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Hepatitis B virus surface L protein related peptide

A hepatitis B, sequence technology, applied in the direction of viral peptides, antiviral agents, peptide sources, etc.

Active Publication Date: 2014-08-27
SHANGHAI HEP PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 2. 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

[0085] 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

[0086] 1. Preparation of peptides derived from surface L protein of HBV serotype B genotype adw serotype and C genotype adr serotype

[0087] 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. Ion-free water dissolves peptide cleavage products, Explorer...

Embodiment 2

[0098] Example Two: Inhibition of HBV Hepatocyte Infection by Peptides Derived from Surface L Protein of D Genotype Ayw Serotype HBV Virus

[0099] 1. The synthesis of Myr2-47 (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.

[0100] 2. Cultivation of D genotype ayw serotype HBV virus

[0101] 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 bovine serum, and frozen at -80°C.

[0102] 3. Detection of HBsAg in hepatocyte culture supernatant after HBV infection (same as Example 1)

[0103] 4. Inhibition of HBV hepatocyte infection by HBV surface L protein...

Embodiment 3

[0105] Example 3: Screening of HBV surface L protein derivatized peptides

[0106] 1. Determination of HBV virus genotype and serotype of B genotype adw serotype and C genotype adr serotype, virus culture, and culture of primary human hepatocytes (same as Example 1).

[0107] 2. The construction of a recombinant plasmid carrying a complete HBV genome and having the complete replication ability of HBV.

[0108] Add 310μl proteinase K lysate to 50μl 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) Genotype C adr serot...

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Abstract

The invention relates to hepatitis B virus (HBV) surface L protein derived peptide which is used for preventing and curing HBV infection, a screening method which is used for removing polypeptide antigenicity and antigenicity-removing derived peptide which is screened and obtained through the screening method. An HBV surface L protein pre-S1 area has a key amino acid sequence of virus adherent cell surface receptors and an antigenicity amino acid sequence which is combined with antibody against L protein. Polypeptide in the HBV surface L protein pre-S1 area can inhibit the infection of the HBV on cells. The invention also provides the screening method which is used for removing polypeptide antigenicity. The antigenicity-removing derived peptide is screened, can inhibit HBV infection and does not combine with the antibody. Therefore, the mutual interference between natural L protein derived peptide and the antibody against the L protein during HBV infection inhibition is avoided, and both the antigenicity-removing derived peptide and the antibody against the HBV surface L protein can inhibit the HBV infection.

Description

[0001] The patent application of the present invention is a divisional application of the invention patent application submitted by the applicant on August 12, 2005, with the application number 200510028721.4 and the invention title "Hepatitis B virus surface L protein-related peptide". technical field [0002] The present invention relates to HBV surface L protein-derived peptides used 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 de-antigenic 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 protei...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K14/02A61K39/29A61P1/16A61P31/20
Inventor 刘宏利汪裕尹颖
Owner SHANGHAI HEP PHARMA
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