTOXNGMH-GUBZILKMSA-N Asn-Gln-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](N)CC(N)=O QNJIRRVTOXNGMH-GUBZILKMSA-N 0.000 description 1
- BZMWJLLUAKSIMH-FXQIFTODSA-N Asn-Glu-Glu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCC(O)=O)C(O)=O BZMWJLLUAKSIMH-FXQIFTODSA-N 0.000 description 1
- JREOBWLIZLXRIS-GUBZILKMSA-N Asn-Glu-Leu Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(O)=O JREOBWLIZLXRIS-GUBZILKMSA-N 0.000 description 1
- ACKNRKFVYUVWAC-ZPFDUUQYSA-N Asn-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC(=O)N)N ACKNRKFVYUVWAC-ZPFDUUQYSA-N 0.000 description 1
- FTSAJSADJCMDHH-CIUDSAMLSA-N Asn-Lys-Asp Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)O)NC(=O)[C@H](CC(=O)N)N FTSAJSADJCMDHH-CIUDSAMLSA-N 0.000 description 1
- JWQWPRCDYWNVNM-ACZMJKKPSA-N Asn-Ser-Gln Chemical compound C(CC(=O)N)[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(=O)N)N JWQWPRCDYWNVNM-ACZMJKKPSA-N 0.000 description 1
- UGXYFDQFLVCDFC-CIUDSAMLSA-N Asn-Ser-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O UGXYFDQFLVCDFC-CIUDSAMLSA-N 0.000 description 1
- VLDRQOHCMKCXLY-SRVKXCTJSA-N Asn-Ser-Phe Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O VLDRQOHCMKCXLY-SRVKXCTJSA-N 0.000 description 1
- HPASIOLTWSNMFB-OLHMAJIHSA-N Asn-Thr-Asp Chemical compound [H]N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(O)=O)C(O)=O HPASIOLTWSNMFB-OLHMAJIHSA-N 0.000 description 1
- BEHQTVDBCLSCBY-CFMVVWHZSA-N Asn-Tyr-Ile 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]([C@@H](C)CC)C(O)=O BEHQTVDBCLSCBY-CFMVVWHZSA-N 0.000 description 1
- DATSKXOXPUAOLK-KKUMJFAQSA-N Asn-Tyr-Leu 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(C)C)C(O)=O DATSKXOXPUAOLK-KKUMJFAQSA-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
- CYCKJEFVFNRWEZ-UGYAYLCHSA-N Asp-Ile-Asn Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O CYCKJEFVFNRWEZ-UGYAYLCHSA-N 0.000 description 1
- RQHLMGCXCZUOGT-ZPFDUUQYSA-N Asp-Leu-Ile Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O RQHLMGCXCZUOGT-ZPFDUUQYSA-N 0.000 description 1
- DPNWSMBUYCLEDG-CIUDSAMLSA-N Asp-Lys-Ser Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O DPNWSMBUYCLEDG-CIUDSAMLSA-N 0.000 description 1
- ZXRQJQCXPSMNMR-XIRDDKMYSA-N Asp-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)O)N ZXRQJQCXPSMNMR-XIRDDKMYSA-N 0.000 description 1
- QSFHZPQUAAQHAQ-CIUDSAMLSA-N Asp-Ser-Leu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC(C)C)C(O)=O QSFHZPQUAAQHAQ-CIUDSAMLSA-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
- JSHWXQIZOCVWIA-ZKWXMUAHSA-N Asp-Ser-Val Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(O)=O JSHWXQIZOCVWIA-ZKWXMUAHSA-N 0.000 description 1
- MRYDJCIIVRXVGG-QEJZJMRPSA-N Asp-Trp-Glu Chemical compound [H]N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCC(O)=O)C(O)=O MRYDJCIIVRXVGG-QEJZJMRPSA-N 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 108090000397 Caspase 3 Proteins 0.000 description 1
- 102100029855 Caspase-3 Human genes 0.000 description 1
- 102100026550 Caspase-9 Human genes 0.000 description 1
- 108090000566 Caspase-9 Proteins 0.000 description 1
- 241001478240 Coccus Species 0.000 description 1
- 208000035473 Communicable disease Diseases 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 235000009161 Espostoa lanata Nutrition 0.000 description 1
- 240000001624 Espostoa lanata Species 0.000 description 1
- 108090000371 Esterases Proteins 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- WOACHWLUOFZLGJ-GUBZILKMSA-N Gln-Arg-Gln Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(N)=O)C(O)=O WOACHWLUOFZLGJ-GUBZILKMSA-N 0.000 description 1
- WMOMPXKOKASNBK-PEFMBERDSA-N Gln-Asn-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WMOMPXKOKASNBK-PEFMBERDSA-N 0.000 description 1
- CKNUKHBRCSMKMO-XHNCKOQMSA-N Gln-Asn-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)N)NC(=O)[C@H](CCC(=O)N)N)C(=O)O CKNUKHBRCSMKMO-XHNCKOQMSA-N 0.000 description 1
- KQOPMGBHNQBCEL-HVTMNAMFSA-N Gln-His-Ile Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KQOPMGBHNQBCEL-HVTMNAMFSA-N 0.000 description 1
- FTIJVMLAGRAYMJ-MNXVOIDGSA-N Gln-Ile-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCC(N)=O FTIJVMLAGRAYMJ-MNXVOIDGSA-N 0.000 description 1
- TWIAMTNJOMRDAK-GUBZILKMSA-N Gln-Lys-Asp Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(O)=O TWIAMTNJOMRDAK-GUBZILKMSA-N 0.000 description 1
- NHMRJKKAVMENKJ-WDCWCFNPSA-N Gln-Thr-Leu Chemical compound [H]N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(C)C)C(O)=O NHMRJKKAVMENKJ-WDCWCFNPSA-N 0.000 description 1
- WPJDPEOQUIXXOY-AVGNSLFASA-N Gln-Tyr-Asn Chemical compound C1=CC(=CC=C1C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)O)NC(=O)[C@H](CCC(=O)N)N)O WPJDPEOQUIXXOY-AVGNSLFASA-N 0.000 description 1
- IRXNJYPKBVERCW-DCAQKATOSA-N Glu-Leu-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O IRXNJYPKBVERCW-DCAQKATOSA-N 0.000 description 1
- JDUKCSSHWNIQQZ-IHRRRGAJSA-N Glu-Phe-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CCC(O)=O)C(O)=O JDUKCSSHWNIQQZ-IHRRRGAJSA-N 0.000 description 1
- JZJGEKDPWVJOLD-QEWYBTABSA-N Glu-Phe-Ile Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JZJGEKDPWVJOLD-QEWYBTABSA-N 0.000 description 1
- 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 1
- WXONSNSSBYQGNN-AVGNSLFASA-N Glu-Ser-Tyr Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O WXONSNSSBYQGNN-AVGNSLFASA-N 0.000 description 1
- RGJKYNUINKGPJN-RWRJDSDZSA-N Glu-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](CCC(=O)O)N RGJKYNUINKGPJN-RWRJDSDZSA-N 0.000 description 1
- LZEUDRYSAZAJIO-AUTRQRHGSA-N Glu-Val-Glu Chemical compound [H]N[C@@H](CCC(O)=O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O LZEUDRYSAZAJIO-AUTRQRHGSA-N 0.000 description 1
- MFVQGXGQRIXBPK-WDSKDSINSA-N Gly-Ala-Glu Chemical compound NCC(=O)N[C@@H](C)C(=O)N[C@@H](CCC(O)=O)C(O)=O MFVQGXGQRIXBPK-WDSKDSINSA-N 0.000 description 1
- XUDLUKYPXQDCRX-BQBZGAKWSA-N Gly-Arg-Asn Chemical compound [H]NCC(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(N)=O)C(O)=O XUDLUKYPXQDCRX-BQBZGAKWSA-N 0.000 description 1
- GGEJHJIXRBTJPD-BYPYZUCNSA-N Gly-Asn-Gly Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)NCC(O)=O GGEJHJIXRBTJPD-BYPYZUCNSA-N 0.000 description 1
- BGVYNAQWHSTTSP-BYULHYEWSA-N Gly-Asn-Ile Chemical compound [H]NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O BGVYNAQWHSTTSP-BYULHYEWSA-N 0.000 description 1
- XCLCVBYNGXEVDU-WHFBIAKZSA-N Gly-Asn-Ser Chemical compound NCC(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CO)C(O)=O XCLCVBYNGXEVDU-WHFBIAKZSA-N 0.000 description 1
- PMNHJLASAAWELO-FOHZUACHSA-N Gly-Asp-Thr Chemical compound [H]NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O PMNHJLASAAWELO-FOHZUACHSA-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
- AYBKPDHHVADEDA-YUMQZZPRSA-N Gly-His-Asn Chemical compound [H]NCC(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC(N)=O)C(O)=O AYBKPDHHVADEDA-YUMQZZPRSA-N 0.000 description 1
- ITZOBNKQDZEOCE-NHCYSSNCSA-N Gly-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)CN ITZOBNKQDZEOCE-NHCYSSNCSA-N 0.000 description 1
- IUZGUFAJDBHQQV-YUMQZZPRSA-N Gly-Leu-Asn Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O IUZGUFAJDBHQQV-YUMQZZPRSA-N 0.000 description 1
- LLZXNUUIBOALNY-QWRGUYRKSA-N Gly-Leu-Lys Chemical compound NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@H](C(O)=O)CCCCN LLZXNUUIBOALNY-QWRGUYRKSA-N 0.000 description 1
- MTBIKIMYHUWBRX-QWRGUYRKSA-N Gly-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)CN MTBIKIMYHUWBRX-QWRGUYRKSA-N 0.000 description 1
- IBYOLNARKHMLBG-WHOFXGATSA-N Gly-Phe-Ile Chemical compound CC[C@H](C)[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=CC=C1 IBYOLNARKHMLBG-WHOFXGATSA-N 0.000 description 1
- YOBGUCWZPXJHTN-BQBZGAKWSA-N Gly-Ser-Arg Chemical compound NCC(=O)N[C@@H](CO)C(=O)N[C@H](C(O)=O)CCCN=C(N)N YOBGUCWZPXJHTN-BQBZGAKWSA-N 0.000 description 1
- POJJAZJHBGXEGM-YUMQZZPRSA-N Gly-Ser-Lys Chemical compound C(CCN)C[C@@H](C(=O)O)NC(=O)[C@H](CO)NC(=O)CN POJJAZJHBGXEGM-YUMQZZPRSA-N 0.000 description 1
- PYFIQROSWQERAS-LBPRGKRZSA-N Gly-Trp-Gly Chemical compound C1=CC=C2C(C[C@H](NC(=O)CN)C(=O)NCC(O)=O)=CNC2=C1 PYFIQROSWQERAS-LBPRGKRZSA-N 0.000 description 1
- PNUFMLXHOLFRLD-KBPBESRZSA-N Gly-Tyr-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 PNUFMLXHOLFRLD-KBPBESRZSA-N 0.000 description 1
- DNAZKGFYFRGZIH-QWRGUYRKSA-N Gly-Tyr-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CC=C(O)C=C1 DNAZKGFYFRGZIH-QWRGUYRKSA-N 0.000 description 1
- AFMOTCMSEBITOE-YEPSODPASA-N Gly-Val-Thr Chemical compound NCC(=O)N[C@@H](C(C)C)C(=O)N[C@@H]([C@@H](C)O)C(O)=O AFMOTCMSEBITOE-YEPSODPASA-N 0.000 description 1
- MAABHGXCIBEYQR-XVYDVKMFSA-N His-Asn-Ala Chemical compound C[C@@H](C(=O)O)NC(=O)[C@H](CC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N MAABHGXCIBEYQR-XVYDVKMFSA-N 0.000 description 1
- LMMPTUVWHCFTOT-GARJFASQSA-N His-Asp-Pro Chemical compound C1C[C@@H](N(C1)C(=O)[C@H](CC(=O)O)NC(=O)[C@H](CC2=CN=CN2)N)C(=O)O LMMPTUVWHCFTOT-GARJFASQSA-N 0.000 description 1
- LPZUKJALYGXBIE-SRVKXCTJSA-N His-Gln-Met Chemical compound CSCC[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CN=CN1)N LPZUKJALYGXBIE-SRVKXCTJSA-N 0.000 description 1
- HQKADFMLECZIQJ-HVTMNAMFSA-N His-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC1=CN=CN1)N HQKADFMLECZIQJ-HVTMNAMFSA-N 0.000 description 1
- STOOMQFEJUVAKR-KKUMJFAQSA-N His-His-His Chemical compound C([C@H](N)C(=O)N[C@@H](CC=1N=CNC=1)C(=O)N[C@@H](CC=1N=CNC=1)C(O)=O)C1=CNC=N1 STOOMQFEJUVAKR-KKUMJFAQSA-N 0.000 description 1
- 108010093488 His-His-His-His-His-His Proteins 0.000 description 1
- QMUHTRISZMFKAY-MXAVVETBSA-N His-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CN=CN1)N QMUHTRISZMFKAY-MXAVVETBSA-N 0.000 description 1
- LDFWDDVELNOGII-MXAVVETBSA-N His-Lys-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](CC1=CN=CN1)N LDFWDDVELNOGII-MXAVVETBSA-N 0.000 description 1
- UMYZBHKAVTXWIW-GMOBBJLQSA-N Ile-Asp-Arg Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CC(=O)O)C(=O)N[C@@H](CCCN=C(N)N)C(=O)O)N UMYZBHKAVTXWIW-GMOBBJLQSA-N 0.000 description 1
- DCQMJRSOGCYKTR-GHCJXIJMSA-N Ile-Asp-Ser Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CO)C(O)=O DCQMJRSOGCYKTR-GHCJXIJMSA-N 0.000 description 1
- BEWFWZRGBDVXRP-PEFMBERDSA-N Ile-Glu-Asn Chemical compound [H]N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O BEWFWZRGBDVXRP-PEFMBERDSA-N 0.000 description 1
- KFVUBLZRFSVDGO-BYULHYEWSA-N Ile-Gly-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(O)=O KFVUBLZRFSVDGO-BYULHYEWSA-N 0.000 description 1
- CDGLBYSAZFIIJO-RCOVLWMOSA-N Ile-Gly-Gly Chemical compound CC[C@H](C)[C@H]([NH3+])C(=O)NCC(=O)NCC([O-])=O CDGLBYSAZFIIJO-RCOVLWMOSA-N 0.000 description 1
- UAQSZXGJGLHMNV-XEGUGMAKSA-N Ile-Gly-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)NCC(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N UAQSZXGJGLHMNV-XEGUGMAKSA-N 0.000 description 1
- SVBAHOMTJRFSIC-SXTJYALSSA-N Ile-Ile-Asn Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(=O)N)C(=O)O)N SVBAHOMTJRFSIC-SXTJYALSSA-N 0.000 description 1
- WLRJHVNFGAOYPS-HJPIBITLSA-N Ile-Ser-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N WLRJHVNFGAOYPS-HJPIBITLSA-N 0.000 description 1
- WCNWGAUZWWSYDG-SVSWQMSJSA-N Ile-Thr-Ser Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)O)N WCNWGAUZWWSYDG-SVSWQMSJSA-N 0.000 description 1
- DGTOKVBDZXJHNZ-WZLNRYEVSA-N Ile-Thr-Tyr Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O)N DGTOKVBDZXJHNZ-WZLNRYEVSA-N 0.000 description 1
- BCISUQVFDGYZBO-QSFUFRPTSA-N Ile-Val-Asp Chemical compound CC[C@H](C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CC(O)=O BCISUQVFDGYZBO-QSFUFRPTSA-N 0.000 description 1
- 206010061218 Inflammation Diseases 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
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 1
- IWTBYNQNAPECCS-AVGNSLFASA-N Leu-Glu-His Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@H](C(O)=O)CC1=CN=CN1 IWTBYNQNAPECCS-AVGNSLFASA-N 0.000 description 1
- APFJUBGRZGMQFF-QWRGUYRKSA-N Leu-Gly-Lys Chemical compound CC(C)C[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CCCCN APFJUBGRZGMQFF-QWRGUYRKSA-N 0.000 description 1
- UCDHVOALNXENLC-KBPBESRZSA-N Leu-Gly-Tyr Chemical compound CC(C)C[C@H]([NH3+])C(=O)NCC(=O)N[C@H](C([O-])=O)CC1=CC=C(O)C=C1 UCDHVOALNXENLC-KBPBESRZSA-N 0.000 description 1
- LKXANTUNFMVCNF-IHPCNDPISA-N Leu-His-Trp Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CNC2=C1C=CC=C2)C(O)=O LKXANTUNFMVCNF-IHPCNDPISA-N 0.000 description 1
- KUIDCYNIEJBZBU-AJNGGQMLSA-N Leu-Ile-Leu Chemical compound [H]N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O KUIDCYNIEJBZBU-AJNGGQMLSA-N 0.000 description 1
- DRWMRVFCKKXHCH-BZSNNMDCSA-N Leu-Phe-Leu Chemical compound CC(C)C[C@H]([NH3+])C(=O)N[C@H](C(=O)N[C@@H](CC(C)C)C([O-])=O)CC1=CC=CC=C1 DRWMRVFCKKXHCH-BZSNNMDCSA-N 0.000 description 1
- FYPWFNKQVVEELI-ULQDDVLXSA-N Leu-Phe-Val Chemical compound CC(C)C[C@H](N)C(=O)N[C@H](C(=O)N[C@@H](C(C)C)C(O)=O)CC1=CC=CC=C1 FYPWFNKQVVEELI-ULQDDVLXSA-N 0.000 description 1
- PWPBLZXWFXJFHE-RHYQMDGZSA-N Leu-Pro-Thr Chemical compound CC(C)C[C@H](N)C(=O)N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)O)C(O)=O PWPBLZXWFXJFHE-RHYQMDGZSA-N 0.000 description 1
- VDIARPPNADFEAV-WEDXCCLWSA-N Leu-Thr-Gly Chemical compound CC(C)C[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)NCC(O)=O VDIARPPNADFEAV-WEDXCCLWSA-N 0.000 description 1
- BTEMNFBEAAOGBR-BZSNNMDCSA-N Leu-Tyr-Lys Chemical compound CC(C)C[C@@H](C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)N[C@@H](CCCCN)C(=O)O)N BTEMNFBEAAOGBR-BZSNNMDCSA-N 0.000 description 1
- 108010014603 Leukocidins Proteins 0.000 description 1
- FZIJIFCXUCZHOL-CIUDSAMLSA-N Lys-Ala-Ala Chemical compound OC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](N)CCCCN FZIJIFCXUCZHOL-CIUDSAMLSA-N 0.000 description 1
- YKIRNDPUWONXQN-GUBZILKMSA-N Lys-Asn-Gln Chemical compound C(CCN)C[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CCC(=O)N)C(=O)O)N YKIRNDPUWONXQN-GUBZILKMSA-N 0.000 description 1
- ZQCVMVCVPFYXHZ-SRVKXCTJSA-N Lys-Asn-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@H](C(O)=O)CCCCN ZQCVMVCVPFYXHZ-SRVKXCTJSA-N 0.000 description 1
- KWUKZRFFKPLUPE-HJGDQZAQSA-N Lys-Asp-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KWUKZRFFKPLUPE-HJGDQZAQSA-N 0.000 description 1
- FHIAJWBDZVHLAH-YUMQZZPRSA-N Lys-Gly-Ser Chemical compound NCCCC[C@H](N)C(=O)NCC(=O)N[C@@H](CO)C(O)=O FHIAJWBDZVHLAH-YUMQZZPRSA-N 0.000 description 1
- HQXSFFSLXFHWOX-IXOXFDKPSA-N Lys-His-Thr Chemical compound C[C@H]([C@@H](C(=O)O)NC(=O)[C@H](CC1=CN=CN1)NC(=O)[C@H](CCCCN)N)O HQXSFFSLXFHWOX-IXOXFDKPSA-N 0.000 description 1
- KYNNSEJZFVCDIV-ZPFDUUQYSA-N Lys-Ile-Asn Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(N)=O)C(O)=O KYNNSEJZFVCDIV-ZPFDUUQYSA-N 0.000 description 1
- QOJDBRUCOXQSSK-AJNGGQMLSA-N Lys-Ile-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CCCCN)C(O)=O QOJDBRUCOXQSSK-AJNGGQMLSA-N 0.000 description 1
- PRSBSVAVOQOAMI-BJDJZHNGSA-N Lys-Ile-Ser Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H]([C@@H](C)CC)NC(=O)[C@@H](N)CCCCN PRSBSVAVOQOAMI-BJDJZHNGSA-N 0.000 description 1
- NCZIQZYZPUPMKY-PPCPHDFISA-N Lys-Ile-Thr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(O)=O NCZIQZYZPUPMKY-PPCPHDFISA-N 0.000 description 1
- WVJNGSFKBKOKRV-AJNGGQMLSA-N Lys-Leu-Ile Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O WVJNGSFKBKOKRV-AJNGGQMLSA-N 0.000 description 1
- BXPHMHQHYHILBB-BZSNNMDCSA-N Lys-Lys-Tyr Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O BXPHMHQHYHILBB-BZSNNMDCSA-N 0.000 description 1
- WKUXWMWQTOYTFI-SRVKXCTJSA-N Lys-Met-Gln Chemical compound CSCC[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)O)NC(=O)[C@H](CCCCN)N WKUXWMWQTOYTFI-SRVKXCTJSA-N 0.000 description 1
- QLFAPXUXEBAWEK-NHCYSSNCSA-N Lys-Val-Asp Chemical compound [H]N[C@@H](CCCCN)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(O)=O)C(O)=O QLFAPXUXEBAWEK-NHCYSSNCSA-N 0.000 description 1
- NYTDJEZBAAFLLG-IHRRRGAJSA-N Lys-Val-Lys Chemical compound NCCCC[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(O)=O NYTDJEZBAAFLLG-IHRRRGAJSA-N 0.000 description 1
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- FRWZTWWOORIIBA-FXQIFTODSA-N Met-Asn-Asn Chemical compound CSCC[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)N)C(=O)O)N FRWZTWWOORIIBA-FXQIFTODSA-N 0.000 description 1
- SQUTUWHAAWJYES-GUBZILKMSA-N Met-Asp-Arg Chemical compound [H]N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O SQUTUWHAAWJYES-GUBZILKMSA-N 0.000 description 1
- FVKRQMQQFGBXHV-QXEWZRGKSA-N Met-Asp-Val Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](C(C)C)C(O)=O FVKRQMQQFGBXHV-QXEWZRGKSA-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
- WWWGMQHQSAUXBU-BQBZGAKWSA-N Met-Gly-Asn Chemical compound CSCC[C@H](N)C(=O)NCC(=O)N[C@H](C(O)=O)CC(N)=O WWWGMQHQSAUXBU-BQBZGAKWSA-N 0.000 description 1
- LXCSZPUQKMTXNW-BQBZGAKWSA-N Met-Ser-Gly Chemical compound CSCC[C@H](N)C(=O)N[C@@H](CO)C(=O)NCC(O)=O LXCSZPUQKMTXNW-BQBZGAKWSA-N 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 206010056720 Muscle mass Diseases 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
- 108010078471 Panton-Valentine leukocidin Proteins 0.000 description 1
- 101710161551 Pectate lyase 3 Proteins 0.000 description 1
- LXVFHIBXOWJTKZ-BZSNNMDCSA-N Phe-Asn-Tyr Chemical compound N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(C=C1)O)C(=O)O LXVFHIBXOWJTKZ-BZSNNMDCSA-N 0.000 description 1
- DJPXNKUDJKGQEE-BZSNNMDCSA-N Phe-Asp-Phe Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O DJPXNKUDJKGQEE-BZSNNMDCSA-N 0.000 description 1
- MFQXSDWKUXTOPZ-DZKIICNBSA-N Phe-Gln-Val Chemical compound CC(C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)N)NC(=O)[C@H](CC1=CC=CC=C1)N MFQXSDWKUXTOPZ-DZKIICNBSA-N 0.000 description 1
- WEMYTDDMDBLPMI-DKIMLUQUSA-N Phe-Ile-Lys Chemical compound CC[C@H](C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)O)NC(=O)[C@H](CC1=CC=CC=C1)N WEMYTDDMDBLPMI-DKIMLUQUSA-N 0.000 description 1
- IPVPGAADZXRZSH-RNXOBYDBSA-N Phe-Tyr-Trp 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](CC1=CNC2=C1C=CC=C2)C(O)=O IPVPGAADZXRZSH-RNXOBYDBSA-N 0.000 description 1
- GOUWCZRDTWTODO-YDHLFZDLSA-N Phe-Val-Asn Chemical compound [H]N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(N)=O)C(O)=O GOUWCZRDTWTODO-YDHLFZDLSA-N 0.000 description 1
- AHXPYZRZRMQOAU-QXEWZRGKSA-N Pro-Asn-Val Chemical compound CC(C)[C@H](NC(=O)[C@H](CC(N)=O)NC(=O)[C@@H]1CCCN1)C(O)=O AHXPYZRZRMQOAU-QXEWZRGKSA-N 0.000 description 1
- VZKBJNBZMZHKRC-XUXIUFHCSA-N Pro-Ile-Leu Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC(C)C)C(O)=O VZKBJNBZMZHKRC-XUXIUFHCSA-N 0.000 description 1
- CGSOWZUPLOKYOR-AVGNSLFASA-N Pro-Pro-Leu Chemical compound CC(C)C[C@@H](C(O)=O)NC(=O)[C@@H]1CCCN1C(=O)[C@H]1NCCC1 CGSOWZUPLOKYOR-AVGNSLFASA-N 0.000 description 1
- VEUACYMXJKXALX-IHRRRGAJSA-N Pro-Tyr-Ser Chemical compound [H]N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(O)=O VEUACYMXJKXALX-IHRRRGAJSA-N 0.000 description 1
- 101710179609 Probable pectin lyase C Proteins 0.000 description 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 description 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 description 1
- XVAUJOAYHWWNQF-ZLUOBGJFSA-N Ser-Asn-Ala Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](C)C(O)=O XVAUJOAYHWWNQF-ZLUOBGJFSA-N 0.000 description 1
- BYIROAKULFFTEK-CIUDSAMLSA-N Ser-Asp-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@H](CC(O)=O)NC(=O)[C@@H](N)CO BYIROAKULFFTEK-CIUDSAMLSA-N 0.000 description 1
- KJMOINFQVCCSDX-XKBZYTNZSA-N Ser-Gln-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(O)=O KJMOINFQVCCSDX-XKBZYTNZSA-N 0.000 description 1
- KDGARKCAKHBEDB-NKWVEPMBSA-N Ser-Gly-Pro Chemical compound C1C[C@@H](N(C1)C(=O)CNC(=O)[C@H](CO)N)C(=O)O KDGARKCAKHBEDB-NKWVEPMBSA-N 0.000 description 1
- HMRAQFJFTOLDKW-GUBZILKMSA-N Ser-His-Glu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CCC(O)=O)C(O)=O HMRAQFJFTOLDKW-GUBZILKMSA-N 0.000 description 1
- CLKKNZQUQMZDGD-SRVKXCTJSA-N Ser-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)[C@@H](N)CO)CC1=CN=CN1 CLKKNZQUQMZDGD-SRVKXCTJSA-N 0.000 description 1
- FPCGZYMRFFIYIH-CIUDSAMLSA-N Ser-Lys-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CO)C(O)=O FPCGZYMRFFIYIH-CIUDSAMLSA-N 0.000 description 1
- LRZLZIUXQBIWTB-KATARQTJSA-N Ser-Lys-Thr Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O LRZLZIUXQBIWTB-KATARQTJSA-N 0.000 description 1
- KZPRPBLHYMZIMH-MXAVVETBSA-N Ser-Phe-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O KZPRPBLHYMZIMH-MXAVVETBSA-N 0.000 description 1
- MQUZANJDFOQOBX-SRVKXCTJSA-N Ser-Phe-Ser Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CO)C(O)=O MQUZANJDFOQOBX-SRVKXCTJSA-N 0.000 description 1
- JCLAFVNDBJMLBC-JBDRJPRFSA-N Ser-Ser-Ile Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O JCLAFVNDBJMLBC-JBDRJPRFSA-N 0.000 description 1
- PURRNJBBXDDWLX-ZDLURKLDSA-N Ser-Thr-Gly Chemical compound C[C@H]([C@@H](C(=O)NCC(=O)O)NC(=O)[C@H](CO)N)O PURRNJBBXDDWLX-ZDLURKLDSA-N 0.000 description 1
- YEDSOSIKVUMIJE-DCAQKATOSA-N Ser-Val-Leu Chemical compound [H]N[C@@H](CO)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O YEDSOSIKVUMIJE-DCAQKATOSA-N 0.000 description 1
- 102100032889 Sortilin Human genes 0.000 description 1
- 206010041925 Staphylococcal infections Diseases 0.000 description 1
- 241000193985 Streptococcus agalactiae Species 0.000 description 1
- YLXAMFZYJTZXFH-OLHMAJIHSA-N Thr-Asn-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O YLXAMFZYJTZXFH-OLHMAJIHSA-N 0.000 description 1
- DIPIPFHFLPTCLK-LOKLDPHHSA-N Thr-Gln-Pro Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N1CCC[C@@H]1C(=O)O)N)O DIPIPFHFLPTCLK-LOKLDPHHSA-N 0.000 description 1
- LGNBRHZANHMZHK-NUMRIWBASA-N Thr-Glu-Asp Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CCC(=O)O)C(=O)N[C@@H](CC(=O)O)C(=O)O)N)O LGNBRHZANHMZHK-NUMRIWBASA-N 0.000 description 1
- SIMKLINEDYOTKL-MBLNEYKQSA-N Thr-His-Ala Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](C)C(=O)O)N)O SIMKLINEDYOTKL-MBLNEYKQSA-N 0.000 description 1
- YDWLCDQXLCILCZ-BWAGICSOSA-N Thr-His-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CNC=N1)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O YDWLCDQXLCILCZ-BWAGICSOSA-N 0.000 description 1
- KZURUCDWKDEAFZ-XVSYOHENSA-N Thr-Phe-Asn Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](CC1=CC=CC=C1)C(=O)N[C@@H](CC(=O)N)C(=O)O)N)O KZURUCDWKDEAFZ-XVSYOHENSA-N 0.000 description 1
- NQQMWWVVGIXUOX-SVSWQMSJSA-N Thr-Ser-Ile Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H]([C@@H](C)CC)C(O)=O NQQMWWVVGIXUOX-SVSWQMSJSA-N 0.000 description 1
- WPSKTVVMQCXPRO-BWBBJGPYSA-N Thr-Ser-Ser Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(O)=O WPSKTVVMQCXPRO-BWBBJGPYSA-N 0.000 description 1
- NDZYTIMDOZMECO-SHGPDSBTSA-N Thr-Thr-Ala Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C)C(O)=O NDZYTIMDOZMECO-SHGPDSBTSA-N 0.000 description 1
- GRIUMVXCJDKVPI-IZPVPAKOSA-N Thr-Thr-Tyr Chemical compound [H]N[C@@H]([C@@H](C)O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O GRIUMVXCJDKVPI-IZPVPAKOSA-N 0.000 description 1
- PWONLXBUSVIZPH-RHYQMDGZSA-N Thr-Val-Lys Chemical compound C[C@H]([C@@H](C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)O)N)O PWONLXBUSVIZPH-RHYQMDGZSA-N 0.000 description 1
- 229920004929 Triton X-114 Polymers 0.000 description 1
- YEGMNOHLZNGOCG-UBHSHLNASA-N Trp-Asn-Asn Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC(N)=O)C(O)=O YEGMNOHLZNGOCG-UBHSHLNASA-N 0.000 description 1
- WKQNLTQSCYXKQK-VFAJRCTISA-N Trp-Lys-Thr Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H]([C@@H](C)O)C(O)=O WKQNLTQSCYXKQK-VFAJRCTISA-N 0.000 description 1
- GFUOTIPYXKAPAH-BVSLBCMMSA-N Trp-Pro-Phe Chemical compound [H]N[C@@H](CC1=CNC2=C1C=CC=C2)C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC1=CC=CC=C1)C(O)=O GFUOTIPYXKAPAH-BVSLBCMMSA-N 0.000 description 1
- OOEUVMFKKZYSRX-LEWSCRJBSA-N Tyr-Ala-Pro Chemical compound C[C@@H](C(=O)N1CCC[C@@H]1C(=O)O)NC(=O)[C@H](CC2=CC=C(C=C2)O)N OOEUVMFKKZYSRX-LEWSCRJBSA-N 0.000 description 1
- NSTPFWRAIDTNGH-BZSNNMDCSA-N Tyr-Asn-Tyr Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CC1=CC=C(O)C=C1)C(O)=O NSTPFWRAIDTNGH-BZSNNMDCSA-N 0.000 description 1
- LOOCQRRBKZTPKO-AVGNSLFASA-N Tyr-Glu-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](N)CC1=CC=C(O)C=C1 LOOCQRRBKZTPKO-AVGNSLFASA-N 0.000 description 1
- IMXAAEFAIBRCQF-SIUGBPQLSA-N Tyr-Glu-Ile Chemical compound CC[C@H](C)[C@@H](C(=O)O)NC(=O)[C@H](CCC(=O)O)NC(=O)[C@H](CC1=CC=C(C=C1)O)N IMXAAEFAIBRCQF-SIUGBPQLSA-N 0.000 description 1
- UNUZEBFXGWVAOP-DZKIICNBSA-N Tyr-Glu-Val 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](C(C)C)C(O)=O UNUZEBFXGWVAOP-DZKIICNBSA-N 0.000 description 1
- PMDWYLVWHRTJIW-STQMWFEESA-N Tyr-Gly-Arg Chemical compound NC(N)=NCCC[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 PMDWYLVWHRTJIW-STQMWFEESA-N 0.000 description 1
- CDHQEOXPWBDFPL-QWRGUYRKSA-N Tyr-Gly-Asn Chemical compound NC(=O)C[C@@H](C(O)=O)NC(=O)CNC(=O)[C@@H](N)CC1=CC=C(O)C=C1 CDHQEOXPWBDFPL-QWRGUYRKSA-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
- KSCVLGXNQXKUAR-JYJNAYRXSA-N Tyr-Leu-Glu Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCC(O)=O)C(O)=O KSCVLGXNQXKUAR-JYJNAYRXSA-N 0.000 description 1
- BYAKMYBZADCNMN-JYJNAYRXSA-N Tyr-Lys-Gln Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(N)=O)C(O)=O BYAKMYBZADCNMN-JYJNAYRXSA-N 0.000 description 1
- ZPFLBLFITJCBTP-QWRGUYRKSA-N Tyr-Ser-Gly Chemical compound [H]N[C@@H](CC1=CC=C(O)C=C1)C(=O)N[C@@H](CO)C(=O)NCC(O)=O ZPFLBLFITJCBTP-QWRGUYRKSA-N 0.000 description 1
- AZSHAZJLOZQYAY-FXQIFTODSA-N Val-Ala-Ser Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C)C(=O)N[C@@H](CO)C(O)=O AZSHAZJLOZQYAY-FXQIFTODSA-N 0.000 description 1
- DBOXBUDEAJVKRE-LSJOCFKGSA-N Val-Asn-Val Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC(=O)N)C(=O)N[C@@H](C(C)C)C(=O)O)N DBOXBUDEAJVKRE-LSJOCFKGSA-N 0.000 description 1
- OXVPMZVGCAPFIG-BQFCYCMXSA-N Val-Gln-Trp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCC(=O)N)C(=O)N[C@@H](CC1=CNC2=CC=CC=C21)C(=O)O)N OXVPMZVGCAPFIG-BQFCYCMXSA-N 0.000 description 1
- XGJLNBNZNMVJRS-NRPADANISA-N Val-Glu-Ala Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(O)=O XGJLNBNZNMVJRS-NRPADANISA-N 0.000 description 1
- HQYVQDRYODWONX-DCAQKATOSA-N Val-His-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CC1=CN=CN1)C(=O)N[C@@H](CO)C(=O)O)N HQYVQDRYODWONX-DCAQKATOSA-N 0.000 description 1
- KTEZUXISLQTDDQ-NHCYSSNCSA-N Val-Lys-Asp Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CC(=O)O)C(=O)O)N KTEZUXISLQTDDQ-NHCYSSNCSA-N 0.000 description 1
- YMTOEGGOCHVGEH-IHRRRGAJSA-N Val-Lys-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCCCN)C(O)=O YMTOEGGOCHVGEH-IHRRRGAJSA-N 0.000 description 1
- ZXYPHBKIZLAQTL-QXEWZRGKSA-N Val-Pro-Asp Chemical compound CC(C)[C@@H](C(=O)N1CCC[C@H]1C(=O)N[C@@H](CC(=O)O)C(=O)O)N ZXYPHBKIZLAQTL-QXEWZRGKSA-N 0.000 description 1
- PZTZYZUTCPZWJH-FXQIFTODSA-N Val-Ser-Ser Chemical compound CC(C)[C@@H](C(=O)N[C@@H](CO)C(=O)N[C@@H](CO)C(=O)O)N PZTZYZUTCPZWJH-FXQIFTODSA-N 0.000 description 1
- YLBNZCJFSVJDRJ-KJEVXHAQSA-N Val-Thr-Tyr Chemical compound CC(C)[C@H](N)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](Cc1ccc(O)cc1)C(O)=O YLBNZCJFSVJDRJ-KJEVXHAQSA-N 0.000 description 1
- AOILQMZPNLUXCM-AVGNSLFASA-N Val-Val-Lys Chemical compound CC(C)[C@H](N)C(=O)N[C@@H](C(C)C)C(=O)N[C@H](C(O)=O)CCCCN AOILQMZPNLUXCM-AVGNSLFASA-N 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 235000004279 alanine Nutrition 0.000 description 1
- 108010044940 alanylglutamine Proteins 0.000 description 1
- 108010047495 alanylglycine Proteins 0.000 description 1
- 108010011559 alanylphenylalanine Proteins 0.000 description 1
- 239000002776 alpha toxin Substances 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 239000004411 aluminium Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000003466 anti-cipated effect Effects 0.000 description 1
- 108010008355 arginyl-glutamine Proteins 0.000 description 1
- 108010062796 arginyllysine Proteins 0.000 description 1
- 108010040443 aspartyl-aspartic acid Proteins 0.000 description 1
- 108010093581 aspartyl-proline Proteins 0.000 description 1
- 108010092854 aspartyllysine Proteins 0.000 description 1
- 108010068265 aspartyltyrosine Proteins 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 230000007969 cellular immunity Effects 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- OEYIOHPDSNJKLS-UHFFFAOYSA-N choline Chemical compound C[N+](C)(C)CCO OEYIOHPDSNJKLS-UHFFFAOYSA-N 0.000 description 1
- 229960001231 choline Drugs 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 229940125898 compound 5 Drugs 0.000 description 1
- 230000009514 concussion Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 238000005336 cracking Methods 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- UQLDLKMNUJERMK-UHFFFAOYSA-L di(octadecanoyloxy)lead Chemical compound [Pb+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O UQLDLKMNUJERMK-UHFFFAOYSA-L 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 239000000539 dimer Substances 0.000 description 1
- 238000005553 drilling Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 244000053095 fungal pathogen Species 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 108010078144 glutaminyl-glycine Proteins 0.000 description 1
- AWUCVROLDVIAJX-UHFFFAOYSA-N glycerol 1-phosphate Chemical compound OCC(O)COP(O)(O)=O AWUCVROLDVIAJX-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
- 108010019832 glycyl-asparaginyl-glycine Proteins 0.000 description 1
- 108010072405 glycyl-aspartyl-glycine Proteins 0.000 description 1
- 108010059898 glycyl-tyrosyl-lysine Proteins 0.000 description 1
- 108010020688 glycylhistidine Proteins 0.000 description 1
- 108010050848 glycylleucine Proteins 0.000 description 1
- 108010087823 glycyltyrosine Proteins 0.000 description 1
- 108010037850 glycylvaline Proteins 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000005802 health problem Effects 0.000 description 1
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 230000002209 hydrophobic effect Effects 0.000 description 1
- 230000028993 immune response Effects 0.000 description 1
- 230000003053 immunization Effects 0.000 description 1
- 238000002649 immunization Methods 0.000 description 1
- 230000006054 immunological memory Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 239000010985 leather Substances 0.000 description 1
- 108010044311 leucyl-glycyl-glycine Proteins 0.000 description 1
- 108010034529 leucyl-lysine Proteins 0.000 description 1
- 108010030617 leucyl-phenylalanyl-valine Proteins 0.000 description 1
- 108010073472 leucyl-prolyl-proline Proteins 0.000 description 1
- 108010057821 leucylproline Proteins 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 108010003700 lysyl aspartic acid Proteins 0.000 description 1
- 108010064235 lysylglycine Proteins 0.000 description 1
- 108010054155 lysyllysine Proteins 0.000 description 1
- 108010017391 lysylvaline Proteins 0.000 description 1
- 229910052749 magnesium Inorganic materials 0.000 description 1
- 239000011777 magnesium Substances 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 244000000010 microbial pathogen Species 0.000 description 1
- 230000002438 mitochondrial effect Effects 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 238000002703 mutagenesis Methods 0.000 description 1
- 231100000350 mutagenesis Toxicity 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 238000006386 neutralization reaction Methods 0.000 description 1
- -1 nitrite ions Chemical class 0.000 description 1
- 102000039446 nucleic acids Human genes 0.000 description 1
- 108020004707 nucleic acids Proteins 0.000 description 1
- 150000007523 nucleic acids Chemical class 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 108010079317 prolyl-tyrosine Proteins 0.000 description 1
- 239000012460 protein solution Substances 0.000 description 1
- 108010029895 rubimetide Proteins 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 108010069117 seryl-lysyl-aspartic acid Proteins 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 108010014657 sortilin Proteins 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 208000015339 staphylococcus aureus infection Diseases 0.000 description 1
- 229940030998 streptococcus agalactiae Drugs 0.000 description 1
- 239000013595 supernatant sample Substances 0.000 description 1
- 239000010414 supernatant solution Substances 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 108010017949 tyrosyl-glycyl-glycine Proteins 0.000 description 1
- 108010051110 tyrosyl-lysine Proteins 0.000 description 1
- 108010003137 tyrosyltyrosine Proteins 0.000 description 1
- 108010073969 valyllysine Proteins 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 239000001052 yellow pigment Substances 0.000 description 1
Classifications
-
- 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
- A61K39/02—Bacterial antigens
- A61K39/085—Staphylococcus
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K39/00—Medicinal preparations containing antigens or antibodies
- A61K2039/55—Medicinal preparations containing antigens or antibodies characterised by the host/recipient, e.g. newborn with maternal antibodies
- A61K2039/552—Veterinary vaccine
-
- 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
- A61K2039/55566—Emulsions, e.g. Freund's adjuvant, MF59
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Immunology (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
Abstract
The invention discloses a dairy cattle staphylococcus aureus mastitis subunit vaccine composition, a preparation method and application thereof. The composition comprises alpha-hemolysin protein, beta-hemolysin protein, PVL-S protein and pharmaceutically acceptable adjuvant or alpha-hemolysin protein, beta-hemolysin protein, PVL-F protein and pharmaceutically acceptable adjuvant; the preparation method comprises the following steps: 1) respectively preparing alpha-hemolysin protein, beta-hemolysin protein, PVL-F protein and PVL-S protein; 2) preparing antigen liquid from the alpha-hemolysin protein, the beta-hemolysin protein, the PVL-F protein or the alpha-hemolysin protein, the beta-hemolysin protein and the PVL-S protein prepared in the step 1); 3) preparing thimerosal according to the volume of the prepared composition, and adding the prepared thimerosal into the antigen solution in the step 2) to prepare a water phase; 4) the aqueous phase was emulsified by mixing with ISA 201 VG adjuvant in a volume ratio of 46: 55. The composition can induce organism to generate specific antibody, and generate good cross protection reaction.
Description
Technical field
The present invention relates to subunit vaccine composition of a kind of staphylococcus aureus mastitis in dairy cows and preparation method thereof
And application.Belong to live vaccine technical field.
Background technology
Mastitis for milk cows (Mastitis) is one of most common infectious diseases of adult dairy cattle, mainly cow mammary gland group
Knit by a kind of inflammation caused by after microorganism infection, multiple to be born in the postpartum breastfeeding phase, the disease is widely present in all over the world,
It is to cause one of Dairy Products Industry Implementing economic loss the most serious disease for milk cow common disease, frequently-occurring disease.Cause mastitis for milk cows
Pathogenic microorganism kind about more than 150, mainly based on staphylococcus aureus, Escherichia coli, Streptococcusagalactiae, these three are thin
Microbial mastitis for milk cows accounts for more than the 90% of total incidence, wherein especially using staphylococcus aureus as most.
The treatment of current mastitis for milk cows is mainly antibiotic therapy.Antibiotic is used to treat mastitis for milk cows existing more than 50
Year history, its served in the preventing and treating of mastitis for milk cows it is certain, but due to long-term single, heavy dose of, not science
Using antibiotic, sensitive bacteria elimination is caused, drug tolerant bacteria gradually account for main flow, especially staphylococcus aureus
Resistance problems are increasingly serious, result in antibiotic therapy and produce little effect；On the other hand, with the improvement of living standards, in milk
Antibiotic residue is a very serious health problem.
There is good prospect with vaccine control mastitis for milk cows, first, vaccine can prevent milk cow infection pathogen and
Cause mammitis；Secondly, vaccine helps to reduce the order of severity of intramammary infection, controls Subclinical mammitis；3rd, use
Vaccine control mammitis is not in antibiotic residue problem in milk；Be finally it is easy to operate, it is low-cost.Currently, develop
Successful vaccine is seldom, and most of is weak malicious live vaccine or inactivated vaccine, in production practices, has one to the preventing and treating of mammitis
It is set for using, however as the development of extensive intensive culture, the exquisite weak bacterial strain of people has homologous recombination, itself virulence and returned by force
Etc. potentially possible, and inactivated vaccine there is also dosage it is big, the deficiency such as not thorough is inactivated, to improve the security of traditional vaccine
With immune protective efficiency, efficient, cheap new generation vaccine develops particularly important.
Staphylococcus aureus (Staphylococcus aureus, abbreviation SA), is that a kind of leather is blue also referred to as " S. aureus L-forms "
The positive coccus of Albert'stain Albert, 0.8 μm of diameter is arranged in thyrsiform under the microscope, it is possible to produce golden yellow pigment, because
This and gain the name.S. aureus L-forms are to cause one of the main pathogenic fungi of chronic/recessive mastitis for milk cows, can be with strong influence milk
Yield and quality, huge economic loss is brought to dairy industry.
Alpha hemolysin (α-hemolysin), also known as α-toxin are a kind of exotoxins secreted by S. aureus L-forms, are that its is main
One of virulence factor, with good immunogenicity.Alpha hemolysin belongs to the beta-barrel structure bacteriotoxin family of hollow shape,
Relative molecular mass is 33,200, is made up of 297 amino acid, and its structural gene is hla.Alpha hemolysin is to a variety of mammals
Red blood cell has haemocylolysis, and its mechanism is the cell membrane hydrophobic region that lps molecule inserts red blood cell, forms micropore, destroys film
Integrality, cause cell to dissolve.By the hyte Histidine mutations of alpha hemolysin the 35th for after alanine, mutant just loses haemolysis
Activity, without toxicity, but still retains intact immunogenicity, can be used directly to immune animal, be S. aureus L-forms subunit epidemic disease
One of important candidate albumen of seedling.
β hemolysins (also known as β-toxin) are exactly one of its toxin, and it has the characteristics such as leucocytotoxicity, hemolytic activity.β
The single chain polypeptide that hemolysin is made up of 330 amino acid, molecular weight is 37~39kDa, and its isoelectric point (pI) is higher than 9.β haemolysis
Element is that a kind of magnesium relies on neural esterase, with phospholipase C (Phospholipase C, PLC) activity, can decompose phosphoglycerol
Choline, β hemolysins rely on this enzymatic activity, the phospholipid bilayer of hydrolysis composition cell membrane, so as to destroy the complete of cell membrane
Property, cause cell to crack, cause haemolysis.β hemolysins are carried out after rite-directed mutagenesis, mutant just loses hemolytic activity, does not have
It is toxic, but still retain intact immunogenicity, immune animal can be used directly to, is the important time of S. aureus L-forms subunit vaccine
One of sortilin.
Panton-valentine leukocidins (panton-valentine leukocidin, PVL) are by golden yellow
One of extracellular toxin that staphylococcus produces, is also one of its main virulence factor.PVL is by Van deleld in 1894
Find, and separated it from hemolysin by Panton and Valentine first in 1932.PVL is by two kinds of albumen
Matter is constituted, i.e. PVL-S albumen and PVL-F albumen, and its molecular weight is respectively between 34kDa and 33kDa, and F protein and S protein
Amino acid sequence homology has 36%.PVL belongs to film drilling toxin family, can induce PMNs necrosis or adjust and die, be PVL-S first
Combined with the acceptor of the specific high-affinity on PMNs cell membranes, secondly PVL-F formation dimers in combination, then successively
PVL-S and PVL-F knot platforms, eventually form the heteromers of a cyclic structure.This heteromers internal diameter is 3nm, and external diameter is 9nm, point
Son amount about 200kDa, PVL-S contained therein and PVL-F molecular proportion are 1:1, the heteromers of this cyclic structure are inserted in
On PMNs cell membranes, diameter about 2nm film perforation is formed, other PVL molecules enter cell by the hole, and online
Duct is set up on plastochondria outer membrane, so as to destroy mitochondrial interior environment, and caspase 9, caspase 3 and release is activated and kills white
Cytokine C, inducing cell, which is adjusted, dies.Therefore, PVL is one of important candidate albumen of S. aureus L-forms subunit vaccine, but works as PVL-S
With PVL-F all in the presence of, cytotoxicity is likely to result in, mouse safety experiment has been done in this laboratory to it, as a result with expection
It is identical, when the vaccine simultaneously containing PVL-S albumen and PVL-F albumen is immunized, mouse just death in 3-4 days after immune, still
Only it is immunized in the PVL-S albumen or PVL-F albumen of same dose, 14 days tracked after being immunized, mouse is all normal, does not go out incumbent
What is abnormal, so, in the selection of subunit vaccine, it is only capable of the candidate's egg for selecting one of which albumen as subunit vaccine
In vain.
This laboratory has separated more than 40 kinds of staphylococcus aureus from all parts of the country, and analysis has been carried out to its exotoxin grinds
Study carefully, one is to find that a kind of exotoxin is not only secreted in staphylococcus aureus strain of the same race, may secrete 2 kinds, 3 kinds or even more kinds of
Exotoxin；Two be that the exotoxin level that the staphylococcus aureus for finding that different regions are isolated secretes is not complete one
Sample, the mainly alpha hemolysin of some secretions, the mainly β hemolysins of some secretions.Therefore, using a kind of single exotoxin egg
It is white as subunit vaccine it is not only possible protect milk cow from staphylococcus aureus infection when effect it is not satisfactory, also not
The immune protective effect of wide spectrum can be provided.
The content of the invention
The technical problem to be solved in the present invention：One is to provide a kind of mastitis for milk cows caused by staphylococcus aureus
Subunit vaccine composition and preparation method thereof；Two be to overcome single ectotoxic subunit vaccine protecting milk cow from golden yellow
The problem of broad spectrum activity is poor in terms of color staphy lococcus infection；Three be to overcome existing Attenuate vaccine and inactivated vaccine that may be present homologous heavy
Group, itself virulence return the potential risk such as strong.
The invention provides a kind of staphylococcus aureus mastitis in dairy cows subunit vaccine composition, said composition is included
Alpha hemolysin albumen, beta hemolysin albumen, PVL-S albumen and pharmaceutically acceptable adjuvant, or alpha hemolysin albumen, β-molten
Sanguinin albumen, PVL-F albumen and pharmaceutically acceptable adjuvant.
In technical scheme, it is preferable that the encoding gene of the PVL-S albumen is as shown in SEQ ID NO.1.
In technical scheme, it is preferable that the encoding gene of the PVL-F albumen is as shown in SEQ ID NO.2.
In technical scheme, it is preferable that the alpha hemolysin albumen be it is treated forfeiture hemolytic activity but
Retain the albumen of immunogenicity, the processing method has albumen inactivation and gene mutation processing.Preferably, the alpha hemolysin egg
It is the albumen lost hemolytic activity but retain immunogenicity handled by gene mutation in vain.
In technical scheme, it is preferable that the beta hemolysin albumen be it is treated forfeiture hemolytic activity but
Retain the albumen of immunogenicity, the processing method has albumen inactivation and gene mutation processing.Preferably, the beta hemolysin egg
It is the albumen lost hemolytic activity but retain immunogenicity by mutation processing in vain.
In technical scheme, it is preferable that the alpha hemolysin albumen, beta hemolysin albumen, PVL-S albumen according to
Deng mass ratio mixing.
In technical scheme, it is preferable that the alpha hemolysin albumen, beta hemolysin albumen, PVL-F albumen according to
Deng mass ratio mixing.
In technical scheme, it is preferable that the alpha hemolysin albumen, beta hemolysin albumen, PVL-S albumen are
100 parts of μ g/.
In technical scheme, it is preferable that the alpha hemolysin albumen, beta hemolysin albumen, PVL-F albumen are
100 parts of μ g/.
In technical scheme, it is preferable that the pharmaceutically acceptable adjuvant can be aqueous adjuvants (such as aluminium glue
Adjuvant etc.), or oil-in-water adjuvant (such as white oil), or W/O/W adjuvant (such as ISA 206V), it is excellent
Selection of land, the pharmaceutically acceptable adjuvant is ISA201VG adjuvants.
In technical scheme, it is preferable that the composition also contains preservative.
In technical scheme, it is preferable that described preservative is thimerosal, and the content of the thimerosal is 2 μ g/
Head part.
Present invention also offers a kind of method for preparing the subunit vaccine composition, it the described method comprises the following steps：
1) alpha hemolysin albumen, beta hemolysin albumen, PVL-F albumen, PVL-S albumen are prepared respectively；2) by step 1) according to etc. matter
Amount than the alpha hemolysin albumen, beta hemolysin albumen, the PVL-F albumen that are mixed with, or according to etc. mass ratio be mixed with α-
Haemolysis fibroin, beta hemolysin albumen, PVL-S albumen are prepared into composition antigen liquid；3) according to step 2) the middle combination prepared
The volume of thing antigen liquid, gets out thimerosal, and ready thimerosal is added into step 2) aqueous phase is prepared into antigen liquid；
4) by the aqueous phase and ISA 201VG adjuvants according to volume ratio 46:55 mixing and emulsifyings.Preferably, step 4) described in emulsify
Temperature is 32-35 DEG C.
The present invention provides a kind of composition and prepared for preventing and treating milk cow staphylococcus aureus breast again
Application in the scorching vaccine in room.
Compared with prior art, the present invention clearly provides a kind of new milk cow staphylococcus aureus breast for the first time
Scorching subunit vaccine composition and its preparation method and application.Said composition does not contain nucleic acid first, will not cause homologous recombination
The potential danger such as strong is returned with itself virulence；Body can be induced and produce specific antibody, cellular immunity, inducing immunological memory can be promoted
With cause extensive immune response, produce good cross protection reaction, thus the vaccine more safety and stability, prepare it is simple,
Using convenient, cheap and time saving and energy saving.Secondly, said composition is due to containing a variety of staphylotoxins, therefore
Different ectotoxic staphylococcus aureuses, which can be secreted, to different regions good neutralization, reaches that protection milk cow is exempted from
Infected by different ectotoxic staphylococcus aureuses are secreted；Finally, said composition is than subunit prepared by single exotoxin
Vaccine is protecting milk cow stronger from the ability in terms of infection of staphylococcus aureus.
Brief description of the drawings
Fig. 1 represents that agarose gel electrophoresis PCR expands PVL-F, PVL-S results.PVL-F gene PCR results, size is about
906bp；PVL-S gene PCR results, size is about 849bp；M：DNA molecular amount standard DL2,000.
Fig. 2 represents pET28a-PVL-F, PVL-S digestion verification results.pET28a-PVL-F：Plasmid pET28a-PVL-F
XhoI and Nde I digestion qualification results, endonuclease bamhi size is 5,369bp and 904bp；pET28a-PVL-S：Plasmid pET28a-
PVL-F XhoI and Nde I digestion qualification results, endonuclease bamhi size is 5,369bp and 849bp；M：DNA molecular amount standard
DL5,000。
Fig. 3 represents PVL-F albumen and PVL-S protein purification results.
Fig. 4 a represent the bioactivity result of alpha hemolysin in immune rear composition；
Fig. 4 b represent the bioactivity result of beta hemolysin in immune rear composition；
Fig. 4 c represent the bioactivity result of PVL-F in immune rear composition；
Fig. 4 d represent the bioactivity result of PVL-S in immune rear composition.
Challenge viral dosage result after Fig. 5 is immune.
Embodiment
Below with reference to drawings and examples, the present invention will be further described, and embodiments of the invention are merely to illustrate this
The technical scheme of invention, and the non-limiting present invention.
The source list of reagent and medicine of the present invention is as follows：
Chemical reagent and all commercially available prod of biological reagent；
Preservative thimerosal is purchased from Life Sciences；
ISA201VG is purchased from match BIC Corp of France.
Embodiment 1：It is prepared by alpha hemolysin albumen, beta hemolysin albumen, PVL-S albumen, PVL-F albumen
1.1：It is prepared by alpha hemolysin albumen
The patent of invention for the Patent No. 201610068723.4 submitted with reference to the applicant prepares the alpha hemolysin of mutation
Albumen.
1.2：It is prepared by beta hemolysin albumen
The patent of invention for the Patent No. 201610068816.7 submitted with reference to the applicant prepares the beta hemolysin of mutation
Albumen.
1.3：It is prepared by PVL-S albumen, PVL-F albumen
1.3.1 expression vector pET28a-PVL-S and pET28a-PVL-F structure (two kinds of carrier construction methods are identical)
Milk cow staphylococcus aureus gene group to be clinically separated enters performing PCR as template, and primer is as shown in the table,
As a result it is as shown in Figure 1：PVL-F gene PCR results, size is about 906bp；PVL-S gene PCR results, size is about
849bp；It is all in the same size with expection.
1.3.1.1 sample-adding system is (50 μ l):
1.3.1.2PCR amplification program:
1.3.1.3 glue reclaim DNA fragmentation:
(1) step 1.3.1.2 reaction solution is subjected to 0.8% agarose gel electrophoresis (110V 30min)；
(2) under uviol lamp, gel extraction DNA fragmentation is in 1.5ml EP pipes；
(3) 500 μ l PC buffer, 50 DEG C, water-bath 10min are added in the 1.5ml EP pipes into step (2)；
(4) solution in step (3) is moved into adsorption column center, stands 2min, centrifugation, 12,000rpm, 30s；
(5) waste liquid is abandoned, 600 μ l PW buffer are added to adsorption column center, 3min, 12,000rpm, 30s is stood；
(6) repeat step (5)；
(7) suction attached column is centrifuged, 12,000rpm, 1min；
(8) 30 μ l ddH are added to adsorption column center2O, stands 3min, centrifuges (12,000rpm, 2min)；
(9) collection step (8) DNA sample carries out electrophoresis.
1.3.1.4 double digestion reaction (50 μ l systems)：
It is loaded, mixed according to above-mentioned system in 1.5ml EP pipes, the two 50 μ l reaction solutions is then placed in 37
In DEG C thermostat water bath, water-bath 3h.
1.3.1.5 glue reclaim DNA fragmentation：
(1) step 1.3.1.4 reaction solution is subjected to 0.8% agarose gel electrophoresis (110V 30min)；
(2) under uviol lamp, gel extraction DNA fragmentation is in 1.5ml EP pipes；
(3) 500 μ l PC buffer, 50 DEG C, water-bath 10min are added in the 1.5ml EP pipes into step (2)；
(4) solution in step (3) is moved into adsorption column center, stands 2min, centrifugation, 12,000rpm, 30s；
(5) waste liquid is abandoned, 600 μ l PW buffer are added to adsorption column center, 3min, 12,000rpm, 30s is stood；
(6) repeat step (5)；
(7) suction attached column is centrifuged, 12,000rpm, 1min；
(8) 30 μ l ddH are added to adsorption column center2O, stands 3min, centrifuges (12,000rpm, 2min)；
(9) collection step (8) DNA sample carries out electrophoresis.
1.3.1.6 coupled reaction (10 μ l systems)：
It is loaded, mixed according to above-mentioned system in 1.5ml EP pipes, above-mentioned reaction solution is then placed in 16 DEG C, water-bath
Take out, 65 DEG C, inactivated after water-bath 15min after 16h, by 4 DEG C of preservations of sample.
1.3.1.7 transformation experiment：
(1) step 1.3.1.6 coupled reaction liquid is taken out, 100 μ l E.coli DH5 α competent cells are added thereto,
Mix；
(2) ice bath 30min；
(3) 42 DEG C, water-bath 100s；
(4) ice bath 2min；
(5) take out, 600 μ l LB liquid mediums, 37 DEG C, water-bath 1h are added into EP pipes；
(6) sample cell is taken out, is centrifuged (8,000rpm, 2min), removes 600 μ l, remaining 100 μ l LB and thalline is resuspended；
(7) take bacterium solution to be plated in LK flat boards (Kan concentration is 50 μ g/ml), LK flat boards are placed in biochemical constant incubator
In, 37 DEG C of culture 12h.
1.3.1.8 recombinant plasmid is extracted and digestion identification：
(1) picking monoclonal is into 3ml LK fluid nutrient mediums from conversion flat board, and 37 DEG C, 260rpm shakes bacterium and stayed overnight；
(2) 1ml bacterium solutions are taken into 1.5ml EP pipes, centrifuges (12,000rpm, 2min), abandons supernatant；
(3) 250 μ l P1buffer are added in the EP pipes into step (2), thalline is resuspended；
(4) 250 μ l P2buffer are added into step (3) solution, it is gentle to mix, stand 2min；
(5) 350 μ l P3buffer are added into step (4) solution, it is gentle to mix；
(6) by step (5) solution, centrifuge (12,000rpm, 10min)；
(7) supernatant solution in step (6) is moved into adsorption column center, centrifuged (8,000g, 30s)；
(8) waste liquid is abandoned, 500 μ l wash buffer are added to adsorption column center, is centrifuged (9,000g, 30s)；
(9) repeat step (8)；
(10) suction attached column centrifugation (9,000g, 1min)；
(11) 30 μ l Elution buffer are added to adsorption column, stands 2min, centrifuged (12,000rpm, 2min)；
(12) collection step (11) DNA sample carries out electrophoresis；
(13) as shown in step 1.3.1.4, digestion identification is carried out to the plasmid extracted, 0.8% agarose is then carried out
Gel electrophoresis.
(14) recombinant plasmid digestion qualification result is as shown in Figure 2：pET28a-PVL-F：Plasmid pET28a-PVL-F XhoI
With Nde I digestion qualification results, endonuclease bamhi size is 5,369bp and 904bp；pET28a-PVL-S：Plasmid pET28a-PVL-
F XhoI and Nde I digestion qualification results, endonuclease bamhi size is 5,369bp and 849bp.
1.3.2 e. coli bl21 is converted
Draw 1 μ l plasmids to add in 100 μ l BL21 competent cells, ice bath 30min；
42 DEG C of heat shock 90s；
Ice bath 2min；
The LB nutrient solutions of 900 μ l non-resistants are added in super-clean bench；
37 DEG C of 180rpm shake 1h；
Draw 100 μ l bacterium solutions card-coating that resistance LB flat boards, 37 DEG C of incubated overnights.
1.3.3 a large amount of induced expressions
Choose bacterium：Picking monoclonal is into 50ml cards that resistance LB nutrient solutions, 37 DEG C of incubated overnights；
Switching：By 1:100 ratios transfer bacterium solution to 500ml cards that resistance LB nutrient solutions, and 3.5L, 37 DEG C of 220rpm trainings are shaken altogether
Support 2-2.5h to OD600It is worth 0.6；
Induction：Bacterium solution OD600After being worth 0.6,500 μ l IPTG (1M) are added to the final concentration of 1mmol/L of IPTG, 37 DEG C
220rpm Fiber differentiations 4h；
Microorganism collection：Bacterium solution 6,000rpm centrifugation 10min, collects thalline；With 40ml PBS thalline, 6,000rpm from
Heart 10min, collects thalline, is placed in -20 DEG C of preservations；
1.3.4PVL-S albumen or PVL-F protein purifications (two kinds of protein purification procedures are identical)
(1) bacterial cell disruption：Lysate (8ml/g weight in wet bases) (50mM NaH are added in obtained thalline into 1.3.32PO4
(pH 8.0), 500mM NaCl, 20mM imidazoles；Membrane filtration using 0.8 μm), blown and beaten with 50mL syringes uniform to nothing
Granular fungus block；Thalline sample is poured into cell homogeneous instrument sample cell, outlet is prepared to collect sample with beaker；With sample
90% outflow outlet is a circulation, and after the completion of a circulation, the sample that outlet beaker is collected refunds sample cell, continues
Repeat 5 circulations.
(2) complete sample will be crushed in step (1) to be dispensed into 250mL Beckman centrifuge tubes, 12,000rpm, 4 DEG C
30min is centrifuged, Supernatant samples are crossed after 0.8 μm of film as loading sample, reserve 80 μ L samples and detected for SDS-PAGE；
(3) column equilibration：With the ultrapure column volume of water balance 2~3 (CV), the ethanol of discharge 20% preserves liquid；Then with cracking
Liquid balances 2~3 column volumes (CV), 5mL/min.Control pressure is less than 0.5MPa.
(4) loading：2mL/min carries out loading, and collection flows through liquid (Flowthrough), takes 80 μ L to be examined for SDS-PAGE
Survey.
(5) 1 is rinsed：With (the washing buffer 1 of cleaning buffer solution component 1：50mM NaH2PO4(pH 7.4), 500mM
NaCl, 20mM imidazoles, 0.1%TritonX-114, the membrane filtration using 0.8 μm) post is washed, 5mL/min rinses 80 cylinders
Product.
(6) 2 are rinsed：With (the elution buffer1 of elution buffer component 1 of 10 times of column volumes (CV)：50mM
NaH2PO4(pH 7.4), 500mM NaCl, the membrane filtration using 0.8 μm) post is washed, reduce Triton X-114 residual.
(7) elute：(the 50%Elution buffer 2 of 50% elution buffer component 2：50mM NaH2PO4(pH 7.4),
500mM NaCl, 250mM imidazoles, the membrane filtration using 0.8 μm) elution destination protein, wash flat to baseline, speed 5ml/min,
Collect, take 80 μ L to be analyzed for SDS-PAGE after mixing；The 100% de- component of buffer solution 2 cleaning pillar (100%Elution
buffer 2：50mM NaH2PO4(pH 7.4), 500mM NaCl, 500mM imidazoles, the membrane filtration using 0.8 μm), to baseline
Wash flat, 5mL/min is collected, and takes 80 μ L to be analyzed for SDS-PAGE after mixing.
(8) HiPrep Desalting desalting columns column equilibration：With the ultrapure column volume of water balance 2~3 (CV), 20% is discharged
Ethanol preserves liquid；Then 3-4 column volumes (CV), speed 10mL/min are balanced with elution buffer component 1.
(9) loading：Speed 10mL/min is injected (inject), and maximum injection (inject) amount is 13mL.
(10) collect：Stop after loading, into load patterns, 10mL/min, UV rises to 1mAU and starts collection, i.e. albumen
Sample starts appearance, and 10ml/ pipes are collected, and treats that UV is down to below 5mAU, stops collecting.
(11) balance：Flow velocity 10ml/min, balances 2-3CV.
(12) 7.10~7.12 are circulated, until completion of the sample.
(13) aseptic filtration：The protein solution of collection is at 4 DEG C, and 12,000rpm centrifugation 15min collect supernatant, move to biology
Safety cabinet, crosses the low syringe needle filter of 0.2 μm of protein binding rate, and -80 DEG C of refrigerator preservations are placed in after filtering.
(14) purification result is as shown in Figure 3：PVL-S albumen and the purity of PVL-F albumen after purification can reach 90%
More than.
Embodiment 2：Composition is prepared and (illustrated exemplified by preparing 1ml/ parts)
The consumptive material and material used for preparing vaccine all need it is pre- first pass through aseptic process, preparation process be in Biohazard Safety Equipment or
Other can ensure to complete in whole preparation process all sterile instrument or environment.
(1) needed according to experiment, the amount of each composition of calculation composition solution, make alpha hemolysin albumen, β in composition-molten
Sanguinin albumen, PVL-S albumen or alpha hemolysin albumen, beta hemolysin albumen, PVL-F final concentration of protein are 100 μ g/ml, are made
The final concentration of 2 μ g/ml of thimerosal in composition, then load weighted recombinant protein is mixed with load weighted thimerosal be used as water
Phase, is 46 according to aqueous phase and adjuvant ISA201VG volume ratios:55, measure adjuvant；
(2) measured aqueous phase and adjuvant are placed in thermostat water bath and are heated to 33 DEG C ± 1 DEG C, after general after temperature stabilization
Antigen is added in adjuvant pipe, and oscillator concussion 10min carries out pre-emulsification；
(3) the complete vaccine of pre-emulsification is positioned in the beaker for filling with ice, is fixed on the supersonic cell anticipated
Emulsified on broken instrument；
(4) after emulsification terminates, emulsifying effectiveness is observed：Part vaccine is taken to be placed in centrifuge tube, 3,000rpm centrifugation 15min,
Vaccine is not stratified to be qualified；
(5) detect that qualified vaccine is dispensed into 15ml centrifuge tubes, mark that sealed membrane sealing is placed in 4 DEG C of preservations.
Embodiment 3：Mouse immune challenge viral dosage
3.1：Method according to embodiment 2 prepares following vaccine and composition
3.2：Immunization experiment
80 20g of purchase or so Balb/c female mices, are classified as 8 groups, every group 10, one of which is used as control
The immune PBS of group, 7 groups as immune group in addition, and the 7 kinds of compositions prepared in 3.1 are immunized respectively；Inhaled when immune with 1ml syringes
1ml vaccines are taken, are then sterilized with 75% cotton ball soaked in alcohol to mouse hind leg muscle, the inserting needle in the middle part of muscle masses, left and right back leg is respectively noted
Penetrate 50 μ l vaccines, the μ l vaccines of co-injection 100.
Two exempt within 14 days after one exempting from, two exempt from after carry out within 7 days three and exempt from, and before exempting from one, two exempt from before, three exempt from before and three exempt from
14 days collection serum, detects antibody titer afterwards.
Bioactivity result is as shown in Fig. 4 a, Fig. 4 b, Fig. 4 c, Fig. 4 d, wherein the composition (composition containing alpha hemolysin
1st, composition 5, composition 6, composition 7) in the relative potency of alpha hemolysin 544, more than 000 can be reached before poison is attacked,
It is 544,000, the relative potency of other 3 compositions reaches but the relative potency of composition 6 is minimum in 4 compositions
More than 928,000, this (high 10%) of survival rate of the survival rate of composition 7 than composition 6 also consistent with attacking malicious result；Containing β-
The relative potency of beta hemolysin in the composition (composition 2, composition 5, composition 6, composition 7) of hemolysin is before poison is attacked
More than 77,778 can be reached；PVL-F's in composition (composition 3, composition 6, composition 7) containing PVL-F is relative
Potency can reach more than 80,000 before poison is attacked；In composition (composition 4, composition 6, composition 7) containing PVL-S
PVL-S relative potency can reach more than 80,000 before poison is attacked.
3:3：Challenge viral dosage (three exempt to carry out within latter 14 days attacking poison)
(1) picking SA single bacterium colonies (bacterial strain is the CQ339 bacterial strains that this laboratory is preserved) are in 5ml liquid broths
In, 220rpm, 37 DEG C shake bacterium and stay overnight；
(2) microbionation overnight will be shaken in 100ml fresh liquid broth bouillons by centesimal volume (1ml)
In, 220rpm, 37 DEG C shake bacterium and stay overnight；
(3) 100ml bacterium solutions are encased in 500ml centrifugal bottles, 8,000rpm centrifugation 10min suck culture medium, thalline is used
100ml PBS are resuspended, and repeat the above steps 3 times, and finally all thalline are resuspended with 5ml PBS and mix；
(4) counted after bacterium solution being done into 10,000 times of dilutions.According to the concentration of bacterium solution, original bacteria liquid is diluted to 1.5 ×
108CFU/ml(2M LD50)；
(5) bacterium solution is drawn with 1ml syringes, tail vein injection, injection volume is carried out according to the corresponding mouse of each concentration bacterium
For 200 μ l/20g mouse.
(6) survival state of Continuous Observation mouse, records the death time of each mouse：It is each in the morning, afternoon and evening to check once.
(7) mouse survival rate result after poison is attacked as shown in Fig. 2 attacking 500h internal references group mouse all death after poison；With
Within the 987.5h of track, 7 groups of immune group survival rates are all higher than 50%, wherein, the survival rate of composition 1- compositions 4 exists
50%-60%, high 20%-30% of the survival rate than composition 1- compositions 4 of composition 5, reach 70%, and composition 6 is deposited
Motility rate reaches 80%, and the survival rate of composition 7 reaches 90%, all the significantly larger than survival rate of composition 1- compositions 4, also than group
The survival rate of compound 5 will height, the survival rate of composition 7 is higher than the survival rate of composition 6 by 10%, this and alpha hemolysin in composition
Bioactivity result is consistent, and (high one of the potency of the alpha hemolysin in alpha hemolysin potency ratio composition 6 in composition 7 is dilute
Release ratio)；The immune protective effect that the exotoxin of this explanation one-component prepares vaccine can not show a candle to a variety of exotoxins and prepare vaccine
Immune protective effect.
Embodiment 4：ELISA antibody titers are detected
(1) it is coated with：Detection albumen (the alpha hemolysis of purifying is diluted with coating buffer (50mM carbonate buffer solutions, pH 9.5)
Fibroin, beta hemolysin albumen, PVL-S albumen or PVL-F albumen) to 0.5 μ g/ml, 100 μ l are added per hole on ELISA Plate,
4 DEG C of refrigerators are stood overnight after sealed membrane is sealed；
(2) wash：Taken out from refrigerator after ELISA Plate, be put into board-washing machine and wash, cleaning solution PBST；
(3) close：200 μ l confining liquids (5% defatted milk), 37 DEG C of incubation 2h after sealed membrane is sealed are added per hole；
(4) preparation of samples：By known information and consumption is needed, serum is subjected to appropriateness dilution with confining liquid；
(5) wash：With (2)；
(6) it is loaded：Dilute serum is added, while negative control is made of confining liquid, 37 DEG C of incubation 1h；
(7) wash：With (2)；
(8) secondary antibody is added：Secondary antibody 100 the μ l, 37 DEG C of incubation 0.5h of the HRP marks of appropriateness dilution are added per hole；
(9) wash：With (2)；
(10) develop the color：100 μ l TMB nitrite ions, 37 DEG C of incubation 10min are added under the conditions of lucifuge per hole；
(11) terminate：50 μ l terminate liquids (2M H are added per hole2SO4), terminating reaction；
(12) detect：In 450nm wavelength determination sample OD values, analyze data；
(13) interpretation of result：Judge the standard of antibody positive：P/N >=2.1, OD450 >=0.1.
The present invention is illustrated by above embodiment, it is understood, however, that the present invention is not limited to institute here
The particular example and embodiment of description.Purpose herein comprising these particular examples and embodiment is to help this area
In technical staff practice the present invention.Any those of skill in the art are easy to do not departing from spirit and scope of the invention
In the case of be further improved and perfect, therefore the present invention only by the content of the claims in the present invention and limiting for scope
System, its intention covers the alternative in all spirit and scope of the invention for being included in and being limited by appendix claim and waited
Same scheme.
Sequence table
<110>Zhejiang oceanic rise bio tech ltd
<120>Staphylococcus aureus mastitis in dairy cows subunit vaccine and its preparation method and application
<160> 4
<170> PatentIn version 3.3
<210> 4
<211> 879
<212> DNA
<213>Recombination staphylococcus aureus PVL-S protein coding gene sequences
<400> 1
atgggaaata ttgagaatat tggtgatggc gctgaggtag tcaaaagaac agaagataca 60
agtagcgata agtggggggt cacacaaaat attcagtttg attttgttaa agataaaaag 120
tataacaaag acgctttgat tttaaaaatg caaggtttta tcaattcaaa gactacttat 180
tacaattaca aaaacacaga tcatataaaa gcaatgaggt ggcctttcca atacaatatt 240
ggtctcaaaa caaatgaccc caatgtagat ttaataaatt atctacctaa aaataaaata 300
gattcagtaa atgttagtca aacattaggt tataacatag gtggtaattt taatagtggt 360
ccatcaacag gaggtaatgg ttcatttaat tattcaaaaa caattagtta taatcaacaa 420
aactatatca gtgaagtaga acgtcaaaat tcaaaaagtg ttcaatgggg aataaaagct 480
aattcattta tcacatcatt aggtaaaatg tctggacatg atccaaattt atttgttgga 540
tataaaccat atagtcaaaa tccgagagac tattttgttc cagacaatga attaccccca 600
ttagtacaca gtggtttcaa tccttcattt attgcaactg tttctcatga aaaaggctca 660
ggagatacaa gtgaatttga aataacgtat ggcagaaata tggatgttac tcatgctact 720
agaagaacaa cacactatgg caatagttat ttagaaggat ctagaataca caacgcattt 780
gtaaacagaa attacacagt taaatatgaa gtgaactgga aaactcatga aattaaagtg 840
aaaggacata atctcgagca ccaccaccac caccactga 879
<210> 4
<211> 921
<212> DNA
<213>Recombination staphylococcus aureus PVL-F protein coding gene sequences
<400> 2
atgggagctc aacatatcac acctgtcagc gagaaaaaag tggatgacaa aatcactttg 60
tacaaaacga ctgctacatc agattctgac aaattaaaaa tttctcaaat tctaactttt 120
aattttatta aagacaaaag ttatgataaa gacacattaa tactaaaagc tgccggaaac 180
atttactcag gctataccca acccacttct gatagtagta taaattcaca attttattgg 240
ggagctaagt ataatgtttt tgttagctcg gagtccaaag attctgtaaa tattgttgac 300
tacgcgccta aaaatcaaaa tgaagaattt caagttcaac aaacattagg ttattcatat 360
ggcggagata ttaatataat aaatggatta actggtggat tgaacgggtc aaaatcattt 420
tcagaaacga ttaattataa gcaagaaagc tacagaacta cgattgatag gaaaataaat 480
cacaaattaa tcggctgggg tgtcgaggca cataaaatca tgaataatgg ttggggacca 540
tatggcagag atagtagtga ttcattatat ggaaacgaac tatttttagg tggcagacag 600
agtagctcga atgctaatca aaatttctta ccaacacatc aaatgcccat attagcacgt 660
ggtaatttca atccagaatt tataagcgta ctttctcaca aacaaaagga tgttaaaaaa 720
tctaaaatta aagtgactta tcaaagacaa atggatcggt atgaaaattt ttggaacaac 780
ttgcactgga taggttataa tattaagaat caaaagagag caacacacac atcaatttat 840
gaaattgatt gggaaaaaca cacggttaaa ttagtagctt cgcaatctag cgaactcgag 900
caccaccacc accaccactg a 921
<210> 1
<211> 292
<212> PRT
<213>Recombination staphylococcus aureus PVL-S protein sequences
<400> 3
Met Gly Asn Ile Glu Asn Ile Gly Asp Gly Ala Glu Val Val Lys Arg
1 5 10 15
Thr Glu Asp Thr Ser Ser Asp Lys Trp Gly Val Thr Gln Asn Ile Gln
20 25 30
Phe Asp Phe Val Lys Asp Lys Lys Tyr Asn Lys Asp Ala Leu Ile Leu
35 40 45
Lys Met Gln Gly Phe Ile Asn Ser Lys Thr Thr Tyr Tyr Asn Tyr Lys
50 55 60
Asn Thr Asp His Ile Lys Ala Met Arg Trp Pro Phe Gln Tyr Asn Ile
65 70 75 80
Gly Leu Lys Thr Asn Asp Pro Asn Val Asp Leu Ile Asn Tyr Leu Pro
85 90 95
Lys Asn Lys Ile Asp Ser Val Asn Val Ser Gln Thr Leu Gly Tyr Asn
100 105 110
Ile Gly Gly Asn Phe Asn Ser Gly Pro Ser Thr Gly Gly Asn Gly Ser
115 120 125
Phe Asn Tyr Ser Lys Thr Ile Ser Tyr Asn Gln Gln Asn Tyr Ile Ser
130 135 140
Glu Val Glu Arg Gln Asn Ser Lys Ser Val Gln Trp Gly Ile Lys Ala
145 150 155 160
Asn Ser Phe Ile Thr Ser Leu Gly Lys Met Ser Gly His Asp Pro Asn
165 170 175
Leu Phe Val Gly Tyr Lys Pro Tyr Ser Gln Asn Pro Arg Asp Tyr Phe
180 185 190
Val Pro Asp Asn Glu Leu Pro Pro Leu Val His Ser Gly Phe Asn Pro
195 200 205
Ser Phe Ile Ala Thr Val Ser His Glu Lys Gly Ser Gly Asp Thr Ser
210 215 220
Glu Phe Glu Ile Thr Tyr Gly Arg Asn Met Asp Val Thr His Ala Thr
225 230 235 240
Arg Arg Thr Thr His Tyr Gly Asn Ser Tyr Leu Glu Gly Ser Arg Ile
245 250 255
His Asn Ala Phe Val Asn Arg Asn Tyr Thr Val Lys Tyr Glu Val Asn
260 265 270
Trp Lys Thr His Glu Ile Lys Val Lys Gly His Asn Leu Glu His His
275 280 285
His His His His
290
<210> 1
<211> 306
<212> PRT
<213>Recombination staphylococcus aureus PVL-F protein sequences
<400> 4
Met Gly Ala Gln His Ile Thr Pro Val Ser Glu Lys Lys Val Asp Asp
1 5 10 15
Lys Ile Thr Leu Tyr Lys Thr Thr Ala Thr Ser Asp Ser Asp Lys Leu
20 25 30
Lys Ile Ser Gln Ile Leu Thr Phe Asn Phe Ile Lys Asp Lys Ser Tyr Asp
35 40 45
Lys Asp Thr Leu Ile Leu Lys Ala Ala Gly Asn Ile Tyr Ser Gly Tyr
50 55 60 65
Thr Gln Pro Thr Ser Asp Ser Ser Ile Asn Ser Gln Phe Tyr Trp Gly
70 75 80
Ala Lys Tyr Asn Val Phe Val Ser Ser Glu Ser Lys Asp Ser Val Asn
85 90 95
Ile Val Asp Tyr Ala Pro Lys Asn Gln Asn Glu Glu Phe Gln Val Gln
100 105 110
Gln Thr Leu Gly Tyr Ser Tyr Gly Gly Asp Ile Asn Ile Ile Asn Gly
115 120 125
Leu Thr Gly Gly Leu Asn Gly Ser Lys Ser Phe Ser Glu Thr Ile Asn
130 135 140 145
Tyr Lys Gln Glu Ser Tyr Arg Thr Thr Ile Asp Arg Lys Ile Asn His
150 155 160
Lys Leu Ile Gly Trp Gly Val Glu Ala His Lys Ile Met Asn Asn Gly
165 170 175
Trp Gly Pro Tyr Gly Arg Asp Ser Ser Asp Ser Leu Tyr Gly Asn Glu
180 185 190
Leu Phe Leu Gly Gly Arg Gln Ser Ser Ser Asn Ala Asn Gln Asn Phe
195 200 205
Leu Pro Thr His Gln Met Pro Ile Leu Ala Arg Gly Asn Phe Asn Pro
210 215 220 225
Glu Phe Ile Ser Val Leu Ser His Lys Gln Lys Asp Val Lys Lys Ser
230 235 240
Lys Ile Lys Val Thr Tyr Gln Arg Gln Met Asp Arg Tyr Glu Asn Phe
245 250 255
Trp Asn Asn Leu His Trp Ile Gly Tyr Asn Ile Lys Asn Gln Lys Arg
260 265 270
Ala Thr His Thr Ser Ile Tyr Glu Ile Asp Trp Glu Lys His Thr Val Lys
275 280 285 290
Leu Val Ala Ser Gln Ser Ser Glu Leu Glu His His His His His His
295 300 305
Claims (13)
1. a kind of composition of staphylococcus aureus mastitis in dairy cows subunit vaccine, it is characterised in that：The composition bag
Albumen containing alpha hemolysin, beta hemolysin albumen, PVL-S albumen and pharmaceutically acceptable adjuvant, or alpha hemolysin albumen, β-
Haemolysis fibroin, PVL-F albumen and pharmaceutically acceptable adjuvant.
2. composition according to claim 1, it is characterised in that the alpha hemolysin albumen is handled by gene mutation
Lose hemolytic activity but retain immunogenicity albumen.
3. composition according to claim 1, it is characterised in that the beta hemolysin albumen is handled by gene mutation
Lose hemolytic activity but retain immunogenicity albumen.
4. composition according to claim 1, it is characterised in that the alpha hemolysin albumen, beta hemolysin albumen, PVL-
S protein is according to grade mass ratio mixing.
5. composition according to claim 1, it is characterised in that the alpha hemolysin albumen, beta hemolysin albumen, PVL-
F protein is according to grade mass ratio mixing.
6. the composition according to claim 1 or 4, it is characterised in that the alpha hemolysin albumen, beta hemolysin albumen,
PVL-S albumen is g/ parts of 100 μ.
7. composition according to claim 1 or 5, it is characterised in that the alpha hemolysin albumen, beta hemolysin albumen,
PVL-F albumen is g/ parts of 100 μ.
8. composition according to claim 1, it is characterised in that the pharmaceutically acceptable adjuvant is the VG of ISA 201
Adjuvant.
9. the composition according to claim 1 to 8 any claim, it is characterised in that the composition is also containing anti-
Rotten agent.
10. composition according to claim 9, it is characterised in that the preservative is thimerosal.
11. composition according to claim 10, it is characterised in that the content of the thimerosal is g/ parts of 2 μ.
12. a kind of method for preparing the composition as described in claim 1 to 11 is any, it is characterised in that methods described includes
Following steps：
1) alpha hemolysin albumen, beta hemolysin albumen, PVL-F albumen, PVL-S albumen are prepared respectively；
2) by step 1) according to waiting mass ratio the alpha hemolysin albumen, beta hemolysin albumen, the PVL-F albumen that are mixed with, or
Alpha hemolysin albumen, beta hemolysin albumen, PVL-S albumen according to waiting mass ratio to be mixed with are prepared into composition antigen liquid；
3) according to step 2) in prepare composition antigen liquid volume, get out thimerosal, and ready thimerosal is added
Enter to step 2) aqueous phase is prepared into antigen liquid；
4) by the aqueous phase and the VG adjuvants of ISA 201 according to volume ratio 46:55 mixing and emulsifyings.
13. a kind of composition as described in claim 1-11 any claims is golden yellow for preventing and treating milk cow in preparation
Application in the vaccine of color aureus mastitis.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710128121.8A CN107224575B (en) | 2017-03-06 | 2017-03-06 | Composition of dairy cow staphylococcus aureus mastitis subunit vaccine and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710128121.8A CN107224575B (en) | 2017-03-06 | 2017-03-06 | Composition of dairy cow staphylococcus aureus mastitis subunit vaccine and preparation method and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107224575A true CN107224575A (en) | 2017-10-03 |
| CN107224575B CN107224575B (en) | 2018-11-09 |
Family
ID=59932441
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710128121.8A Active CN107224575B (en) | 2017-03-06 | 2017-03-06 | Composition of dairy cow staphylococcus aureus mastitis subunit vaccine and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107224575B (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101182350A (en) * | 2007-11-02 | 2008-05-21 | 山东省农业科学院奶牛研究中心 | Micrococcus pyogenes alpha-hemolysin and coded sequence thereof |
| CN101466406A (en) * | 2006-06-12 | 2009-06-24 | Nabi生物制药公司 | Use of alpha-toxin for treating and preventing staphylococcus infections |
| EP2291196A4 (en) * | 2008-05-12 | 2012-05-30 | Strox Biopharmaceuticals Llc | Staphylococcus aureus-specific antibody preparations |
| CN102698261A (en) * | 2005-06-13 | 2012-10-03 | 葛兰素史密斯克蓝生物品公司 | Use of panton-valentine leukocidin for treating and preventing staphylococcus infections |
| WO2013082558A1 (en) * | 2011-12-02 | 2013-06-06 | Integrated Biotherapeutics, Inc. | Immunogenic composition comprising panton-valentine leukocidin (pvl) derived polypeptides |
| WO2012177658A8 (en) * | 2011-06-19 | 2014-02-20 | New York University | Methods of treating and preventing staphylococcus aureus infections and associated conditions |
| WO2014205111A1 (en) * | 2013-06-19 | 2014-12-24 | Integrated Biotherapeutics, Inc. | Toxoid peptides derived from phenol soluble modulin, delta toxin, superantigens, and fusions thereof |
| WO2015089073A2 (en) * | 2013-12-09 | 2015-06-18 | New York University | Compositions and methods for phagocyte delivery of anti-staphylococcal agents |
| CN105646681A (en) * | 2016-01-21 | 2016-06-08 | 浙江海隆生物科技有限公司 | Preparation method and application of staphylococcus aureus alpha-hemolysin subunit vaccine for dairy cows |
-
2017
- 2017-03-06 CN CN201710128121.8A patent/CN107224575B/en active Active
Patent Citations (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102698261A (en) * | 2005-06-13 | 2012-10-03 | 葛兰素史密斯克蓝生物品公司 | Use of panton-valentine leukocidin for treating and preventing staphylococcus infections |
| CN101466406A (en) * | 2006-06-12 | 2009-06-24 | Nabi生物制药公司 | Use of alpha-toxin for treating and preventing staphylococcus infections |
| CN102743747A (en) * | 2006-06-12 | 2012-10-24 | 葛兰素史密斯克蓝生物品公司 | Use of alpha-toxin for treating and preventing staphylococcus infections |
| CN101182350A (en) * | 2007-11-02 | 2008-05-21 | 山东省农业科学院奶牛研究中心 | Micrococcus pyogenes alpha-hemolysin and coded sequence thereof |
| EP2291196A4 (en) * | 2008-05-12 | 2012-05-30 | Strox Biopharmaceuticals Llc | Staphylococcus aureus-specific antibody preparations |
| WO2012177658A8 (en) * | 2011-06-19 | 2014-02-20 | New York University | Methods of treating and preventing staphylococcus aureus infections and associated conditions |
| CN103717234A (en) * | 2011-06-19 | 2014-04-09 | 纽约大学 | Methods of treating and preventing staphylococcus aureus infections and associated conditions |
| WO2013082558A1 (en) * | 2011-12-02 | 2013-06-06 | Integrated Biotherapeutics, Inc. | Immunogenic composition comprising panton-valentine leukocidin (pvl) derived polypeptides |
| WO2014205111A1 (en) * | 2013-06-19 | 2014-12-24 | Integrated Biotherapeutics, Inc. | Toxoid peptides derived from phenol soluble modulin, delta toxin, superantigens, and fusions thereof |
| WO2015089073A2 (en) * | 2013-12-09 | 2015-06-18 | New York University | Compositions and methods for phagocyte delivery of anti-staphylococcal agents |
| CN105646681A (en) * | 2016-01-21 | 2016-06-08 | 浙江海隆生物科技有限公司 | Preparation method and application of staphylococcus aureus alpha-hemolysin subunit vaccine for dairy cows |
Non-Patent Citations (2)
| Title |
|---|
| HATICE KARAUZUM等: "Structurally Designed Attenuated Subunit Vaccines for S.aureus LukS-PV and LukF-PV Confer Protection in a Mouse Bacteremia Model", 《PLOS ONE》 * |
| 姚蕾: "金黄色葡萄球菌致病毒素的研究进展", 《国际检验医学杂志》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107224575B (en) | 2018-11-09 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Tola et al. | Experimental vaccination against Mycoplasma agalactiae using different inactivated vaccines | |
| CN106754745A (en) | A kind of Friedlander's bacillus bacteriophage and its application | |
| CN110613842B (en) | Subunit vaccine of bovine necrobacillus and preparation method thereof | |
| CN105582524A (en) | Polypeptides and immunizing compositions containing gram positive polypeptides and methods of use | |
| CN103694321B (en) | Streptococcus aureus mSEB mutant and its preparation method and application | |
| CN108126190A (en) | The preparation and application of mycobacteriophage lyases Lysin-Guo1 | |
| Narayanan et al. | Immunogenicity and protective effects of truncated recombinant leukotoxin proteins of Fusobacterium necrophorum in mice | |
| CN102286100B (en) | SEB (staphylococcal enterotoxin B) resisting immune globulin F(ab')2 and preparation method thereof | |
| CN104211804A (en) | Preparation and application of anti-staphylococcus aureus polyclonal antibody | |
| CN103937817B (en) | Newcastle disease virus YT strain, its whole genome sequence and application thereof | |
| CN105861521A (en) | Preparation method and application of tilapia source streptococcus agalactiae recombinant GroEL protein vaccine | |
| CN105646681B (en) | Preparation method and application of staphylococcus aureus alpha-hemolysin subunit vaccine for dairy cows | |
| CN107224575B (en) | Composition of dairy cow staphylococcus aureus mastitis subunit vaccine and preparation method and application thereof | |
| CN107596361A (en) | Subunit vaccine of bovine A-type clostridium perfringens and preparation method and application thereof | |
| CN106310247B (en) | The preparation method of gosling plague inactivated vaccine and its inactivated vaccine of preparation | |
| CN104840424B (en) | A kind of hypericin albumin nanoparticle Escherichia coli serum antibody compound and its preparation method and application | |
| CN116715781A (en) | Recombinant pseudomonas aeruginosa nanoparticle protein reffliC-Ferritin, and preparation method and application thereof | |
| CN106008678A (en) | Fusion protein for inhibiting clostridium perfringens infection and related biological material and application thereof | |
| CN103570835B (en) | Streptococcus aureus ITC fusion rotein and preparation method and application | |
| CN103893760B (en) | Anti- garget drug-fast bacteria IgY and compound bacteriophage composition and preparation method thereof and preparation | |
| CN109106946B (en) | Inactivated staphylococcus aureus vaccine for dairy cow mastitis and preparation method thereof | |
| CN109797139A (en) | 3 type duck hepatitis A virus low virulent strain CH-P60 of one kind and its application | |
| CN105641689A (en) | Preparing method and application of dairy cattle staphylococcus aureus beta-hemolysin subunit vaccine | |
| CN107224576A (en) | Vaccine of staphylococcus aureus mastitis in dairy cows subunit and its preparation method and application | |
| KR101210082B1 (en) | Vaccine composition for swine polyserositis and manufacturing method thereof |
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 | ||
| CP02 | Change in the address of a patent holder |
Address after: 312366 No. 1, Baichuan Road, Binhai New Area, Shaoxing City, Zhejiang Province Patentee after: NOVO BIOTECH Corp. Address before: 312000 5th floor, building 2, science and innovation center, 398 mahuan Road, Binhai New Town, Shaoxing City, Zhejiang Province Patentee before: NOVO BIOTECH Corp. |
|
| CP02 | Change in the address of a patent holder | ||
| CP03 | Change of name, title or address |
Address after: 312366 No. 1, Baichuan Road, Binhai New Area, Shaoxing City, Zhejiang Province Patentee after: Zhejiang Hailong Biotechnology Co.,Ltd. Country or region after: China Address before: 312366 No. 1, Baichuan Road, Binhai New Area, Shaoxing City, Zhejiang Province Patentee before: NOVO BIOTECH Corp. Country or region before: China |
|
| CP03 | Change of name, title or address |