Hepatitis B recombinant antigen, expression gene of hepatitis B recombinant antigen, construction method of hepatitis B recombinant antigen, virus-like particle of hepatitis B recombinant antigen, preparation method of virus-like particle of hepatitis B recombinant antigen, applications of hepatitis B recombinant antigen, and vaccine

A technology for recombining antigens and expressing genes, applied in the field of biomedicine, can solve the problems of poor immunogenicity and achieve good immunogenicity

Active Publication Date: 2017-07-07
PEKING UNIV SHENZHEN GRADUATE SCHOOL
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Problems solved by technology

[0005] Based on this, it is necessary to address the problem of poor immunogenicity of traditional hepatitis B virus vaccines, and provide a recombinant hepatitis B

Method used

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  • Hepatitis B recombinant antigen, expression gene of hepatitis B recombinant antigen, construction method of hepatitis B recombinant antigen, virus-like particle of hepatitis B recombinant antigen, preparation method of virus-like particle of hepatitis B recombinant antigen, applications of hepatitis B recombinant antigen, and vaccine
  • Hepatitis B recombinant antigen, expression gene of hepatitis B recombinant antigen, construction method of hepatitis B recombinant antigen, virus-like particle of hepatitis B recombinant antigen, preparation method of virus-like particle of hepatitis B recombinant antigen, applications of hepatitis B recombinant antigen, and vaccine
  • Hepatitis B recombinant antigen, expression gene of hepatitis B recombinant antigen, construction method of hepatitis B recombinant antigen, virus-like particle of hepatitis B recombinant antigen, preparation method of virus-like particle of hepatitis B recombinant antigen, applications of hepatitis B recombinant antigen, and vaccine

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preparation example Construction

[0082] For example, the present invention also provides a method for preparing preS VLP. The method for preparing preS VLP includes: inserting the base sequence of the expression gene of matrix protein M1 of influenza virus into the first expression vector to obtain the M1-expression vector plasmid, wherein, The base sequence of the expression gene of the matrix protein M1 of influenza virus is shown in SEQ ID NO.6; The preS-HA fusion gene as described in any one of the above examples is inserted into the second expression vector to obtain the preS-HA-expression vector plasmid ; The preS-HA-expression vector plasmid and the M1-expression vector plasmid are co-transfected into host cells, and the transfected host cells are cultured to produce preS VLP. For example, preS VLP is the hepatitis B recombinant antigen virus-like particle.

[0083] In order to enhance the release of influenza virus particles, in this embodiment, the 41st amino acid of influenza virus matrix protein M1...

Embodiment 1

[0099] 1. Plasmid construction

[0100] (1), construction of Pres-HA fusion gene

[0101] For the construction strategy of Pres-HA fusion gene, please refer to Figure 2A and Figure 2B .

[0102] The influenza virus hemagglutinin signal peptide region, the influenza virus transmembrane region and the intracellular region are obtained from the HA sequence of the influenza virus hemagglutinin. Wherein, the base sequence of the HA of the influenza virus hemagglutinin is shown in SEQ ID NO.8, the base sequence of the influenza virus hemagglutinin signal peptide region is shown in SEQ ID NO.3, and the influenza virus transmembrane region and cell The base sequence of the inner region is shown in SEQ ID NO.4. The influenza virus hemagglutinin signal peptide region is a DNA molecule encoding the 1st to 20th amino acids of the influenza virus hemagglutinin, and the 1st to 20th amino acid sequence of the influenza virus hemagglutinin is shown in SEQ ID NO.9. The transmembrane reg...

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Abstract

The invention relates to a hepatitis B recombinant antigen, an expression gene of the hepatitis B recombinant antigen, a construction method of the hepatitis B recombinant antigen, a virus-like particle of the hepatitis B recombinant antigen, a preparation method of the virus-like particle of the hepatitis B recombinant antigen, applications of the hepatitis B recombinant antigen, and a vaccine. The hepatitis B recombinant antigen is obtained by expressing a polynucleotide comprising a nucleotide sequence shown in the SEQ ID NO.1. The expression gene of the hepatitis B recombinant antigen is obtained by splicing a preS region of the hepatitis B virus with a signal peptide region of an influenza virus hemagglutinin and splicing a transmembrane region of the influenza virus with an intracellular region, the expression gene is successfully expressed to obtain the hepatitis B recombinant antigen, and the hepatitis B recombinant antigen is better in immunogenicity, and can enable the immune system of the minority (about 5%-10%) incapable of producing antibodies for the traditional vaccine to make an immune response. Meanwhile, on the basis of the indispensable importance of the preS region for the hepatitis B virus, the vaccine for broader intended population is prepared for the preS region. The vaccine has the effects of preventing infection of the hepatitis B virus and clearing away the hepatitis B virus, and is expected to be transformed into a novel preventive and therapeutic hepatitis B vaccine on medical science.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a hepatitis B recombinant antigen and its expression gene, a construction method, a virus-like particle and its preparation method, application and vaccine. Background technique [0002] Hepatitis B is one of the major infectious diseases of human beings, affecting as many as 300 million people worldwide. According to the survey, about one tenth of the people in China are carriers of hepatitis B virus. Hepatitis B virus infection can cause acute and chronic hepatitis in humans, and hepatitis can develop into more serious diseases, such as liver cirrhosis and liver cancer. [0003] Hepatitis B Virus (HBV), referred to as HBV, belongs to the hepadnaviridae family and is susceptible to humans and chimpanzees. There are three kinds of antigen molecules on the surface of hepatitis B virus particles, which are the small protein (the small protein, S protein), the middle protein (...

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62C12N7/04C12N15/85A61K39/29A61P31/20A61P1/16
Inventor 王国新
Owner PEKING UNIV SHENZHEN GRADUATE SCHOOL
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