Mosaic type virus-like particle DNA vaccine

A DNA vaccine and virus-like technology, applied in the field of DNA vaccines, can solve the problems of complex preparation and purification process and high cost, and achieve the effect of protecting against pathogen infection, simple preparation and improved ability

Inactive Publication Date: 2007-01-31
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] However, the above modification is only to purify the product expressed in vitro and then immunize the body to generate an immune response.
Its preparation and purification process is complex and costly

Method used

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  • Mosaic type virus-like particle DNA vaccine
  • Mosaic type virus-like particle DNA vaccine
  • Mosaic type virus-like particle DNA vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1, the preparation of chimeric virus-like particle DNA vaccine

[0039] 1. Construction of the recombinant expression vector proVAX-ABC containing the gene encoding the chimeric protein HBc-VP1

[0040] In order to enable the foot-and-mouth disease virus VP1 gene to be effectively expressed in the human hepatitis B virus core antigen and to be displayed outside the core antigen, the foot-and-mouth disease virus VP1 gene fragment was cloned into the 78th amino acid and the 80th amino acid from the amino terminal of the human hepatitis B virus core antigen. between the codons of the amino acids to facilitate the display and presentation of the antigen. The specific method is as follows:

[0041] (1) Primer design: according to the sequences of human hepatitis B virus core antigen gene and foot-and-mouth disease virus VP1 gene, and according to figure 1 3 sets of primers were designed according to the cloning ideas in . The primer sequences are

[0042] CP01...

Embodiment 2

[0056] Example 2, Animal experiment of chimeric virus-like particle DNA vaccine proVAX-ABC

[0057] 1. Mouse experiment

[0058] 4-6 weeks old female C57BL / 6 mice (purchased from the Institute of Experimental Animals, Chinese Academy of Medical Sciences), divided into 4 groups, 6 mice in each group, were immune to proVAX-VP1 (100 μg / mouse / time), proVAX-ABC (100μg / one / time), proVAX (100μg / one / time) and its PBS control group (100μl / one / time). Each mouse was first immunized on day 0 and boosted once on day 14. Mouse serum was collected 14, 28, 42, 56, and 70 days after the first immunization, and stored at -20°C for future use.

[0059] (1) ELISA detection of IgG antibody levels in proVAX-VP1 immunized mice

[0060] Coat 96-well ELISA plate with 1 μg / ml 146S antigen (remove the mineral oil in the bovine foot-and-mouth disease O-type inactivated vaccine (purchased from Inner Mongolia Jinyu Biopharmaceutical Factory) to obtain 146S antigen), 100 μl / well, overnight at 4°C; PBST ...

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Abstract

A chimeric virus-like granular DNA vaccine able to generate stronger B cell activity, exciting high antibody level, improving cell's immunizing level by cross presentation, and activating T cell, toxic T cell and complete immunoreaction is a recombinant eucaryotic expression carrier of coding gene with chimeric protein. Its preparing process is also disclosed.

Description

technical field [0001] The invention relates to a DNA vaccine, in particular to a chimeric virus-like particle DNA vaccine. Background technique [0002] There are many ways to prevent infection by various pathogens, and improving the body's immunity is the most important means, which is generally achieved by vaccination. Vaccination is an effective measure to prevent infection by various pathogens. Common pathogens are organisms and molecules such as viruses, microorganisms, eukaryotic cells, parasites, and environmental agents. Various methods have been used to produce vaccines against infectious pathogens, such as inactivated vaccines, live attenuated vaccines, recombinant vaccines, subunit vaccines and nucleic acid vaccines. Their basic principles of action are the same, that is, to stimulate immune responses with the help of histocompatibility proteins combined with pathogens, so as to prevent immune individuals from being infected by infectious pathogens. When an in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/295A61K48/00A61K39/39A61P31/12C12N15/85C12N15/51
Inventor 王宾金华利
Owner CHINA AGRI UNIV
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