O-type foot-and-mouth disease virus-like particle antigen, vaccine composition containing O-type foot-and-mouth disease virus-like particle antigen, and preparation method and application of vaccine composition

A technology of foot-and-mouth disease virus and vaccine composition, which is applied in the field of O-type foot-and-mouth disease virus-like particle antigens, can solve the problems of no formation, poor vaccine immune effect, and inability to effectively exert the immunogenicity of VLPs vaccines, and achieve good stability and maintenance The effect of antibody titer and immune effect

Active Publication Date: 2020-06-05
PU LIKE BIO ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

VLPs vaccines can effectively stimulate the body to produce anti-infection and anti-tumor immunity, but VLPs with a stable structure cannot be formed, that is, the immunogenicity of VLPs vaccines cannot be effectively exerted, resulting in poor vaccine immunity
There are no successful examples of VLPs vaccine against FMD Ind strain O

Method used

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  • O-type foot-and-mouth disease virus-like particle antigen, vaccine composition containing O-type foot-and-mouth disease virus-like particle antigen, and preparation method and application of vaccine composition
  • O-type foot-and-mouth disease virus-like particle antigen, vaccine composition containing O-type foot-and-mouth disease virus-like particle antigen, and preparation method and application of vaccine composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 Expression of O-type Ind strain foot-and-mouth disease virus-like particles

[0046] 1. Construction of expression vector

[0047] The VPO gene fragment shown in the sequence table SEQ ID NO.1, the VP3 gene fragment shown in the sequence table SEQ ID NO.2, and the VP1 gene fragment shown in the sequence table SEQ ID NO.3 were synthesized by Jinweizhi Company, respectively, and pBLUE -T Vector vector connection, connect the successfully connected recombinant clones to the pET28a vector after the same restriction digestion, The positive clone pET28a-VP0-VP3-VP1 inserted into O-type Ind strain foot-and-mouth disease virus VPO, VP3, VP1 genes was obtained. Transform the ligated plasmid into CaCl 2 The prepared DH5α competent cells were spread on the kanamycin-resistant solid LB medium, and when the monoclonal colony was clearly visible, the single clone was picked into the LB liquid medium containing kanamycin, 37 C, 230 rev / min, cultivate overnight for 12 hour...

Embodiment 2

[0055] The preparation of embodiment 2 O type Ind strain foot-and-mouth disease virus-like particle vaccine composition

[0056] Take the virus-like particles prepared in Example 1 and slowly add them to the adjuvant. During the process of adding, stir continuously with an emulsifier with a rotation speed of 800 rpm for 12 minutes, mix well, and store at 4° C., which is the foot-and-mouth disease virus-like particle vaccine composition. See Table 1 for specific ratios. The adjuvants suitable for the present invention may be adjuvants known to those skilled in the art. In this example, the adjuvant ISA 206 (Seppic, France) was selected.

[0057] Table 1 O-type Ind strain foot-and-mouth disease virus-like particle vaccine composition ratio

[0058] components Vaccine 1 Vaccine 2 Vaccine 3 FMD antigen (μg / ml) 160 200 240 ISA 206 Adjuvant (V / V%) 50％ 50％ 50％

Embodiment 3

[0059] Example 3 The immunogenicity test of the O-type Ind strain foot-and-mouth disease virus-like particle vaccine composition

[0060] Select 20 healthy susceptible piglets with a weight of about 40kg, negative for type O foot-and-mouth disease virus antigen and antibody, and divide them into 4 groups randomly, 5 pigs in each group. Groups 1-3 are vaccine 1, vaccine 2, and vaccine 3 immunization groups prepared in Example 2 of the present invention, respectively, and group 4 is a control group. The way of immunization in the immunized group was intramuscular injection of 2 ml in the neck, and the control group was immunized with the same amount of PBS. Blood was collected from each pig before vaccine immunization, and blood was collected on the 7th, 14th, 21st, and 28th days after immunization.

[0061] The collected sera were tested for related antibodies using the Type O Ind strain foot-and-mouth disease antibody ELISA detection kit. The results showed that the antibodi...

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Abstract

The invention provides a foot-and-mouth disease virus-like particle antigen. The foot-and-mouth disease virus-like particle antigen consists of a VP0 antigen protein, a VP1 antigen protein and a VP3 antigen protein through assembling, wherein the VP0 antigen protein is coded by a nucleotide sequence as shown in Seq ID No.1 or coded by a degenerate sequence of the nucleotide sequence as shown in Seq ID No.1, the VP1 antigen protein is coded by a nucleotide sequence as shown in Seq ID No.3 or coded by a degenerate sequence of the nucleotide sequence as shown in Seq ID No.3, and the VP3 antigen protein is coded by a nucleotide sequence as shown in Seq ID No.2 or coded by a degenerate sequence of the nucleotide sequence as shown in Seq ID No.2. The foot-and-mouth disease virus-like particle antigen has favorable immunogenicity, a vaccine composition prepared from the foot-and-mouth disease virus-like particle antigen can protect current epidemic O-type Ind strains through one-time immunization, and the immunoprotection period is long. The invention further provides the vaccine composition prepared from the foot-and-mouth disease virus-like particle antigen, and a preparation method andan application of the vaccine.

Description

technical field [0001] The present invention belongs to viral antigen and the pharmaceutical preparation of the antigen. Specifically, the present invention relates to an O-type foot-and-mouth disease virus-like particle antigen, a vaccine composition prepared therefrom, a preparation method and application thereof. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, highly contagious animal disease that can spread quickly and over long distances. Significant economic losses worldwide. Animals affected by FMD include cattle, sheep and pigs. The causative agent is foot-and-mouth disease virus (FMDV), an aphthous sore virus of the picornavirus family. The virus is divided into seven serotypes (A, O, C, Asial, SAT1, SAT2 and SAT3), among which the O-type FMD virus is the most prevalent. Vaccine immunization is an effective measure to control the disease and protect livestock from harm. However, there is no vaccine against the O-type new Ind strain circu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/09C12N15/42C12N15/70A61K39/135A61P31/14
CPCC07K14/005C12N15/70A61K39/12A61P31/14C12N2770/32122C12N2770/32134A61K2039/5258A61K2039/552A61K2039/54Y02A50/30
Inventor 田克恭张素玲孙进忠张许科
Owner PU LIKE BIO ENG
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