Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Antiviral protein and uses thereof

A protein and anti-virus technology, applied in the protein field, can solve problems such as inability to encode viral proteins, G→A high-frequency mutation, truncated non-functional products, etc., to reduce immunogenicity, improve membrane penetration efficiency, facilitate purification or Detection effect

Inactive Publication Date: 2012-05-23
SHANTOU UNIV MEDICAL COLLEGE
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] or Truncated nonfunctional product due to premature formation of the stop codon
[0013] In addition, APOBEC has a large molecular weight and strong immunogenicity, and direct application to the human body may induce high antibody levels and reduce its antiviral effect

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Antiviral protein and uses thereof
  • Antiviral protein and uses thereof
  • Antiviral protein and uses thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1 Preparation of antiviral protein

[0039] The amino acid sequence of the antiviral protein of the present embodiment is:

[0040] MetGlySerSer HisHisHisHisHisHis SerSerGlyLeuValProArgGlySerHisMetAlaSerMetThrGlyGlyGlnGlnMetGlyArgGlySer lyGlyGlyGlySerValAspLeuGlu HisHisHisHisHisHis

[0041]This antiviral protein contains a membrane-penetrating peptide domain (GlyArgLysLysArgArgGlnArgArgArgProProGln) and three cytosine deaminase domains derived from the APOBEC family (these three cytosine deaminase domains are sequentially derived from the cytosine at the N-terminal of the hAPOBEC3G molecule 脱氨酶结构域HisProGluMetArgPhePheHisTrpPheSerLysTrpArgLysLeuHisArgAspGlnGluTyrGluValThrTrpTyrIleSerTrpSerProCysThrLysCys、来自hAPOBEC3G分子C端的胞嘧啶脱氨酶结构域HisAlaGluLeuCysPheLeuAspValIleProPheTrpLysLeuAspLeuAspGlnAspTyrArgValThrCysPheThrSerTrpSerProCysPheSerCys、来自hAPOBEC3F分子C端的胞嘧啶脱氨酶结构域HisAlaGluArgCysPheLeuSerTrpPheCysAspAspIleLeuSerProAsnThrAsnTyrGluValThrTrpTyrThrSerTrpSerProCysProG...

Embodiment 2

[0103] Example 2 Western blot detection of the antiviral protein prepared in Example 1

[0104] The purified antiviral protein was added to the HepG2.2.15 cells growing into a monolayer at a final concentration of 10 μg / ml (that is, the HepG2.2.15 cells growing into a monolayer finally contained 10 μg of purified antiviral protein), As the experimental group; add PBS to HepG2.2.15 cells in the control group. After 8 hours, the cells were lysed to extract the total protein, and Western blot was performed with an anti-His tag mouse monoclonal antibody, and detected by chemiluminescence. like image 3 ( image 3 Among them, M represents protein molecular weight standard; Swimming lane 1. Western blot result of HepG2.2.15 cells in the control group; Swimming lane 2. As shown in the Western blot result of HepG2.2.15 cells in the experimental group adding antiviral protein), the cells in the experimental group appeared at 22KD There was a distinct band A, consistent with the mole...

Embodiment 3

[0105] Example 3 Analysis of intracellular localization of the antiviral protein prepared in Example 1

[0106]Add the purified antiviral protein to the HepG2.2.15 cell slides growing into a monolayer at a final concentration of 5 μg / ml (that is, each ml of HepG2.2.15 cell slides growing into a monolayer contains 5 μg of purified antiviral protein) as the experimental group; add PBS to the HepG2.2.15 cell slides of the control group. After 8 hours, the cell slides were fixed with 4% paraformaldehyde at room temperature for 10 min; then 0.2% Triton-X100 (Triton-X100) was added, and fixed at room temperature for 5 min; then, anti-His tag The mouse monoclonal antibody was used as the primary antibody, and the Cy3-labeled goat anti-mouse antibody was used as the secondary antibody. Observed under a fluorescent microscope, such as Figure 4 as shown ( Figure 4 The middle red part shows the antiviral protein that enters the cytoplasm), and it was found that the antiviral protein ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to anti-viral protein. The anti-viral protein is characterized in that the anti-viral protein is fusion protein and comprises a transmembrane peptide structural domain and a tandem body which is formed through at least two cytosine deaminase structural domains originated from an APOBEC family, a transmembrane peptide structural domain and various cytosine deaminase structural domains. The transmembrane peptide structural domain in the anti-viral protein provided by the invention ensures the anti-viral protein to have the transmembrane capability and to freely shuttle among cells and enters the cells in a non-receptor and non-energy dependent way; the cytosine deaminase structural domains originated from the APOBEC family ensures that the anti-viral protein can effectively inhibit the reproduction of human immunodeficiency virus and / or hepatitis B virus after entering cells; because the non-essential sequences in the APOBEC family protein molecules are removed, not only the antiviral activity of the APOBEC family protein molecules is preserved, but also the immunogenicity of the anti-viral protein is reduced, and the transmembrane efficiency of the anti-viralprotein is enhanced.

Description

technical field [0001] The present invention relates to protein, in particular, relates to an antiviral protein for treating human immunodeficiency virus (HIV, human immunodeficiency virus) and / or hepatitis B virus (HBV, hepatitis B virus) infection, and this antiviral protein Applications of viral proteins. Background technique [0002] APOBEC family (apolipoprotein B mRNA editing enzyme-catalytic polypeptide family, apolipoprotein B mRNA editing enzyme catalytic polypeptide family) is a newly discovered protein molecule with cytosine deaminase activity in recent years, which can convert cytosine deamination It is uracil (C→U), and its family members include: hAPOBEC1 (human APOBEC1), hAPOBEC2 (human APOBEC2), hAPOBEC3A (human APOBEC3A), hAPOBEC3B (human APOBEC3B), hAPOBEC3C (human APOBEC3C), hAPOBEC3D (i.e. human APOBEC3D), hAPOBEC3E (i.e. human APOBEC3E), hAPOBEC3F (i.e. human APOBEC3F), hAPOBEC3G (i.e. human APOBEC3G), hAPOBEC3H (i.e. human APOBEC3H), hAPOBEC4 (i.e. hum...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C07K19/00A61K38/17A61P31/18A61P31/20
Inventor 李卫中李康生王革非辛岗苏芸陈小璇陈幼莹张衡张丹桂曾俊
Owner SHANTOU UNIV MEDICAL COLLEGE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com