HCV NS3 Genotype-1b C33C
Escherichia Coli.
Sterile Filtered clear solution.
Protein is >90% pure as determined by 10% PAGE (coomassie staining).
Description
Recombinant Hepatitis C Virus NS3 Genotype-1b C33C (40-315 aa) produced in E. coli having 226 aa.
Recombinant Hepatitis C Virus NS3 Genotype-1b C33C is fused to a 6xHis tag at C-Terminus and purified by proprietary chromatographic technique.
Product Specs
Hepatitis C Virus (HCV) is a small, enveloped virus with a single-stranded RNA genome. It belongs to the Flaviviridae family and is known for its high replication rate. HCV's RNA polymerase lacks proofreading ability, leading to frequent mutations and contributing to its ability to evade the immune system. The virus is classified into six genotypes (1-6), with variations in their global distribution. Genotype determination is crucial for treatment decisions, as it influences the effectiveness of interferon-based therapy. Genotypes 1 and 4 show lower response rates to this treatment compared to genotypes 2, 3, 5, and 6. HCV NS3 protein of genotype 1b consists of two domains: serine protease and helicase. The C33c fragment of NS3 contains major epitopes targeted by HCV antibodies.
This product consists of a recombinant fragment of the Hepatitis C Virus NS3 protein, specifically from genotype 1b, spanning amino acids 40 to 315 (C33C region). The 226 amino acid protein was produced in E. coli and includes a 6xHis tag fused at the C-terminus. Purification is achieved using a proprietary chromatographic technique.
Sterile Filtered clear solution.
The HCV NS3 Genotype-1b C33C protein is supplied in a solution containing 50mM potassium phosphate buffer.
For short-term storage (2-4 weeks), keep refrigerated at 4°C. For extended storage, freeze at -20°C. Adding a carrier protein like HSA or BSA (0.1%) is recommended for long-term storage. Minimize freeze-thaw cycles to maintain protein integrity.
The purity of this protein is greater than 90%, as assessed by 10% SDS-PAGE followed by Coomassie blue staining.
This product is suitable for use in ELISA and lateral flow assays.
Escherichia Coli.
Product Science Overview
Hepatitis C Virus (HCV) is a significant global health concern, affecting millions of individuals worldwide. The virus is known for causing chronic liver diseases, including cirrhosis and hepatocellular carcinoma. HCV is a single-stranded RNA virus that encodes a large polyprotein, which is processed into structural and nonstructural proteins. Among these, the nonstructural protein 3 (NS3) plays a crucial role in the viral life cycle.
The NS3 protein of HCV is a multifunctional enzyme with serine protease, helicase, and NTPase activities. The N-terminal domain of NS3 contains the serine protease activity, which is essential for the cleavage of the HCV polyprotein into functional units. The C-terminal domain possesses RNA helicase and NTPase activities, which are vital for viral RNA replication .
HCV exhibits significant genetic diversity, with multiple genotypes and subtypes. Genotype-1b is one of the most prevalent and clinically significant genotypes. The C33C region of NS3, spanning amino acids 1192 to 1459, is an immunodominant region recognized by the immune system of HCV-infected individuals . This region is often targeted in diagnostic assays and vaccine development due to its immunogenic properties.
Recombinant NS3 C33C is a laboratory-produced version of the NS3 protein, specifically from the genotype-1b strain of HCV. This recombinant protein includes the full-length C33C immunodominant region. It is used in various research and diagnostic applications, including enzyme-linked immunosorbent assays (ELISA) and Western blotting, to detect HCV antibodies in patient sera .
The production of recombinant NS3 C33C involves several steps, including gene cloning, expression, and purification. The gene encoding the NS3 C33C region is cloned into an expression vector, which is then introduced into a host cell, such as Escherichia coli or mammalian cells. The host cells express the recombinant protein, which is subsequently purified using techniques like affinity chromatography .
The synthetic route for producing recombinant NS3 C33C typically involves the following steps:
- Gene Cloning: The gene encoding the NS3 C33C region is synthesized and inserted into an expression vector.
- Transformation: The expression vector is introduced into a host cell, such as E. coli, through a process called transformation.
- Expression: The host cells are cultured under specific conditions to induce the expression of the recombinant protein.
- Purification: The recombinant protein is purified from the host cell lysate using affinity chromatography, which exploits the specific binding properties of the protein .
The reaction conditions for each step are optimized to ensure high yield and purity of the recombinant protein. For example, the expression conditions may include specific temperature, pH, and nutrient concentrations to maximize protein production .
Recombinant NS3 C33C is widely used in research to study the structure and function of the NS3 protein, as well as in the development of diagnostic assays and vaccines. Its immunogenic properties make it a valuable tool for detecting HCV infections and understanding the immune response to the virus .
