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Hepatitis B virus surface L protein related peptide

A protein and virus technology, applied in viral peptides, antiviral agents, peptide/protein components, etc.

Active Publication Date: 2006-02-15
SHANGHAI HEP PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 2. Current status of prevention and treatment of HBV infection
In addition, other adverse reactions such as antigen-antibody complex immunopathology may also occur

Method used

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  • Hepatitis B virus surface L protein related peptide
  • Hepatitis B virus surface L protein related peptide
  • Hepatitis B virus surface L protein related peptide

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1: Inhibition of HBV hepatocyte infection by peptides derived from HBV surface L protein of B genotype adw serotype and C genotype adr serotype

[0085] 1. Preparation of peptides derived from surface L protein of HBV serotype B genotype adw serotype and C genotype adr serotype

[0086] N-terminal myristoyl-modified HBV surface L protein-derived peptides of B genotype adw serotype and C genotype adr serotype (SEQ ID NO: 3 amino acid glycine modified with myristoyl, expressed as Myr 13-59) The AB 431A peptide synthesizer follows the standard Fmoc protocol and uses 0.25mM HMP as the starting resin to extend the synthesis residue by residue from the carboxyl end to the amino end. To enhance the stability of the polypeptide, the C-terminus of the polypeptide was further amidated. After peptide synthesis, cut with cutting solution, filter out resin with G6 glass sand funnel, and vacuum dry the filtrate. The peptide cleavage product was dissolved in ion-free water, ...

Embodiment 2

[0096] Example Two: Inhibition of HBV Hepatocyte Infection by Peptides Derived from Surface L Protein of D Genotype Ayw Serotype HBV Virus

[0097] 1. The synthesis of Myr 2-47 (SEQ ID NO: 9) derived from surface L protein of HBV serotype D ayw serotype D with myristoyl modification (SEQ ID NO: 9) and the culture of primary human hepatocytes are the same as in Example 1.

[0098] 2. Cultivation of D genotype ayw serotype HBV virus

[0099] The 2.2.15 cell line, which can secrete complete D genotype ayw serotype HBV infected virus particles, was continuously cultured for 10 days. The culture supernatant was collected, centrifuged with 6% polyethylene glycol (PEG8000) to precipitate virus particles, the pellet was resuspended in phosphate buffer containing 25% fetal calf serum, and frozen at -80°C.

[0100] 3. Detection of HBsAg in hepatocyte culture supernatant after HBV infection (same as embodiment one)

[0101] 4. Inhibition of HBV hepatocyte infection by HBV surface L pro...

Embodiment 3

[0103] Example 3: Screening of HBV surface L protein derivatized peptides

[0104] 1, B genotype adw serotype and C genotype adr serotype HBV virus genotype serotype determination, virus culture, cultivation of human primary hepatocytes (same example 1).

[0105] 2. The construction of a recombinant plasmid carrying a complete HBV genome and having the complete replication ability of HBV.

[0106] Add 310 μl proteinase K lysate to 50 μl HBV virus culture suspension [1mg / ml proteinase K, 50mmol / L Tris-HCl (pH8.0), 200mmol / L NaCl, 10mmol / L EDTA, 2% SDS, 1μg / ml poly(A )]. After lysis at 60°C for 1 hour, extract with phenol / chloroform, precipitate with ethanol, and dissolve the precipitate in H 2 O is HBV DNA solution. Entire EN II replication element was amplified with upstream primer (Pst I) 5'ctgactgcagCACTGGATGGGGCTTGGCTATTGG (SEQ ID NO:21, 1202-1225) and downstream primer (EcoR I) 5'ttatggaattcCGACGCGGCGATTGAGACCTTC (2201-2180, SEQ ID NO:22). C genotype adr serotype HBV g...

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Abstract

The invention relates to peptides derived from HBV L protein for the prevention and treatment of Hepatitis B virus (HBV) infection, screening method for removing antigenicity of these peptides and the deantigenic derivative peptide screened through the method, wherein the HBV surface L protein pre-S1 region contains key amino acid sequence of the virus adhesion cell surface acceptor and antigenic amino acid sequence bonding with the anti-L protein antibody. The polypeptides at the HBV surface L protein pre-S1 region can suppress HBV infection to cells. The invention also provides the screening method for removing the antigenicity of these polypeptides.

Description

technical field [0001] The present invention relates to HBV surface L protein-derived peptides for preventing and treating hepatitis B virus (human hepatitis B virus, HBV) infection, a screening method for removing the antigenicity of these polypeptides, and the deantigenic derived peptides obtained by screening through the method. In particular, the present invention relates to L protein pre-S1 region polypeptides derived from HBV of B genotype adw serotype, C genotype adr serotype and D genotype ayw serotype HBV. Also in particular, the present invention relates to a screening method for removing the antigenicity of the HBV surface L protein-derived peptide and the HBV surface L protein-derived antigenic peptide obtained by the method, which can block HBV infection but does not bind to antibodies. The antigen-removing derivative peptide can cooperate with the anti-HBV surface L protein antibody to inhibit HBV infection. Background technique [0002] 1. Epidemiology of HBV...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K14/01G01N33/68A61K38/16A61P1/16A61P31/20
Inventor 刘宏利汪裕尹颖
Owner SHANGHAI HEP PHARMA
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