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Classical swine fever E2 subunit vaccine and application thereof

A subunit vaccine, swine fever technology, applied in applications, medical preparations containing active ingredients, viral peptides, etc., can solve the problems of reduced safety and effectiveness, and achieve stable effective antigen quantity, good safety, antigenic high purity effect

Inactive Publication Date: 2016-11-23
北京大北农科技集团股份有限公司动物医学研究中心 +3
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The vaccine prepared by it uses the expressed supernatant to directly immunize animals. The protein has not been purified, and there will be many foreign proteins and culture media in it, which greatly reduces the safety and effectiveness

Method used

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  • Classical swine fever E2 subunit vaccine and application thereof
  • Classical swine fever E2 subunit vaccine and application thereof
  • Classical swine fever E2 subunit vaccine and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Embodiment 1 preparation of classical swine fever virus E2 protein

[0030] 1) Optimization and synthesis of classical swine fever virus E2 gene

[0031] Referring to the E2 gene of classic strains and popular strains at home and abroad, select the classical swine fever virus strain of genotype 2.1 (gene sequence is Classical swine fever virus strain Zj0801, complete genome GenBank: FJ529205.1) according to the codon preference in insect cells The E2 gene of classical swine fever virus was optimized and handed over to the company for synthesis. The artificially synthesized genes include: GP67 signal peptide nucleic acid sequence (129bp), partial sequence of classical swine fever virus E2 gene (1011bp), 6His tag sequence (18bp) and fragment 5 EcoR I restriction site (6bp) at the 'end and Xho I restriction site (6bp) at the 3' end of the fragment (see figure 1 ) (shown in SEQ ID No.2).

[0032] 2) Cloning of the gene:

[0033] The artificially synthesized exogenous DNA...

Embodiment 2

[0041] Example 2 Preparation of classical swine fever virus E2 protein subunit vaccine

[0042] E2 protein of classical swine fever virus is emulsified with 201R to make vaccine: preparation of adjuvant, 201R adjuvant (121°C, 15pa, 20min) is sterilized, protein antigen and adjuvant are mixed 1:1, emulsified at 350rpm for 5min, namely The recombinant subunit vaccine of classical swine fever E2 was prepared.

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Abstract

The invention provides a classical swine fever E2 subunit vaccine and a preparation method thereof. The preparation method specifically comprises the steps that a Baculovirus Carrier Expression System is used for expressing a large amount of recombination classical swing fever E2 protein in insect cells, and the classical swine fever E2 subunit vaccine with a good immune effect is developed. The Bac-to-Bac Baculovirus Carrier Expression System is used for expressing the classical swine fever E2 protein. The classical swine fever E2 protein and an adjuvant 201R are emulsified into the novel subunit vaccine, and by means of piglet immune challenge tests, it is verified that the subunit vaccine has a good immune protection effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a swine fever E2 subunit vaccine and a preparation method thereof. Background technique [0002] Classical Swine Fever (CSF) is a highly contagious disease caused by Classical Swine Fever Virus (CSFV), and pigs are the only natural host of the virus. Its popularity has spread all over the world for nearly a hundred years. The International Office of Epizootics defines it as a Class A infectious disease, and it is one of the main diseases that endanger the development of my country's pig industry. Vaccination is the main measure for the prevention and control of swine fever in developing countries. At present, the most widely used attenuated classical swine fever vaccines include Chinese classical swine fever rabinized attenuated vaccine (strain C), Japanese GPE-vaccine and French Thiverval vaccine. However, in recent years, the quality of swine fever vaccine is difficult to guarante...

Claims

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Application Information

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IPC IPC(8): A61K39/12A61K39/39C12N15/40C12N15/866C07K14/18A61P31/14
Inventor 石艳丽张国庆刘长辉周景云刘培培王贵华赵亚荣
Owner 北京大北农科技集团股份有限公司动物医学研究中心
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