Description
General description
Genome polyprotein (UniProt: P27958) is encoded by the POLY gene (Gene ID: 951475) in Hepatitis C virus. HCV is a small (~55-65 nm), enveloped, positive-sense single-stranded RNA virus that is a causative factor for hepatitis C and hepatocellular carcinoma. HCV expresses several proteins that promote viral replication. Genome polyprotein of HCV can be cleaved into 11 different chains. The mature core protein packages viral RNA to form a viral nucleocapsid and promotes virion budding. It also prevents the establishment of cellular antiviral state by blocking the interferon- /β and interferon- signaling pathways and by inducing STAT1 degradation. Envelope proteins E1 and E2 that form a heterodimer are heavily glycosylated and are involved in virus attachment to the host cell, virion internalization through clathrin-dependent endocytosis and fusion with host membrane. In HCV, Viroprotein P7, a heptameric ion channel protein, plays an essential role in the assembly, envelopment, and secretion of viral particles. It also contains NS2, a cysteine protease and NS3, a serine protease. NS2 is required for the proteolytic auto-cleavage between NS2 and NS3. NS3 binds a single ATP and catalyzes the unzipping of a single base pair of dsRNA. Non-structural protein 4A (NS4A), a peptide cofactor, forms a non-covalent complex with the N-terminal of NS3 and this complex prevents phosphorylation of host IRF3, which also prevents the establishment of dsRNA induced antiviral state. NS4B induces a specific membrane alteration that serves as a scaffold for the virus replication complex. Non-structural protein 5A (NS5A), a multi-functional phosphoprotein, is an RNA-binding peripheral membrane protein that is reported to antagonize numerous cellular pathways, including the antiviral interferon-a response. NS5A exists as hypo- and hyper-phosphorylated forms and the dynamic transition between these two states is involved in the functions of NS5A. Hyperphosphorylation occurs primarily at its six serine residues within the low complexity sequence of NS5A. Phosphorylated NS5A is shown to be essential for viral replication and assembly. Clone AR4A recognizes a discontinuous epitope on the E1E2 complex. It is shown to bind only to the folded E1E2 complex and does not block binding of E1E2 to CD81. It displays exceptionally broad neutralizing activity toward diverse HCV genotypes and protects against heterologous HCV challenge in small animal models. (Ref.: 1090-1094; Giang, E., et al. (2012). Proc. Natl. Acad. Sci. USA. 109(16); 6205-6210; Law, M., et al. (2008). Nat. Med. 14(1); 25-27).
Immunogen
From Fab phage display library generated from the bone marrow of a patient with chronic HCV infection.
Application
Quality Control Testing
Evaluated by ELISA in lysate from HEK293T cells transfected with H77E1E2.
ELISA Analysis (ELISA): Envelope glycoprotein E1E2 was detected by ELISA in various dilutions of lysates from HEK293T cells transfected with H77E1E2, but not in lysate from untransfected cells.
Tested Applications
ELISA Analysis: A representative lot detected HCV E1E2 in ELISA applications (Giang, E., et al. (2012). Proc Natl Acad Sci USA. 109(16):6205-10).
Immunoprecipitation Analysis: A representative lot immunoprecipitated HCV E1E2 in Immunoprecipitation applications (Giang, E., et al. (2012). Proc Natl Acad Sci USA. 109(16):6205-10).
Neutralizing: A representative lot of this antibody neutralized several HCV isolates (Giang, E., et al. (2012). Proc. Natl. Acad. Sci. USA. 109(16):6205-10).
Note: Actual optimal working dilutions must be determined by end user as specimens, and experimental conditions may vary with the end user.
Biochem/physiol Actions
Clone AR4A is a human recombinant antibody that detects envelope glycoproteins E1FE2 of Hepatitis C virus. It recognizes a discontinuous epitope from the N-terminal half. It binds to the folded E1E2 complex.
Physical form
Purified human recombinant monoclonal antibody IgG1 in PBS without azide
Preparation Note
1.0 mg/mL. Please refer to guidance on suggested starting dilutions and/or titers per application and sample type.
Store at -10°C to -25°C. Handling Recommendations: Upon receipt and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance.
Other Notes
Concentration: Please refer to the Certificate of Analysis for the lot-specific concentration.
Disclaimer
Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
