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Subunit vaccine based on prokaryotic expression of norovirus antigenic epitope and preparation method of subunit vaccine

A subunit vaccine and virus antigen technology, applied in the field of biomedicine, can solve the problems that intestinal-like tissues are difficult to establish and maintain, and are not suitable for the development and production of large-scale vaccines, and achieve high sensitivity, simple operation, and specificity strong effect

Active Publication Date: 2019-04-09
INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The in vitro culture of Norovirus has always been an important factor restricting Norovirus vaccines. At the same time, the lack of suitable animal models is also the key to restricting the development of traditional vaccines. Although the latest research shows that intestinal-like tissues derived from human small intestinal stem cells can Norovirus can be cultivated in vitro, but intestinal-like tissues are difficult to establish and maintain, and are not suitable for traditional large-scale vaccine development and production. Therefore, genetic engineering vaccines have become the best choice for norovirus vaccine development

Method used

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  • Subunit vaccine based on prokaryotic expression of norovirus antigenic epitope and preparation method of subunit vaccine
  • Subunit vaccine based on prokaryotic expression of norovirus antigenic epitope and preparation method of subunit vaccine
  • Subunit vaccine based on prokaryotic expression of norovirus antigenic epitope and preparation method of subunit vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0023] 1 material

[0024] 1.1 Norovirus recombinant antigen sequence design

[0025] Source: NCBI Reference Sequence: NC_029646.1

[0026] Neutralizing epitope of human norovirus type GII: 105bp.

[0027] The specific sequence is as follows:

[0028] GGATCCAACCACGTGTGGAACATGCAGGTT GGTGGCGGTGGCAGC GCGAAGTTCACCCCGAAACTGGGTGCGATCCAAATTGGCACCTGGGAGGAAGACGAATTC The underlined part is the link peptide

[0029] 1.2 Enzyme

[0030] name

Remark

BamHI

Purchased from NEB Corporation

EcoRI

Purchased from NEB Corporation

T4 ligase

Purchased from NEB Corporation

[0031] 1.3 Carrier

[0032]

[0033] 1.4 Reagents and kits

[0034]

[0035]

[0036] 1.5 Instruments

[0037] name

Remark

shaker

Purchased from Beijing Liuyi Instrument Factory, model WD-9405B

Electrophoresis

Purchased from US BIO-RAD company, the model is 200 / 2.0Power Supply

Sonicator

Purchased from Ningbo Xinzhi C...

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Abstract

The invention relates to the field of biomedicine, in particular to a preparation method of a subunit vaccine based on prokaryotic expression of a norovirus antigenic epitope. The invention provides the prokaryotic expressed subunit vaccine which is used for stimulating a body to produce a specific antibody against norovirus, and is characterized in that the norovirus recombinant antigen sequenceis as shown in SEQ ID NO: 1, source: NCBI Reference Sequence: NC_029646.1, a neutralizing epitope of human norovirus GII type: 105 bp, and an expression vector: a modified hepatitis B core antigen (HBcAg) gene cloned into a pThioHisA expression vector.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a preparation method based on prokaryotic expression of norovirus epitope subunit vaccines. Background technique [0002] Norovirus (NoV) is the main pathogen of human acute gastroenteritis, which was called "winter vomiting disease" in 1929 because of the obvious seasonality of gastroenteritis caused by it. The complexity of norovirus genotypes and the rapid mutation between norovirus types make the development of norovirus vaccines slow. So far, there is no stable and effective vaccine for mass vaccination, nor is there a safe and stable anti-virus vaccine. Venom serum is used for emergency protection of high-risk exposed groups. [0003] Norovirus belongs to the Norwalk virus genus of the family Calicivirus, and the viral RNA contains three open reading frames (ORFs). Noroviruses can be divided into seven genomes according to their RNA polymerase and capsid protein sequence homolo...

Claims

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Application Information

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IPC IPC(8): A61K39/12C07K14/08C07K1/36C07K1/30C07K1/18A61P31/14
CPCA61K39/12A61P31/14C07K14/005C12N2770/10051C12N2770/16034C12N2770/16022Y02A50/30
Inventor 李鸿钧刘洋林晓晨周艳吴晋元解裕萍
Owner INST OF MEDICAL BIOLOGY CHINESE ACAD OF MEDICAL SCI
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