Hepatitis C

Accession #/Synonym:  core protein p21, capsid protein C, envelope gpE1, NS1, gp68, NS5a, NS3P, p23,.

Worldwide 170 million people are affected by HCV, a major cause of chronic Hepatitis and in severe sequel may lead of hepatic cirrhosis, hepatocellular carcinoma and death.  Hepatitis C Virus (HCV) is a small RNA enveloped, positive sense genome that encodes for the replication and assembly of core protein (1). HCV regulates hosts non coding RNAs to promote viral replication.  HCV is a member of Flaviviridae family, genus Hepacivirus whose most common method of transmission is through blood or blood products.  HCV infection is characterized by an increase in free radical formation manifested by increased hepatic and serum levels of products of lipid peroxidation. Hepatitis C has been implicated in chronic kidney disease (CKD), a common disease in the United States, affecting nearly 11% of the adult population. HBV and HIV are highly coinfected with HCV in endemic areas where it increases the risk of liver failure; hepatocellular carcinoma and therapeutic failure. Antibody-mediated inhibition of cellular HCV-Like Particles binding represents a convenient system for the functional characterization of human anti-HCV antibodies, allowing the mapping of envelope neutralization epitopes targeted by naturally occurring antiviral antibodies.

Upon infection, HCV directly alters host cell lipid metabolism through nuclear transcription factor.  Many nutrients like β-carotene, vitamin D2 & linoleic acid serves to decrease viral replication. Polyunsaturated fatty acid with high concentration of omega 3 fatty acids suppresses mRNA expression of metabolic enzymes leading to decrease hepatic lipogenesis and triglyceride synthesis.  Many antivirus drugs like ribavirin and interferon’s are in use for  treatment but their action varies according to viral genome, host factor, dosage schedule and others. Viral entry is a major target of neutralizing antibodies; a first-line host defense inhibiting viral spread. Rapid induction of cross-neutralizing antibodies in the very early phase of infection has been suggested to control of HCV infection.

HepC antibodies were made against polyprotein that codes for C, E1, E2, p7, NS2, NS3, NS4a, NS4b, NS5a, and NS5b. antigenic peptide was selected form the region spanning aa 2198-2216.  The HepC polyprotein antibodies were affinity purified on immobilized antigen based affinity chromatography. Hep C affinity purified antibodies are stabilized in antibody stabilization buffer for long-term storage.  FabGenix will conjugate HepC antibodies to fluorophores and secondary enzymes (horseradish peroxidase and alkaline phosphatase) at nominal cost.  FabGennix will also provide limited quantities of antigenic blocking peptide (please inquire availability before ordering). 

Immunogen:        Synthetic peptide corresponding to amino acid 2198-2217 of hepatitis C polyprotein corresponding to amino acids (CGPSPPSLASSSASQLSAPSLK).  The peptide was post synthetically modified and conjugated to a carrier protein using hetero-bifunctional cross linker for preparation of antigen to immunize rabbits. 

Concentration:     HepC-101AP concentration was 0.5-0.65mg/ml in antibody stabilization buffer.

Applications:        ELISA/dot blot:  Antibody dilution 1:20,000-1:50,000;.  Western blot:  Antibody dilution 1:500-750 in diluObuffer for use with PC-HepC sample (20ul/lane).  Other applications (IHC, confocal and Immunoprecipitation for these antibodies is not established.  Use of this antibody in applications not listed here does not necessarily exclude its use in such procedures. Investigators employing this antibody in applications not listed here may request a complimentary sample. HepC-101AP labels several diffuse band between 225Kda-25kDa in PC-HepC sample. 

Protocols:         Standard protocol for various applications (WB; IMM and IHC) of this antibody can be requested by calling Technical service line, basic information on this antibody is provided with the product specification sheet, however, FabGennix Inc. recommends investigators to optimize conditions for use of this antibody in their assay. .  

Form/Storage:  The antiserum is supplied in antibody stabilization buffer with preservatives.  For long-term storage of antibody, store at -20^oC FabGennix Inc. does not recommend storage of very dilute antibody solutions unless they are prepared in specially formulated multi-use antibody dilution buffer (Cat # DiluOBuffer).  Working solutions of antibodies in DiluOBuffer should be filtered through 0.45ƒÝ filter after every use for long-term storage.

 

For research use only, not for diagnostic or therapeutic use.