Preparation method of phage display-expressing circovirus antigen vaccine

A phage display, circovirus technology, applied in the direction of viral antigen components, antiviral immunoglobulins, viral peptides, etc., can solve the problems of complex methods, little effect and high cost, and achieve easy preparation, good protection effect, The effect of low production cost
CN104474540AInactive Publication Date: 2015-04-01SHANDONG SINDER TECH
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
SHANDONG SINDER TECH
Publication Date
2015-04-01
Estimated Expiration
Not applicable · inactive patent

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Abstract

The invention provides a preparation method of a phage display-expressing circovirus antigen vaccine. The preparation method comprises the following steps: extracting porcine circovirus II type (PCV2) RNA, carrying out inverse transcription, carrying out a polymerase chain reaction (PCR) for amplification so as to obtain porcine II type circovirus ORF2 gene's open reading frame fragments, directionally inserting the fragments into a T4 phage; obtaining a T4 phage with the ORF2 gene by an immunoscreening method; purifying and infecting X-Blue ST1(CCTCC M 2014397) host cell again; carrying out fermental cultivation to obtain the X-Blue ST1 host cell, and carrying out inducible expression and inactivation and taking a culture as an antigen; carrying out ELISA calibration of antigen valence, and carrying out double dilution on the antigen with normal saline to the final concentration of 107U / ml; and a vaccine adjuvant is added according to the ratio of 1:2.5, so as to obtain the vaccine. According to the invention, the phage is utilized to express the porcine circovirus II type antigen and the vaccine is prepared. Thus, the virulence enhancement risk of an attenuated vaccine is avoided. In addition, fermental cultivation is beneficial to large-scale industrial production. Cost of the vaccine provided by the invention is far lower than that of a vaccine produced by a cell culture method.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and in particular relates to a preparation method of a circovirus antigen vaccine expressed by phage display. Background technique

[0002] Porcine circovirus infection is an acute infectious disease of pigs caused by circovirus. The main features are emaciation, decreased physical fitness, anemia, jaundice, diarrhea, stunted growth, dyspnea, and reproductive impairment in sows. with extensive pathology. According to the different serotypes, it can be divided into PCV1 and PCV2. Among them, PCV2 is the most serious infection reported in my country. Currently, the methods for treating PCV2 are complex and costly, and the technical effects and gains are minimal. Contents of the invention

[0003] The purpose of the present invention is to aim at the deficiencies of the prior art, and provide a preparation method of a circovirus antigen vaccine expressed by phage display, so as to control the infe...

Claims

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