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A kind of anti-foot-and-mouth disease vaccine composition and its preparation method and application

A vaccine composition and foot-and-mouth disease technology, applied to medical preparations containing active ingredients, pharmaceutical formulas, antiviral agents, etc., can solve the problems of easy mutation of foot-and-mouth disease virus, decreased vaccine protection, and weak immunogenicity

Active Publication Date: 2020-11-17
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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. Vaccines designed based on virus-like particles are an ideal form of vaccines. The stability of the structure of FMD virus-like particles is different, which leads to the decrease of activity and cannot induce the body to produce an effective immune response.
[0004] In addition, foot-and-mouth disease virus is prone to mutation, especially the O-type foot-and-mouth disease virus CATHAY variant that is popular in my country. body develops sufficient immunity

Method used

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  • A kind of anti-foot-and-mouth disease vaccine composition and its preparation method and application
  • A kind of anti-foot-and-mouth disease vaccine composition and its preparation method and application
  • A kind of anti-foot-and-mouth disease vaccine composition and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0086] Preparation of Type O Foot-and-Mouth Disease Virus Ⅰ Virus-like Particles (VP4, VP2, VP3, VP1)

[0087] 1. Synthesize the full length of the gene shown in the amino acid sequence of O-type foot-and-mouth disease virus I VP4, VP2, VP3, and VP1 shown in SEQ ID NO.1, 2, 3, and 4 by Sangon Bioengineering (Shanghai) Co., Ltd. The full lengths of the synthesized gene fragments were 255bp, 654bp, 660bp, 639bp, respectively. The foot-and-mouth disease gene template of the present invention is prepared on the basis of the artificially synthesized foot-and-mouth disease gene fragment.

[0088] 2. Construction of foot-and-mouth disease gene expression vector

[0089] For the foot-and-mouth disease gene template synthesized in the previous step, primers were designed respectively (see Table 1), and the O-type foot-and-mouth disease virus I VP4, VP2, VP3, and VP1 genes were amplified.

[0090] Table 1 O-type foot-and-mouth disease virus Ⅰ primer list

[0091]

[0092] Perform ...

Embodiment 2

[0103] Preparation of Type O Foot-and-Mouth Disease Virus Ⅰ Virus-like Particles (VPO, VP3, VP1)

[0104] Referring to the method in Example 1, primers were respectively designed according to the gene sequences of type O foot-and-mouth disease virus I structural proteins VPO, VP3, and VP1 for tandem expression to prepare virus-like particles. The collected bacteria were resuspended according to the ratio of 1 g of bacteria to 10 ml of lysate, and the bacteria were crushed 4 times with a homogenizer at a pressure of 800 bar. Centrifuge at 13500rpm for 40min, save the supernatant, and detect by 15% SDS-PAGE electrophoresis. At this time, the expression level of the three serially expressed proteins in the supernatant is about 20%. Ammonium sulfate fractional precipitation was used for crude protein purification, followed by chromatographic purification. The purified protein was subjected to SDS-PAGE electrophoresis, which showed that the target protein was purified and enriched....

Embodiment 3

[0107] Preparation of Asian Foot-and-Mouth Disease Type 1 Virus-like Particles (VP0, VP3, VP1)

[0108] The genes shown in the amino acid sequences of Asian type 1 foot-and-mouth disease virus VPO, VP3, and VP1 shown in SEQ ID NO.21, 22, and 23 were respectively designed according to the method of Example 1 and expressed in tandem to prepare virus-like particles.

[0109] Negative staining with phosphotungstic acid and electron microscope observation showed that the FMD protein had formed virus-like particles, and the formed virus-like particles were plump, with high assembly efficiency and no aggregation. After placing the foot-and-mouth disease virus-like particles at 4°C for 4 months, negative staining with phosphotungstic acid and electron microscope observation showed that the virus-like particles were still full without aggregation. It shows that the foot-and-mouth disease protein prepared according to the three-stage expression of the screened sequence of the present in...

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Abstract

The invention discloses a foot-and-mouth disease virus-like particle, a preparation method, a vaccine composition and application. The vaccine composition of the present invention is composed of O-type foot-and-mouth disease virus-like particles composed of foot-and-mouth disease virus structural proteins VP4, VP2, VP3, and VP1 and / or Asian type 1 foot-and-mouth disease virus-like particles and / or foot-and-mouth disease virus-like particles composed of foot-and-mouth disease structural proteins VPO, VP3, and VP1 The type A foot-and-mouth disease virus-like particles composed of structural proteins VP0, VP3, and VP1 are composed of an adjuvant, which has a stable structure and reasonable components. It can quickly form specific antibodies, significantly increase the duration of immunity, and maintain long-term immune protection.

Description

technical field [0001] The invention relates to the field of veterinary biological products, more specifically, the invention relates to foot-and-mouth disease virus-like particles, a preparation method, and a vaccine composition prepared from the virus-like particles. Background technique [0002] Foot-and-mouth disease (FMD) is an acute, highly contagious animal disease that can spread quickly and over long distances. It is the most contagious disease among mammals, and the infection of artiodactyls will cause significant economic losses worldwide. Animals affected by FMD include cattle, sheep, goats and pigs. The causative agent, foot-and-mouth disease virus (FMDV), is an aphthous sore virus of the picornavirus family. The virus is divided into 7 serotypes (A, O, C, Asia1, SAT1, SAT2, and SAT3), among which type O, Asia1 and A are mainly prevalent in my country. Vaccine immunization is an effective measure to control the disease and protect livestock from harm. [0003...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K39/135A61K39/39A61P31/14
Inventor 张许科袁于人孙进忠陈红英肖燕田克恭
Owner PU LIKE BIO ENG
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