HCV NS3

Hepatitis C Virus NS3 (1450-1643 a.a.) Recombinant
Cat. No.
BT13292
Source
Synonyms
Appearance
Purity
HCV NS3 protein is >95% pure as determined by 10% 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 22kDa recombinant protein contains the HCV NS3 genotype 1b immunodominant regions, amino acids 1450-1643. The protein is fused with b-galactosidase (114 kDa) at N-terminus, pI 5.43.

Product Specs

Introduction
HCV is a small, 50nm, enveloped, single-stranded, positive-sense RNA virus in the family Flaviviridae. It has a high replication rate, producing approximately one trillion particles daily in an infected individual. The HCV RNA polymerase lacks proofreading ability, resulting in a high mutation rate that may help the virus evade the host's immune response. Hepatitis C virus is classified into six genotypes (1-6), each with several subtypes. The prevalence and distribution of HCV genotypes vary globally. Genotype is clinically significant in determining the potential response to interferon-based therapy and its required duration. Genotypes 1 and 4 are less responsive to interferon-based treatment than genotypes 2, 3, 5, and 6.
Description
The E. coli-derived recombinant HCV NS3 protein (22 kDa) encompasses the genotype 1b immunodominant regions, amino acids 1450-1643. This protein is N-terminally fused with b-galactosidase (114 kDa) and has a pI of 5.43.
Purity
HCV NS3 protein purity is greater than 95% as determined by 10% SDS-PAGE (Coomassie staining).
Formulation
20mM Tris-HCl (pH 8.0), 8M urea, and 10mM β-mercaptoethanol.
Stability
HCV NS3 is stable at 4°C for one week but should be stored below -18°C. Avoid repeated freeze-thaw cycles.
Applications
HCV NS3 antigen is suitable for use in ELISA and Western blots. It serves as an excellent antigen for HCV detection with minimal specificity issues.
Purification Method
HCV NS3 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, infecting millions of people worldwide. It is a positive, single-stranded RNA virus belonging to the Flaviviridae family. The HCV genome encodes a single polyprotein of approximately 3,000 amino acids, which is processed by host and viral proteases into structural and non-structural proteins. Among these, the non-structural protein 3 (NS3) is particularly noteworthy due to its multifunctional enzymatic activities and its role in viral replication and pathogenesis.

Structure and Function

The NS3 protein of HCV is a multifunctional enzyme with three distinct activities: serine protease, NTPase, and RNA helicase. The serine protease activity of NS3 is responsible for the proteolytic processing of the HCV polyprotein, which is essential for the maturation of other non-structural proteins, including NS4A, NS4B, NS5A, and NS5B . The NTPase and RNA helicase activities are crucial for viral RNA replication.

The NS3 protein spans amino acids 1027 to 1657 of the HCV polyprotein. The recombinant fragment of NS3, encompassing amino acids 1450 to 1643, retains the essential enzymatic activities and is often used in research to study the protein’s function and to develop antiviral drugs .

Preparation Methods

The recombinant NS3 protein (1450-1643 a.a.) is typically expressed in Escherichia coli (E. coli) systems. The gene encoding the NS3 fragment is cloned into an expression vector, which is then introduced into E. coli cells. The bacteria are cultured, and the expression of the recombinant protein is induced. Following induction, the cells are lysed, and the NS3 protein is purified using chromatographic techniques to achieve high purity levels, often exceeding 95% .

Applications and Significance

The recombinant NS3 protein is widely used in various research applications, including:

  • Enzyme Activity Assays: To study the protease, NTPase, and helicase activities of NS3.
  • Drug Screening: To identify and develop inhibitors targeting the NS3 protease, which is a key therapeutic target for HCV treatment.
  • Immunological Studies: To investigate the immune response to HCV infection and to develop diagnostic assays for detecting HCV antibodies .

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