Pig foot-and-mouth disease virus O type and A type Fc polypeptide bivalent vaccine, as well as preparation method and application thereof

A porcine foot-and-mouth disease virus and vaccine technology, applied in the field of medicine, can solve problems such as unsatisfactory requirements

Active Publication Date: 2018-07-13
LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the development of inactivated foot-and-mouth disease vaccines by classical methods can no

Method used

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  • Pig foot-and-mouth disease virus O type and A type Fc polypeptide bivalent vaccine, as well as preparation method and application thereof
  • Pig foot-and-mouth disease virus O type and A type Fc polypeptide bivalent vaccine, as well as preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] Embodiment 1, design and synthesis of porcine foot-and-mouth disease virus type A Fc polypeptide

[0029] 1. Design of porcine foot-and-mouth disease virus type A multi-epitope gene

[0030]According to the main antigen genes of three topological strains of porcine foot-and-mouth disease virus A (AF / 72, A / HB / WH / 09, A / GDMM / 2013), the 140-160 amino acid sequence and 200-213 amino acid sequence of the VP1 gene coding region were selected. The amino acid sequence is used as the epitope, and then the appropriate order is selected for tandem, that is, 140-160(A / GDMM / 2013)-GGSSGG-140-160(A / HBWH / 09)-GPLS-140-160(AF / 72)-GGGS-200-213 (A / GDMM / 2013), in order to prevent the formation of new epitopes after tandem epitopes, a spacer sequence is introduced between adjacent epitopes to ensure the independence of the epitopes. The spacers are GGSSGG, GPLS, and GGGS. The nucleotide sequence of the obtained porcine foot-and-mouth disease virus type A multi-epitope gene is shown in SEQ ...

Embodiment 2

[0035] Cloning and protein expression thereof of the gene of porcine foot-and-mouth disease virus O-type Fc polypeptide of embodiment 2, purification

[0036] 1. Optimization of porcine foot-and-mouth disease virus type O broad-spectrum multi-epitope fusion gene

[0037] Using bioinformatics code optimization software combined with multidisciplinary technologies such as immunology and biochemistry, the patent application with the publication number CN102675471A and the invention title "Porcine Foot-and-Mouth Disease Virus Type O Broad-spectrum Multi-epitope Recombinant Antigen and Its Application" was disclosed The nucleotide sequence encoding porcine foot-and-mouth disease virus type O broad-spectrum multi-epitope recombinant antigen was codon-optimized, and the optimized nucleotide sequence was shown as SEQ ID NO.4, named MEO-Fc. The sequence comparison analysis (Clustal W software and DNAMEN Version 9) of the genes before and after optimization showed that there were large ...

Embodiment 3

[0041] Embodiment 3, the preparation of porcine foot-and-mouth disease virus O type, A type Fc polypeptide bivalent vaccine

[0042] 1. Vaccine preparation

[0043] The two recombinant proteins MEA-Fc and MEO-Fc purified in Examples 1 and 2 were quantified by the Bio-Rad quantitative kit and then diluted to an appropriate concentration. The two proteins were prepared according to MEA-Fc:MEO-Fc=2: 1 (w / w) mixed, emulsified with oil adjuvant Montanide ISA206 (Seppic, France) according to 1:1 (w / w) into a vaccine preparation (W / O / W), 1ml per head, which contains porcine foot-and-mouth disease Viral O-type Fc polypeptide MEO-Fc 100 μg, porcine foot-and-mouth disease virus A-type Fc polypeptide MEA-Fc 200 μg.

[0044] 2. Immunity efficacy test:

[0045] The pigs used in the experiment weighed about 40kg, the O-type and A-type foot-and-mouth disease virus antibodies were less than 1:4 (results of liquid-phase blocking ELISA), and the 3ABC protein antibodies were negative (results ...

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Abstract

The invention discloses a pig foot-and-mouth disease virus O type and A type Fc polypeptide bivalent vaccine, as well as a preparation method and application thereof. The bivalent vaccine is preparedfrom effective components of pig foot-and-mouth disease virus A type Fc polypeptide and pig foot-and-mouth disease virus O type Fc polypeptide, wherein an amino acid sequence of the pig foot-and-mouthdisease virus A type Fc polypeptide is as shown in SEQ ID NO.3; an amino acid sequence of the pig foot-and-mouth disease virus O type Fc polypeptide is as shown in SEQ ID NO.5. Experimental results show that the bivalent vaccine can induce a body to produce a high-level neutralizing antibody against O type and A type foot-and-mouth disease viruses after being subjected to booster immunization, can protect immunized pigs against the attack of the O type and A type foot-and-mouth disease viruses, and realizes 5/5 protection (i.e. the protection rate is 100 percent). The pig foot-and-mouth disease virus O type and A type Fc polypeptide bivalent vaccine, as well as the preparation method and the application thereof provided by the invention provide a technical support and vaccine storage forguaranteeing healthy sustainable development of animal husbandry in China and realizing a mid-and-long-term prevention and control aim of foot-and-mouth disease countries.

Description

technical field [0001] The invention relates to a porcine foot-and-mouth disease vaccine and its preparation method and application, in particular to a porcine foot-and-mouth disease virus O-type and A-type Fc polypeptide bivalent vaccine and its preparation method and application. The invention belongs to the technical field of medicine. Background technique [0002] As a major animal disease, foot-and-mouth disease not only seriously threatens the healthy development of animal husbandry, but also involves the safety of animal-derived food and its foreign trade export. Once an epidemic occurs, the losses will be heavy and the impact will be severe. The World Organization for Animal Health (OIE) lists it as an infectious disease that must be notified, and my country lists it as a first-class animal infectious disease. In order to purify and eliminate foot-and-mouth disease, my country has formulated the "National Medium- and Long-Term Animal Disease Prevention and Control P...

Claims

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Application Information

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IPC IPC(8): A61K39/135A61K39/295A61P31/14
CPCA61K39/12A61K2039/552A61K2039/575A61K2039/70C12N2770/32134
Inventor 常惠芸邵军军李杨帆张永光
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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