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A fusion antigen of the first hypervariable region of hepatitis C virus and its application

A hepatitis C virus, fusion antigen technology, applied in the direction of hybrid peptides, material testing products, instruments, etc., can solve problems such as the inability to meet the activity requirements, and achieve the effect of eliminating missed detection, simple operation and good repeatability

Active Publication Date: 2017-04-12
山东莱博生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

This is obviously not able to meet the activity requirements of the coating antigen in the diagnosis of HCV

Method used

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  • A fusion antigen of the first hypervariable region of hepatitis C virus and its application
  • A fusion antigen of the first hypervariable region of hepatitis C virus and its application
  • A fusion antigen of the first hypervariable region of hepatitis C virus and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Example 1: Selection of Candidate Antigen Epitopes

[0036] The applicant searched the HCV sequences database (http: / / hepatitis.ibcp.fr.), analyzed a total of 1,700 HVR1 sequences of 6 genotypes of HCV, and selected 6 sequences according to the homology of amino acid sequences. HVR1 antigen sequences of 6 genotypes of HCV. Wherein, the amino acid sequences of the first hypervariable region (HVR1) antigens of the six hepatitis C virus HCV1-HCV6 genotypes are as follows:

[0037] ①STYATGGAAGRETSTWAGLFNPGAAQK

[0038] ②TTYTTAGSAARATYTLTSLFKTGPKQK

[0039] ③ETHVTGGTAAHGALGITSLLSRGPKQN

[0040] ④ GTHVTGGKAG HTIHGFTSLFTRGSAQN

[0041] ⑤STHVTGGVQGRTTRGLTSLFTPGPPSQK

[0042] ⑥ TTY VSGGASG FNTHVFTGIF SSGASQK.

Embodiment 2

[0043] Example 2: Cloning and expression of multitype HVR1 fusion antigen, that is, the first hypervariable region fusion antigen of hepatitis C virus

[0044] 1. Construction of multitype HVR1 fusion antigen expression plasmid

[0045] 1.1 Synthesis of multitype HVR1 fusion antigen and gene sequence synthesis of multitype HVR1 fusion antigen

[0046] According to the amino acid sequences of the 6 HVR1 antigens belonging to the 6 genotypes of HCV described in Example 1, Shanghai Jierui Bioengineering Co., Ltd. was entrusted to synthesize the HVR1 polypeptides of each genotype.

[0047] Utilize gene recombination method, add flexible short peptide (GGGGS or GGGGSGGGGSGGGGS or AAY, preferably GGGGSGGGGSGGGGS) to the HVR1 antigen amino acid sequence that belongs to 6 genotypes of HCV 6 described in Example 1 and make it link together (its amino acid sequence is as SEQ ID No.1), using Escherichia coli preferred codons, translated the amino acid sequence into nucleotide sequence, ...

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Abstract

The invention discloses a fusion antigen of the first hypervariable region of hepatitis C virus. The amino acid sequence of the fusion antigen is shown in SEQ ID No.1. The invention also discloses the detection of the fusion antigen in the preparation of hepatitis C virus HVR1 antibody. application in the kit. It is verified by experiments that the cross-reactivity of the fusion antigen of the present invention is significantly improved compared with the single fragment HVR1 antigen, and the detection coincidence rate with hepatitis C virus RNA-positive patients is as high as 99%. Since HVR1 is the main neutralizing epitope in hepatitis C virus, through the application of the antigen of the present invention, for the detection of hepatitis C virus HVR1 antibody, the purpose of comprehensive detection of HCV infection can be realized, and it can be used for the transformation of HCV disease. Problems such as reconciliation prognosis and interferon curative effect prediction can be effectively solved, and have great clinical application prospects.

Description

technical field [0001] The invention relates to a hepatitis C virus antigen, in particular to a fusion antigen of the first hypervariable region (HVR1) of the hepatitis C virus and its application in the preparation of a detection kit for the hepatitis C virus HVR1 antibody. Background technique [0002] The most notable clinical feature of hepatitis C virus (hepatitis C virus, HCV) infection is that it causes chronic infection. About 70% of hepatitis C may develop into chronic hepatitis, 20% into liver cirrhosis, and 12% into liver cancer, which is very harmful. seriously. There are about 170 million people in the world carrying HCV, and it is estimated that there are 43 million HCV-infected people in my country, and the number of new cases is rising every year, and the infection rate is about 3.2%. [0003] Since the HCV gene was cloned in 1989, people have been working on vaccine research, but because the viral RNA is low in blood and very unstable, the HCV genome mutati...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00G01N33/68G01N33/576
Inventor 续薇陈振丁兆明王岩欧兰香王佳颖寇宗阳丁兴龙王恒朱之炜
Owner 山东莱博生物科技有限公司
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