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Epitope of Epstein-Barr virus latent membrane protein 2b and its application

A latent membrane protein and Epstein-Barr virus technology, applied in antiviral immunoglobulin, application, antiviral agents, etc., can solve the problem of weak immunogenicity

Inactive Publication Date: 2011-12-28
WENZHOU MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

②Synthetic peptide vaccine: Studies have proved that LMP2 is the main target antigen for the body to generate CTL responses and anti-tumor effects. Selecting its CTL epitopes and artificially synthesizing peptide chains can produce certain protective cellular immunity, but the immunogenicity is weak
④ EBV antigen-loaded dendritic cell (DC) vaccine: Transfect DC with adenovirus vector cloned from EBV LMP2 gene, or extract B cell protein infected by EBV, induce DC as a vaccine, and the immune protection effect induced by it is still in progress under study

Method used

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  • Epitope of Epstein-Barr virus latent membrane protein 2b and its application
  • Epitope of Epstein-Barr virus latent membrane protein 2b and its application
  • Epitope of Epstein-Barr virus latent membrane protein 2b and its application

Examples

Experimental program
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Effect test

preparation example Construction

[0096] The present invention also provides a kit (kit) for detecting diseases related to Epstein-Barr virus infection. The kit contains: a solid phase carrier coated with the B cell surface bit recombinant protein. The preparation method of the kit (kit) includes: (1) coating the B cell epitope recombinant protein on a solid phase carrier (such as an ELISA reaction plate), and obtaining the B cell epitope coated and (2) placing the solid phase carrier coated with the B cell epitope recombinant protein obtained in (1) into a kit, thereby obtaining a drug for detecting Epstein-Barr virus infection-related diseases box. The kit may also include reagents (such as enzyme-linked immunosorbent reagents) for detecting antigen-antibody reactions, or reagents for gene amplification (such as PCR reagents) in appropriate containers, and / or also Instructions for use (book) are included.

[0097] Detection purpose

[0098] The B cell epitope recombinant protein of the invention can be u...

Embodiment 1

[0108] Example 1, Preparation and Identification of EBV-LMP2B Cell Epitope Recombinant Protein

[0109] 1. Design of EBV-LMP2B cell epitope recombinant protein gene

[0110] Obtain the EBV latent membrane protein 2 (LMP2) gene sequence and amino acid sequence from the network resource database (Genbank, Swiss-Prot); use the online network resource (EXPASY) and the biological software DNASTAR to analyze the antigenicity and surface accessibility of the EBV-LMP2 amino acid sequence The parameters such as property, flexibility and transmembrane region were analyzed ( figure 1 ), supplemented by the analysis results of the secondary structure of EBV-LMP2 protein, analysis and comparison of various parameters to comprehensively judge the B cell dominant epitope of EBV-LMP2.

[0111] After the above-mentioned epitope prediction, combined with repeated tests and selections by the inventors, the results obtained three amino acid sequences derived from EBV latent membrane protein 2, w...

Embodiment 2

[0117] Example 2. Immunogenicity Study of EBV-LMP2B Cell Epitope Recombinant Protein

[0118] Female BALB / c mice aged 6-8 weeks were randomly divided into 5 groups, with 9 mice in each group: Groups 1-3 were the EBV-LMP2B cell epitope protein immunization group, and group 4 was the His-tag protein control group . B cell epitope protein and Freund's adjuvant (FCA) 1:1 (W / W) were fully emulsified evenly, and at 0, 2, and 4 weeks, 50 μg of B cell epitope protein or His-tag protein per mouse, Mice were immunized subcutaneously on the back. The humoral immune effect of the immunized mice was detected, that is, the titer and maintenance time of specific serum IgG.

[0119] At 0, 3, and 6 weeks, each group of mice was blooded by docking the tail to detect the level of serum IgG antibody by ELISA method.

[0120] 1. Determination results of serum-specific IgG antibodies of immunized mice

[0121] The three B cell epitope proteins of purified EBV-LMP2 were coated on the ELISA react...

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Abstract

The invention relates to the preparation and application of EB virus latent membrane protein 2B cell epitope recombinant protein. The invention discloses three B cell epitopes screened on the basis of full-length Epstein-Barr virus latent membrane protein 2. The invention also discloses the coding nucleotide and amino acid sequences of the B cell epitope. The invention also discloses the use of the B cell epitope in preventing, treating and diagnosing Epstein-Barr virus infection and related diseases. The protein of the invention has strong immunogenicity and antigenicity, and has good application prospect.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals and diagnosis, more specifically, the present invention relates to three B cell epitopes from latent membrane protein 2 of Epstein-Barr virus and its encoding nucleic acid, its preparation method and its application in the prevention, treatment and diagnosis of Epstein-Barr virus Applications in infection and related diseases. Background technique [0002] Epstein-Barr Virus (EBV), the pathogen of infectious mononucleosis (IM), was first discovered by Epstein and Barr in 1964 from cultured cells of Burkitt's lymphoma in Africa. EBV infection is common, and more than 90% of people have established lifelong latent infection (see Maeda A, Sato T, Wakiguchi H. [Epidemiology of Epstein-Barr virus (EBV) infection and EBV-associated diseases] [J]. Nippon Rinsho, 2006, 64Suppl 3:609-612), but there were no obvious clinical symptoms. EBV has the characteristics of B lymphocytes and can stimulate the pro...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K16/08C12N15/38C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10C12P21/02C12Q1/70C12Q1/68G01N33/574G01N33/569G01N33/543A61K39/245A61K48/00A61P31/20A61P35/00A61P35/02
Inventor 张丽芳朱珊丽薛向阳李文姝文金生
Owner WENZHOU MEDICAL UNIV
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