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Virus protein granules of hepatitis E virus-4 ORF2 fragment and prepration method and application thereof

A technology of hepatitis E virus and protein particles, which is applied in the field of protein particles of hepatitis E virus, and can solve the problem that the specific function of the protein is not clear

Inactive Publication Date: 2010-11-10
NAT INST FOR THE CONTROL OF PHARMA & BIOLOGICAL PROD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Among them, the product encoded by ORF1 is a viral functional protein; ORF2 encodes a glycosylated protein containing 660 amino acids, with a molecular weight of about 72kD, which is a viral capsid protein, which contains at least 7 linear epitopes, especially its C-terminus. An important immune epitope; ORF3 partially overlaps with ORF1 and ORF2, and its encoded protein contains at least 4 linear epitopes, but the specific function of this protein is still unclear

Method used

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  • Virus protein granules of hepatitis E virus-4 ORF2 fragment and prepration method and application thereof
  • Virus protein granules of hepatitis E virus-4 ORF2 fragment and prepration method and application thereof
  • Virus protein granules of hepatitis E virus-4 ORF2 fragment and prepration method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0207] Example 1 Expression of Hepatitis E Virus Protein Particles in Insect Baculovirus Expression System

[0208] 1. Culture of Insect Cells

[0209] (1) Cryopreservation of Sf9 cells

[0210] cell line

Freezing medium

Density (cell / ml)

Adhesion time after resuscitation (min)

Sf9

60% Grace's Insect Medium 30% FBS 10% DMSO

1×10 7

3045

[0211] a. Count the cells with a cell counter.

[0212] b. Place the sterile cell cryopreservation tube on ice and mark it.

[0213] c. Centrifuge at 400-600g room temperature for 10min, discard the supernatant.

[0214] d. Resuspend the cells at the above density with the above freezing medium.

[0215] e. Transfer 1ml of the cell suspension to a sterile cell cryovial.

[0216] f. Store at -20°C for 1 hour, and at -80°C for 24-48 hours.

[0217] g. Freeze in liquid nitrogen.

[0218] (2) Recovery and adherent culture of Sf9 cells

[0219] a. Place the cryopreservation tube in warm wat...

Embodiment 2

[0386] Through the experimental procedure of Example 1, the amino acid sequence of the sequence shown in SEQ NO: 1, in which positions 1-21 are the N-terminus of the protein and positions 497-516 are the C-terminus of the protein, can obtain viral protein particles similar to those in Example 1.

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Abstract

The invention relates to protein granules of hepatitis E virus, in particular to virus protein granules of a hepatitis E virus-4 ORF2 fragment, recombinant expression vectors, host cells and antibodies of hepatitis virus protein granules, application and expression methods of the hepatitis virus protein granules, the antibodies and nucleotide sequences, and preparation methods and application thereof. The sequences of the virus protein granules have at least 477 continuous nucleotide sequences in the sequences expressed as SEQ No: 1 or SEQ No: 3.

Description

technical field [0001] The invention relates to a protein particle of hepatitis E virus, in particular to a virus protein particle of the hepatitis E virus type 4 ORF2 fragment, as well as its preparation method and application. Background technique [0002] The pathogen of hepatitis E is hepatitis E virus (hepatitis E virus, HEV), which is a single-stranded positive-sense RNA virus without envelope, and only one serotype has been found so far. Hepatitis E is mainly transmitted through the fecal-oral route. In addition, there is evidence that HEV can be transmitted through non-fecal-oral routes and mother-to-child transmission in addition to intestinal transmission in hepatitis E-endemic areas. Xia et al. took 10ml of human venous blood with HEV in the body and injected it into rhesus monkeys, and found that HEV existed in the monkeys, indicating that blood is also a dangerous way of HEV transmission. In addition to causing acute hepatitis E, more than 50% of acute liver fa...

Claims

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Application Information

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IPC IPC(8): C07K14/08C12N15/51C07K16/10C12N15/63C12N5/10C12P21/02C12N15/85A61K39/29A61K39/42A61P31/14G01N33/576C12Q1/70C12R1/93
Inventor 王佑春樊金萍黄维金赵晨燕
Owner NAT INST FOR THE CONTROL OF PHARMA & BIOLOGICAL PROD
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