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1. (WO2019023920) FOOT-AND-MOUTH DISEASE VIRUS-LIKE PARTICLE VACCINE AND PREPARATION METHOD THEREFOR
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Pub. No.: WO/2019/023920 International Application No.: PCT/CN2017/095376
Publication Date: 07.02.2019 International Filing Date: 01.08.2017
IPC:
C12N 15/42 (2006.01) ,C12N 15/63 (2006.01) ,A61K 39/135 (2006.01) ,A61P 31/14 (2006.01)
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
11
DNA or RNA fragments; Modified forms thereof
31
Genes encoding microbial proteins, e.g. enterotoxins
33
Genes encoding viral proteins
40
Proteins from RNA viruses, e.g. flaviviruses
41
Picornaviridae, e.g. rhinovirus, coxsackie viruses, echoviruses, enteroviruses
42
Foot-and-mouth disease virus
C CHEMISTRY; METALLURGY
12
BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
N
MICRO-ORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICRO-ORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
15
Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
09
Recombinant DNA-technology
63
Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
A HUMAN NECESSITIES
61
MEDICAL OR VETERINARY SCIENCE; HYGIENE
K
PREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
39
Medicinal preparations containing antigens or antibodies
12
Viral antigens
125
Picornaviridae, e.g. calicivirus
135
Foot-and-mouth disease virus
A HUMAN NECESSITIES
61
MEDICAL OR VETERINARY SCIENCE; HYGIENE
P
SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
31
Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
12
Antivirals
14
for RNA viruses
Applicants:
金普诺安生物科技(苏州)有限公司 GPROAN BIOTECHNOLOGIES (SUZHOU), INC. [CN/CN]; 中国江苏省太仓市 生物医药产业园灵溪路18号 No.18 Lingxi Rd., Taicang Biomedical Park Taicang, Jiangsu 215425, CN
Inventors:
安海谦 AN, Haiqian; CN
冉波 RAN, Bo; CN
赵荣茂 ZHAO, Rongmao; CN
方华明 FANG, Huaming; CN
任玉珍 REN, Yuzhen; CN
王鹏 WANG, Peng; CN
Agent:
北京路浩知识产权代理有限公司 CN-KNOWHOW INTELLECTUAL PROPERTY AGENT LIMITED; 中国北京市 海淀区丹棱街3号中国电子大厦B座18层 18th Floor, Tower B, CEC Plaza No.3 Dan Ling Street, Haidian District Beijing 100080, CN
Priority Data:
Title (EN) FOOT-AND-MOUTH DISEASE VIRUS-LIKE PARTICLE VACCINE AND PREPARATION METHOD THEREFOR
(FR) VACCIN À PARTICULES DE TYPE VIRUS DE LA FIÈVRE APHTEUSE ET SON PROCÉDÉ DE PRÉPARATION
(ZH) 一种口蹄疫病毒样颗粒疫苗及其制备方法
Abstract:
(EN) The present invention provides a foot-and-mouth disease virus-like particle vaccine and a preparation method therefor. The preparation method comprises: constructing a recombinant expression plasmid by inserting two ORFs of P12A and 3C genes of a codon-optimized FMDV in series into a transfer vector; or constructing a recombinant expression plasmid by cloning a single ORF containing the P12A3C gene into the vector under promoter SV40, transforming a competent cell to obtain recombinant Bacmid-DNA, transfecting sf9 cells, so as to obtain an insect cell capable of expressing foot-and-mouth disease virus-like granule proteins. After the cells have been cultured, the supernatant is collected, and the VLP is purified by means of ultrafiltration, centrifugation and chromatography to prepare a foot-and-mouth disease virus-like particle vaccine.
(FR) La présente invention concerne un vaccin à particules de type virus de la fièvre aphteuse et son procédé de préparation. Le procédé de préparation comprend : la construction d’un plasmide d’expression recombinant par insertion de deux ORF des gènes P12A et 3C d’un FMDV à codon optimisé en série dans un vecteur de transfert ; ou la construction d’un plasmide d’expression recombinant par clonage d’un ORF unique contenant le gène P12A3C dans le vecteur sous le promoteur du SV40, la transformation d’une cellule compétente pour obtenir l’ADN-Bacmid recombinant, la transfection des cellules sf9, afin d’obtenir une cellule d’insecte capable d’exprimer les protéines en granule de type virus de la fièvre aphteuse. Après que les cellules ont été cultivées, le surnageant est recueilli, et la VLP est purifiée par ultrafiltration, par centrifugation et par chromatographie pour préparer un vaccin à particules de type virus de la fièvre aphteuse.
(ZH) 本发明提供了一种口蹄疫病毒样颗粒疫苗及其制备方法。将密码子优化的FMDV的P12A和3C基因两个ORF串联插入转移载体构建重组表达质粒;或通过启动子SV40下的包含P12A3C基因的单一ORF克隆入载体构建重组表达质粒,转化感受态细胞得到重组Bacmid-DNA,转染sf9细胞,得到能够表达口蹄疫病毒样颗粒蛋白的昆虫细胞。将该细胞培养后收集上清,超滤浓缩、离心、层析纯化VLP,制备口蹄疫病毒样颗粒疫苗。
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Designated States: AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW
African Regional Intellectual Property Organization (ARIPO) (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW)
Eurasian Patent Office (AM, AZ, BY, KG, KZ, RU, TJ, TM)
European Patent Office (EPO) (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR)
African Intellectual Property Organization (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG)
Publication Language: Chinese (ZH)
Filing Language: Chinese (ZH)