HCV NS5 Genotype-1b

Hepatitis C Virus NS5 Genotype-1b Recombinant
Cat. No.
BT16299
Source
Escherichia Coli.
Synonyms
Appearance
Sterile Filtered clear solution.
Purity
HCV NS5 Genotype-1b protein is >95% pure as determined by SDS- PAGE (coomassie staining).
Usage
THE BioTek's products are furnished for LABORATORY RESEARCH USE ONLY. The product may not be used as drugs, agricultural or pesticidal products, food additives or household chemicals.
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Description

The E.coli derived recombinant protein contains the HCV NS5 Genotype1b immunodominant regions, amino acids 2212-2313. The protein is fused to a GST tag at N-terminus.

Product Specs

Introduction
Hepatitis C virus (HCV) is a small (50nm), enveloped, single-stranded RNA virus with a positive sense genome. It belongs to the family Flaviviridae. HCV replicates rapidly, producing approximately one trillion particles daily in an infected person. Its RNA polymerase lacks proofreading ability, leading to a high mutation rate. This characteristic contributes to the virus's ability to evade the host's immune system. HCV is classified into six genotypes (1-6), each with multiple subtypes. The prevalence and distribution of these genotypes vary globally. Genotyping is clinically significant as it guides treatment decisions, particularly for interferon-based therapy. Genotypes 1 and 4 respond less favorably to interferon compared to genotypes 2, 3, 5, and 6, necessitating adjustments in treatment duration and strategies.
Description
This recombinant protein is derived from E. coli and encompasses the immunodominant regions of the HCV NS5 protein from genotype 1b, specifically amino acids 2212-2313. A GST tag is fused to the N-terminus of the protein.
Purity
The purity of the HCV NS5 Genotype-1b protein exceeds 95%, as determined by SDS-PAGE analysis with Coomassie blue staining.
Physical Appearance
The product is a clear, sterile-filtered solution.
Formulation
The protein is supplied in a buffer containing 1.5M urea, 25mM Tris-HCl (pH 8.0), 0.2% Triton-X, and 50% glycerol.
Stability
For optimal stability, HCV NS5 Genotype-1b should be stored at -18°C. Although it can remain stable at 4°C for up to one week, repeated freeze-thaw cycles should be avoided.
Applications
The HCV NS5 Genotype-1b antigen is suitable for use in a variety of research applications, including ELISA and Western blots. Its high specificity makes it an excellent choice for the detection of HCV.
Source
Escherichia Coli.
Purification Method
HCV NS5 Genotype-1b protein was purified by proprietary chromatographic technique.
Specificity
Immunoreactive with sera of HCV-infected individuals.

Product Science Overview

Introduction

Hepatitis C virus (HCV) is a significant global health concern, affecting over 150 million people worldwide. It is a major cause of chronic liver diseases, including cirrhosis and hepatocellular carcinoma (HCC). HCV is classified into seven genotypes, with numerous subtypes that respond differently to antiviral therapies. Among these, genotype 1, particularly subtype 1b, is the most prevalent worldwide .

Hepatitis C Virus Structure and Genotypes

HCV is an RNA virus belonging to the Flaviviridae family. Its genome encodes a single polyprotein that is processed into structural and non-structural proteins. The non-structural protein 5 (NS5) is a crucial component of the HCV replication complex. It consists of two subunits: NS5A and NS5B. NS5A is a multifunctional phosphoprotein involved in viral replication and assembly, while NS5B is an RNA-dependent RNA polymerase essential for viral RNA synthesis .

Recombinant HCV Systems

The development of recombinant HCV infectious cell culture systems has been a significant advancement in the study of HCV. These systems allow for the replication and study of different HCV variants in vitro. The high replication capacity of the JFH1 strain (genotype 2a) has been particularly useful in developing these systems. However, efficient infectious JFH1-based cell cultures of subtype 1b have been limited .

Development of NS5 Genotype-1b Recombinant

Researchers have developed efficient infectious JFH1-based cultures with genotype 1b core-NS5A sequences of strains DH1, Con1, and J4. This was achieved by using previously identified HCV cell culture adaptive substitutions A1226G, R1496L, and Q1773H. These viruses spread efficiently in Huh7.5 cells by acquiring additional adaptive substitutions, and the final recombinants yielded peak supernatant infectivity titers of 4 to 5 log 10 focus-forming units (FFU)/ml .

Applications and Significance

The development of these recombinant HCV systems is crucial for several reasons:

  1. Efficacy Testing: They allow for the efficacy testing of various classes of antivirals, including NS3/4A protease and NS5A inhibitors. These inhibitors have shown significant efficacy against genotype 1 viruses .
  2. Resistance Studies: Recombinant systems enable the study of resistance mechanisms to antiviral therapies, which is essential for developing more effective treatments .
  3. Virological Studies: These systems provide valuable tools for other virological studies requiring genotype 1b infectious viruses .

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