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Porcine foot-and-mouth disease virus type a fc polypeptide vaccine and its preparation method and application

A porcine foot-and-mouth disease virus and polypeptide vaccine technology, applied in porcine foot-and-mouth disease virus type A Fc polypeptide vaccine and its preparation, porcine foot-and-mouth disease vaccine and its preparation field, can solve the problems of product lag and long development cycle, and achieve strong immune efficacy and high protection Sexual antibodies, the effect of small differences in antibody levels in immunized animals

Active Publication Date: 2022-03-25
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

In order to quickly develop new vaccines for the prevention and control of newly introduced foot-and-mouth disease viruses, overcome the long development cycle and product lag of traditional inactivated vaccines, further improve the immune effect of epitope vaccines, and reduce the gap with inactivated vaccines

Method used

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  • Porcine foot-and-mouth disease virus type a fc polypeptide vaccine and its preparation method and application
  • Porcine foot-and-mouth disease virus type a fc polypeptide vaccine and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1. Design and synthesis of swine foot-and-mouth disease virus type A Fc polypeptide

[0025] 1. Design of swine foot-and-mouth disease virus type A polyepitope

[0026] According to the main antigen genes of the three topological strains of swine foot-and-mouth disease virus type A (AF / 72, A / HB / WH / 09, A / GDMM / 2013), the amino acid sequences of 140-160 and 200-213 in the coding region of VP1 gene were selected The amino acid sequence was used as an antigenic epitope, which was then concatenated in the appropriate order, i.e. 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 concatenating epitopes, spacer sequences are introduced between adjacent epitopes to ensure the independence of epitopes. The spacers are GGSSGG, GPLS and GGGS. The nucleotide sequence of the obtained swine foot-and-mouth disease virus type A polyepitope is shown in SEQ ID NO.1.

[0027] 2. ...

Embodiment 2

[0031] Example 2. Preparation of Porcine Foot-and-Mouth Disease Virus Type A Fc Polypeptide Vaccine

[0032] The purified swine foot-and-mouth disease virus type A Fc polypeptide prepared in Example 1 was quantified by Bio-Rad quantitative kit and diluted to an appropriate concentration, and the oil adjuvant Montanide ISA206 (Montanide ISA206 ( Seppic, France) was emulsified into a vaccine preparation (W / O / W), 1 ml per head, containing 200 μg of soluble swine foot-and-mouth disease virus type A Fc polypeptide.

Embodiment 3

[0033] Example 3. Immune efficacy test

[0034] The pigs used in the experiment weighed about 40kg, the antibody of type A foot-and-mouth disease virus was less than 1:4 (liquid phase blocking ELISA result), and the 3ABC protein antibody was negative (3ABC antibody chemiluminescence kit result). Five pigs (containing 200 μg of soluble antigen) were inoculated with the Fc polypeptide vaccine of the present invention (prepared in Example 2) by intramuscular route of 1 ml per head. 21 days after priming, all pigs were boosted once with the same dose of vaccine. 14 days after booster immunization, the antibody titer of each pig was determined, together with 3 control pigs with equal conditions in the ABSL-3 laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, in accordance with the national standard with type A foot-and-mouth disease virus (A / GDMM / 2013 strains) were challenged to pigs and observed for 10 consecutive days. The results sho...

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Abstract

The invention discloses a swine foot-and-mouth disease virus type A Fc polypeptide vaccine and a preparation method and application thereof. The present invention takes the Fc of swine IgG as the skeleton, combined with the current situation of swine A-type foot-and-mouth disease epidemic and the prevention and control requirements in my country, and uses reverse vaccinology, bioinformatics and biochemical knots and other technologies to screen and design different topological types of swine foot-and-mouth disease A. The epitope antigen gene of the virus strain is used to display the antigenic epitope in the functional region of the immunoglobulin Fc gene, and a swine foot-and-mouth disease type A Fc polypeptide vaccine suitable for the epidemic situation and prevention and control needs of my country has been developed. And by screening the prokaryotic expression system and optimizing the expression conditions, the soluble expression of the fusion protein was realized, and the antigenic epitope was displayed to the greatest extent. Experiments have proved that the swine foot-and-mouth disease type A Fc polypeptide vaccine of the present invention has stronger immune efficacy, and can induce the body to produce higher protective antibodies after boosting immunization; the difference in antibody levels of immunized animals is small, and the immunized pigs are highly virulent. After attack, 100% get protection.

Description

technical field [0001] The invention relates to a swine foot-and-mouth disease vaccine and a preparation method and application thereof, in particular to a swine foot-and-mouth disease virus type A Fc polypeptide vaccine, a preparation method and application thereof, and 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 exports. Once the epidemic occurs, the losses will be heavy and the impact will be severe. The World Organization for Animal Health (OIE) lists it as a notifiable infectious disease, and my country lists it as a Class I 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 Plan (2012-2020)" and "National Foot-and...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00C12N15/42C12N15/62A61K39/135A61P31/14
CPCA61K39/12C07K14/005C07K2319/30C12N2770/32134C12N2770/32122
Inventor 常惠芸李杨帆邵军军张永光
Owner LANZHOU INST OF VETERINARY SCI CHINESE ACAD OF AGRI SCI
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