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Multi-epitope recombinant protein of epstein-barr (EB) virus latent membrane protein 2 and application thereof

A latent membrane protein, Epstein-Barr virus technology, applied in the fields of application, viral peptides, antiviral agents, etc., can solve the problems of difficult to reach a large sensitive population, weak immunogenicity, etc.

Inactive Publication Date: 2011-03-30
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
In conclusion, research on CTL-based vaccines shows attractive prospects for the treatment of EBV infection and related tumors, but the problem of weak immunogenicity has yet to be resolved. Purpose of Sensitive Groups

Method used

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  • Multi-epitope recombinant protein of epstein-barr (EB) virus latent membrane protein 2 and application thereof
  • Multi-epitope recombinant protein of epstein-barr (EB) virus latent membrane protein 2 and application thereof
  • Multi-epitope recombinant protein of epstein-barr (EB) virus latent membrane protein 2 and application thereof

Examples

Experimental program
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Effect test

preparation example Construction

[0094] 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 multi-epitope Recombinant protein. The preparation method of the kit (kit) comprises: (1) coating the multi-epitope recombinant protein on a solid-phase carrier (such as an ELISA reaction plate) to obtain the multi-epitope recombinant protein coated and (2) placing the solid phase carrier coated with the multi-epitope recombinant protein obtained in (1) into a kit, so as to obtain a kit for detecting diseases related to Epstein-Barr virus infection. 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.

[0095] Detection purpose

[0096] The multi-epitope recombinant protein of the invention can be...

Embodiment 1

[0106] Example 1, Preparation and identification of EBV-LMP2 multi-epitope recombinant protein

[0107] 1. Design of EBV-LMP2 multi-epitope recombinant protein gene

[0108] Apply the network resource database (Genbank, Swiss-Prot) EBV latent membrane protein 2 (LMP2) gene and amino acid sequence; according to the most common HLA genes in the Chinese Han population are HLA-A*02, HLA-A*24, HLA-B* 58 and HLA-DRB 1*15, HLA-DRB 1*03 genes (Zeng Xuehui, Xiao Lulu, Li Jiang et al.; Study on the correlation between HLA-A, B, DRB1 allelic polymorphisms and nasopharyngeal carcinoma in southern China[J ]. Journal of Cellular and Molecular Immunology, 2007 (9): 819-821; Song Yonghong, Ma Chunhong, Lu Hongjuan, etc.; Study on HLA gene polymorphisms of the Han population in northern China [J]. Shandong University Journal (Medical Edition), 2007 (6 ): 546-553.), using the online software (SYFPEITHI, EXPASY) and the biological software DNASTAR to predict the above-mentioned HLA gene-restric...

Embodiment 2

[0129] Example 2, Immunogenicity Research of EBV-LMP2 Multi-epitope Recombinant Protein

[0130] Female BALB / c mice aged 6-8 weeks were randomly divided into 3 groups, 9 mice in each group: the first group was the EBV-LMP2 multi-epitope protein immunization group, and the second group was the pET32a(+) empty vector control group; Group 3 was the PBS blank control group. Multi-epitope protein and Freund's adjuvant (FCA) 1:1 (W / W) were fully emulsified evenly, and at 0, 2, and 4 weeks respectively, 50 μg of multi-epitope protein or empty carrier protein was added to the back of each mouse. Spot immunize mice. The effects of humoral immunity and cellular immunity were tested on the immunized mice, that is, the titer and maintenance time of specific serum IgG and vaginal secretion sIgA, and the specificity of CTL was detected by lactate dehydrogenase (LDH) release method for cellular immunity lethal activity.

[0131] At 0, 1, 3, 5, and 7 weeks, the blood of each group of mice ...

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Abstract

The invention relates to preparation and application of multi-epitope recombinant protein of epstein-barr (EB) virus latent membrane protein 2. The invention discloses the multi-epitope recombinant protein rich in a plurality of CTL epitopes, Th epitopes and B cell epitopes obtained by screening based on the full-length EB virus latent membrane protein 2. The invention also discloses the coding nucleic acid of the protein, and comprises a nucleic acid recombinant vector and a host cell. The invention also discloses the application of the protein in the aspects of preventing, treating and diagnosing EB virus infection and related disease thereof. The protein of the invention has very strong immunogenicity and antigenicity and good application prospect.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals and diagnosis, more specifically, the invention relates to a multi-epitope recombinant protein composed of a plurality of CTL, Th and B cell epitopes on the latent membrane protein 2 of Epstein-Barr virus, its encoding nucleic acid, and its preparation The method and its application in the prevention, treatment and diagnosis of Epstein-Barr virus infection and related diseases. Background technique [0002] Epstein-Barr Virus (EBV), the pathogen of infectious mononucleosis, 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 no obvious clinical symptoms. EBV has the characteristics of B ...

Claims

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Application Information

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IPC IPC(8): C07K14/05C12N15/38C12N15/63C12N1/21C12N1/19C12N5/10C12P21/02A61K39/245A61K48/00A61P31/22A61P35/00G01N33/569C12Q1/68A61J1/00C12R1/93
Inventor 张丽芳薛向阳朱珊丽陈韶陆丽君
Owner WENZHOU MEDICAL UNIV
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