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Chimeric particle containing dominant epitope peptide of EB virus membrane surface glycoprotein gp350 and coding gene and application of chimeric particle

An Epstein-Barr virus and dominant epitope technology, applied in the field of genetic engineering, can solve the problems such as the lack of good treatment methods for cancer and the lack of Epstein-Barr virus preventive vaccines, and achieve the effect of improving immunogenicity

Active Publication Date: 2019-12-27
SUN YAT SEN UNIV CANCER CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Epstein-Barr virus is the first virus that has been confirmed to induce tumors after infection. More and more evidence shows that Epstein-Barr virus is associated with a variety of lymphoid tumors (including Hodgkin's lymphoma, Burkitt's lymphoma, NK / T cell lymphoma, etc.) etc.) and epithelial tumors (including nasopharyngeal carcinoma, gastric cancer, etc.) Preventive vaccines for viral infections are available, so it is particularly important to develop a vaccine against EBV infection

Method used

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  • Chimeric particle containing dominant epitope peptide of EB virus membrane surface glycoprotein gp350 and coding gene and application of chimeric particle
  • Chimeric particle containing dominant epitope peptide of EB virus membrane surface glycoprotein gp350 and coding gene and application of chimeric particle
  • Chimeric particle containing dominant epitope peptide of EB virus membrane surface glycoprotein gp350 and coding gene and application of chimeric particle

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1 Construction of recombinant expression vector and expression of fusion protein

[0033] 1. Experimental materials

[0034] (1) The present invention selects the hepatitis B core protein HBc149 as the granulated display carrier, transforms the carrier, replaces the 79-81 amino acids of HBc149 with "GGGGSGGGGTGSEFGGGGSGGGGS" and introduces BamH I and EcoR I double enzyme cleavage sites (GS / EF) , to facilitate the insertion of subsequent epitopes into recombinant expression vectors by enzyme-cut ligation.

[0035] (2) Reagents and consumables: All reagents and consumables are commercially available.

[0036] (3) Host cell: BL21DE3.

[0037] 2. Steps

[0038] (1) In order to fully investigate the ability of the dominant epitope to induce humoral immune response, the selected gp350 dominant epitope peptides were randomly combined and inserted into the granulated vector to construct an expression vector;

[0039] (2) Transform the above-mentioned vector into ...

Embodiment 2

[0054] Example 2 Immunoreactivity Verification of Recombinant Proteins

[0055] 1. Experimental materials

[0056] (1) Reagents and consumables: All reagents and consumables are commercially available.

[0057] (2) Antibody: HBc149-specific antibody 11H10 was a kind gift from Professor Xia Ningshao of Xiamen University; anti-gp350ECD123 multiple antiserum was prepared by immunizing mice in our laboratory.

[0058] 2. Experimental steps

[0059] (1) SDS-PAGE electrophoresis for the recombinant protein;

[0060] (2) After the electrophoresis is completed, the protein is transferred to the PVDF membrane;

[0061] (3) After the membrane transfer was completed, the PVDF membrane was blocked with 5% skimmed milk, and then the primary antibody and the secondary antibody labeled with horseradish peroxidase were incubated, and finally the substrate of horseradish peroxidase was used for color development.

[0062] 3. Experimental results

[0063] Such as image 3 As shown, the ex...

Embodiment 3

[0064] The granularity of embodiment 3 recombinant protein

[0065] 1. Experimental materials

[0066] (1) Reagents and consumables: All reagents and consumables are commercially available.

[0067] (2) Instruments: transmission electron microscope; Agilent HPLC analyzer; molecular sieve G5000PWxl.

[0068] 2. Experimental steps

[0069] (1) Staining the purified protein with tungsten phosphate;

[0070] (2) Transmission electron microscope detection after staining.

[0071] (3) After the purified protein is diluted to 0.5 mg / mL, the molecular sieve detection is carried out by using a high performance liquid chromatography analyzer.

[0072] 3. Experimental results

[0073] Such as Figure 4 As shown in A, the transmission electron microscopy test results show that all recombinant proteins have formed uniform particles; in addition Figure 4 B was detected by molecular sieves, and the results also showed that compared with the control protein wild-type HBc149, all recom...

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Abstract

The invention discloses a chimeric particle containing dominant epitope peptide of an EB virus membrane surface glycoprotein gp350 and a coding gene and application of the chimeric particle. Accordingto the chimeric particle and the coding gene and application thereof, it is found for the first time that the immunogenicity of polypeptide epitopes can be significantly improved by combining a plurality of dominant epitopes in a certain sequence and then conducting granulation display, immunized mice show that the total anti-titer of induced serum, the titer of neutralizing antibodies in the serum, and the efficiency of the serum in blocking viral infection at the cellular level are significantly improved. The invention provides a candidate vaccine form which can be used for developing and preventing EB virus infection, and important practical and theoretical significance and application prospects for prevention of EB virus-related diseases are achieved.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a chimeric particle comprising a dominant epitope peptide of the membrane surface glycoprotein gp350 of Epstein-Barr virus, its coding gene and its application. Background technique [0002] Epstein-Barr virus is widely infected in the global population, and it is reported that about 90% of adults have been infected with EBV. Infection with EBV in adolescence can easily lead to mononucleosis, which can usually be cured by itself; people who have been infected with EBV are generally life-long carriers of the virus. Epstein-Barr virus is the first virus that has been confirmed to induce tumors after infection. More and more evidence shows that Epstein-Barr virus is associated with a variety of lymphoid tumors (including Hodgkin's lymphoma, Burkitt's lymphoma, NK / T cell lymphoma, etc.) etc.) and epithelial tumors (including nasopharyngeal carcinoma, gastric cancer, etc....

Claims

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Application Information

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IPC IPC(8): C07K19/00C12N15/62A61K39/245A61P31/22
CPCA61K39/12A61P31/22C07K14/005C07K2319/00C12N2710/16222C12N2710/16234
Inventor 张晓曾益新曾木圣赵炳春徐淼冯启胜
Owner SUN YAT SEN UNIV CANCER CENT
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