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Subunit vaccine for infectious bursal disease of chicken and preparation method thereof

A subunit vaccine and chicken infectious technology, applied in the field of chicken infectious bursal disease subunit vaccine, can solve the problems of low immune efficacy, difficult to solve the persistent infection of bursal disease degree, immune failure, etc., to promote synergy Synergistic effects, overcoming biological activity problems, enhancing immunogenicity

Inactive Publication Date: 2018-11-16
TIANJIN RINGPU BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical purpose of the present invention is to provide a chicken-infected bursa VP2 subunit vaccine with high immune efficacy, so as to solve the problem in the prior art that the immune efficacy of the VP2 subunit vaccine is low and difficult to be used clinically and popularized
[0006] Another technical purpose of the present invention is to overcome problems such as the blank period of immunity in traditional vaccines, the difficulty of one-time immunization to solve the persistent infection of bursal disease, and the failure of immunity caused by virus mutation during the course of the disease.

Method used

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  • Subunit vaccine for infectious bursal disease of chicken and preparation method thereof
  • Subunit vaccine for infectious bursal disease of chicken and preparation method thereof
  • Subunit vaccine for infectious bursal disease of chicken and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1 Recombinant protein expression

[0027] 1) VP2 prokaryotic protein expression

[0028] Utilizing the chicken bursal virus VP2 gene in GenBank, a specific primer was designed to amplify the target fragment; this fragment was cloned into the pMD18-T vector to obtain the recombinant cloning vector pMD18-T-VP2; , The VP2 gene on the recombinant cloning vector with the correct sequence after sequencing identification is subcloned to the prokaryotic expression vector pET-28a to obtain the recombinant expression vector pET-28a-VP2; the recombinant expression vector is transformed into Escherichia coli expression host (BL21), The expressed product is purified and detected to obtain the VP2 protein.

[0029] 2) ChIFNα, ChIL2 eukaryotic protein expression

[0030] Refer to the expression method of "VP2 prokaryotic protein expression" to obtain the pFastBacDual-IFNα and pFastBacDual-ChIL recombinant expression vectors respectively; after obtaining the recombinant expre...

Embodiment 2

[0035] Example 2 Preparation of Genetic Engineering Subunit Vaccine

[0036] 1) Preparation of VP2 protein subunit vaccine

[0037] The VP2 protein was diluted with physiological saline to a protein content of 0.5 mg / ml, and then the aluminum gel adjuvant and the diluted protein were mixed uniformly at a ratio of 1:4 (volume ratio) to prepare a VP2 subunit vaccine for use.

[0038] 2) Preparation of ChIFNα-VP2 protein subunit vaccine

[0039] Dilute the VP2 protein with normal saline to a protein content of 0.5 mg / ml, add ChIFNα protein with a final content of 500 ng / ml, stir and mix evenly, and mix the aluminum gel adjuvant and the above mixed protein at a ratio of 1:4 (V / V) Prepare chicken ChIFNα-VP2 subunit vaccine for use.

[0040] 3) Preparation of ChIL2-VP2 protein subunit vaccine

[0041] Dilute the VP2 protein with normal saline to a protein content of 0.5 mg / ml, add ChIL2 protein with a final content of 500 ng / ml, stir and mix evenly, then mix the aluminum gel adjuva...

Embodiment 3

[0048] Example 3 Detection of serum antibody levels after immunization

[0049] Take 14-day-old SPF chickens and divide them into 7 groups randomly, with 10 chickens in the first group and 20 chickens in the remaining groups, and carry out the following immunization procedures respectively.

[0050] The first group: as a blank control group, each intramuscular injection of 0.15ml of normal saline;

[0051] The second group: VP2 protein subunit vaccine group, each intramuscular injection of VP2 protein subunit vaccine 0.15ml;

[0052] The third group: ChIFNα-VP2 protein subunit vaccine group, each intramuscular injection of ChIFNα-VP2 protein subunit vaccine 0.15ml;

[0053] The fourth group: ChIL2-VP2 protein subunit vaccine group, each intramuscular injection of ChIL2-VP2 protein subunit vaccine 0.15ml;

[0054] The fifth group: ChIFNα-ChIL2-VP2 protein subunit vaccine group, each intramuscular injection of ChIFNα-ChIL2-VP2 protein subunit vaccine 0.15ml;

[0055] The sixt...

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Abstract

The invention provides genetic engineering subunit vaccine of infectious bursal disease of chicken which is composed of the eukaryotic co-expression product of the genes of IFN-alpha and IL-2 of chicken, the prokaryotic expression product of the virus VP2 of the infectious bursal disease of chicken and aluminum oxide adjuvant, and aims at solving the problems of low immune efficacy, difficult of clinical use and popularization of the VP2 genetic engineering subunit vaccine in the prior art.

Description

technical field [0001] The present invention relates to the field of genetic engineering subunit vaccines, in particular to a chicken infectious bursal disease subunit vaccine, in particular to a co-expression product of chicken interleukin-2 and chicken alpha interferon to increase chicken bursa VP2 Subunit vaccines. Background technique [0002] Genetic engineering subunit vaccine is the crystallization of modern genetic engineering technology. With its unparalleled high safety, stability and low production cost, it has become the favorite in the field of modern vaccine preparation. However, due to the poor immunogenicity of its antigen A congenital deficiency makes it impossible to completely ban traditional vaccines. Therefore, how to improve the immunogenicity of genetically engineered subunit vaccine antigens or enhance their immune efficacy has become the focus of research in recent years. The use of cytokines with immune regulation and immune effector functions as ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/39A61K39/12A61P31/14
CPCA61K39/12A61K39/39A61K2039/552A61K2039/55505A61K2039/55516A61P31/14C12N2720/10034
Inventor 李守军孙晨王甜田凤菊郁宏伟康亚男陈冰李亚杰
Owner TIANJIN RINGPU BIO TECH
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