Preparation method of novel genetically recombinant pure protein antigen

A protein antigen, gene recombination technology, applied in the direction of recombinant DNA technology, antiviral immunoglobulin, medical preparations containing active ingredients, etc. high purity effect

Inactive Publication Date: 2014-07-02
SHANGHAI SIQI MEDICAL TECH
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Problems solved by technology

H7N9 avian influenza has caused a large number of infection cases in East China and other regions. At present, there is no effective prevention and treatment method. The H7N9 vaccines under research include inactivated and attenuated H7N9 viruses, ge

Method used

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Embodiment Construction

[0006] This specific embodiment adopts the following technical solutions: Its preparation process is: 1. Gene recombinant expression of hemagglutinin H7 antigen: (a), cloning of hemagglutinin H7 gene into a plasmid / host cell; (b), host Cell culture / plasmid replication; (c), host cells express H7 antigen; 2. Preparation of hemagglutinin H7 antigen: (d), host cell culture and amplification; (e), hemagglutinin pure protein antigen (F), pure hemagglutinin protein antigen and host cells; (g), use one-step and multi-step chromatographic methods for purification; (h), add stabilizer / adjuvant to make preparations; 3. use hemagglutination Pure protein antigen in animal / human body: (i), use hemagglutinin pure protein antigen to produce antibodies in animals; (j), the antibody against the hemagglutinin pure protein antigen has the effect of infection with viruses containing the hemagglutinin Protective effect; (k), use the hemagglutinin pure protein antigen to prevent infection of viruses...

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Abstract

The invention discloses a preparation method of a novel genetically recombinant pure protein antigen and relates to the technical field of protection of hemagglutinin viruses and development of detection reagents. The preparation process comprises the following steps: I, gene recombinant expression of a hemagglutinin H7 antigen; II, preparation of the hemagglutinin H7 antigen; III, application of the hemagglutinin pure protein to animals/human bodies; and IV, application of the high purity hemagglutinin H7 antigen to detecting hemagglutinin H7-containing viruses. The novel genetically recombinant pure protein antigen has the characteristics of high purity and good antigenicity, and does not exist in form of granules and not only does not contain any genetic materials such as RNA (Ribonucleic Acid), but also does not contain other protein impurities. The novel genetically recombinant pure protein antigen can be used for preparing a high specific antibody for H7N9. Because of the protein characteristic of the H7 antigen, the chromatographic method can be used for purifying a single antigen, and the purity of SEC-HPCL (Size Exclusion Chromatography-High Performance Liquid Chromatography) is higher than 90%. The high purity H7 antigen and the highly specific H7N9 antibody can be used for developing diagnostic reagents for diagnosing H7N9 avian influenza.

Description

Technical field [0001] The invention relates to the technical field of hemagglutinin virus protection and the development of detection reagents, in particular to a preparation process of a novel gene recombination pure protein antigen. Background technique [0002] H7N9 avian influenza is a new type of avian influenza, which was first discovered in Shanghai and Anhui at the end of March 2013. H7N9 avian influenza is the first new subtype of influenza virus discovered in the world. It has not yet been included in my country's legally reported infectious disease monitoring and reporting system, and no vaccine has been launched as of early April 2013. All patients infected by the virus have symptoms such as fever in the early stage. As of April 2013, it has not been confirmed whether the virus has the characteristics of human-to-human transmission. According to the investigation in April 2013, the H7N9 avian influenza virus gene was derived from genetic rematches between wild bird...

Claims

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Application Information

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IPC IPC(8): C12N15/63C07K14/11C07K16/10A61K39/145A61K39/42A61P31/16G01N33/68G01N33/569
Inventor 李晓辉
Owner SHANGHAI SIQI MEDICAL TECH
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