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Varicella-zoster virus gE protein mutant and expression method thereof

A herpes zoster virus and mutant technology, which is applied to viruses/bacteriophages, viruses, viral peptides, etc., can solve problems such as unsatisfactory immune effect, low protein activity, and low VZVgE protein expression efficiency

Active Publication Date: 2020-12-29
IMMUNE PATH BIOTECHNOLOGY SUZHOU CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Although some vaccines against VZV have been developed in the prior art, there are still problems such as low expression efficiency of VZV gE protein, low activity of the expressed protein, and unsatisfactory immune effect.

Method used

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  • Varicella-zoster virus gE protein mutant and expression method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Cloning construction, expression and purification of varicella-zoster virus gE protein

[0034] 1. Selection of gE protein and synthesis (of gE gene)

[0035] Through NCBI database and literature search, a conservative truncated gE protein amino acid sequence was selected as the basis for gene optimization. In order to improve expression efficiency, only the signal peptide region and mature antigen were selected. At the same time, the inventors of the present application surprisingly found that when selecting a specific amino acid sequence as the basis for gene optimization in the process of preparing recombinant gE protein, if the 141st position of the mature antigen region is artificially modified (mutated from leucine to methionine acid), the genes and vectors designed based on the modified amino acid sequence will be able to achieve higher antigen expression. The full length of the mutated protein sequence is 546 amino acids (SEQ ID NO: 1), and the detail...

Embodiment 2

[0058] Example 2 Immunogenicity Evaluation of Recombinant Herpes Zoster Vaccine

[0059] Obtain the gE protein described in Example 1, and after conventional processing methods, such as hydrophobic chromatography, anion exchange chromatography, hydroxyapatite chromatography, ultrafiltration and nanofiltration, the protein with a purity of more than 95% can be obtained used as an antigenic protein.

[0060] In order to study the immunogenicity of the antigen prepared by the gene and carrier provided by the present invention, the above-mentioned antigen and adjuvant were sucked and formulated into a recombinant herpes zoster vaccine composition, and the C57BL / 6 mouse was used as an animal model to carry out immunization Original research. The specific method is as follows: the gE protein is used as an antigen, and aluminum phosphate and CpG ODN are used as adjuvants to inhale and prepare the recombinant herpes zoster vaccine composition. Select 6-8 week-old C57BL / 6 mice into r...

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Abstract

The invention relates to a varicella zoster virus gE protein mutant and an expression method thereof. Specifically, the invention discloses a protein with immunogenicity and a coding gene thereof. Theinvention also discloses a preparation method of the protein with immunogenicity.

Description

technical field [0001] The invention belongs to the field of biomedicine, in particular, the invention relates to a recombinant herpes zoster vaccine composition and its preparation method and application. Background technique [0002] Varicella zoster virus (VZV) is one of eight human herpesviruses, also known as human herpesvirus type 3. Varicella-zoster virus spreads all over the world and is highly contagious. So far only one serotype has been found, and VZV only infects humans in nature. It can cause both chickenpox and herpes zoster (herpes zoster, HZ). Chickenpox is usually seen in childhood, and shingles will not appear until adulthood. After primary infection with varicella, the virus can remain latent in the ganglion of the host, and with age, immunocompromise, or immunosuppression, VZV can be reactivated and cause herpes zoster. Herpes zoster presents clinically as a unilateral vesicular eruption, markedly limited to a single skin segment, often with radicular p...

Claims

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Application Information

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IPC IPC(8): C07K14/04C12N15/38C12N15/85C12N5/10
CPCC07K14/005C12N15/85C12N5/0682C12N2710/16722C12N2800/107C12N2510/02C07K14/04C12N5/10C12P21/02C12N15/63
Inventor 周晨亮何强沈巧英周凌云江元翔刘革曾宪放史力莫呈钧张智
Owner IMMUNE PATH BIOTECHNOLOGY SUZHOU CO LTD
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