United Kingdom (UK)
Recombinant monoclonal antibody to Glycoprotein E2. Manufactured using AbAb’s Recombinant Platform with variable regions (i.e. specificity) from the hybridoma AP33.
UniProt Accession Number of Target Protein: P27958
Alternative Name(s) of Target: NS1; gp68; gp70; HCV glycoprotein E2; Genome polyprotein; Envelope glycoprotein E2; E2 glycoprotein
Immunogen: The original antibody was generated by immunizing a mouse with HCV envelope glycoprotein E2.
Specificity: This antibody recognizes a linear epitope comprising residues 412 to 423 of HCV envelope glycoprotein E2. The hepatitis C virus is the cause of hepatitis C and some cancers such as liver cancer (hepatocellular carcinoma, abbreviated HCC) and lymphomas in humans.
Application Notes: The binding of AP33 inhibits the interaction between CD81 and a range of presentations of E2, including soluble E2, E1E2, and virus-like particles. To asses and the reactivity and neutralizing capacity of antisera and monoclonal antibodies, proteins from HEK293T cells cotransfected with a plasmid containing HCV genotype 1a strain H77-derived full-length E1E2 were immunoprecipitated using the mouse version of this antibody. Further on the same HEK293T cells an ELISA was performed using the mouse version of this antibody. Finally a neutralization assay was performed by incubating huh-7 cells with HCVpp in the presence of the mouse version of this antibody and then measuring the fluorescence using flow cytometry (Owsianka et al, 2005; pmid:16103160). To measure the relative binding affinity of the mouse version of this antibody to HCV E1E2 glycoproteins, a cell lysate–based ELISA was performed. Further HCV pseudo particles (HCVpp) and cell-culture–derived HCV (HCVcc) neutralization assays were performed (Desombere et al, 2016; pmid:26710081). To determine the reactivity of selected phage with the selecting antibody, a phage capture enzyme immunoassay was performed. For this assay the mouse version of this antibody was used. Further to detect MAb binding to E2 glycoprotein, an ELISA was performed on transfected HEK293FT cells using the mouse version of this antibody (Tarr et al, 2006; pmid:16496330). Binding of the mouse version of this antibody was assessed in the presence of the peptide-reactive serum, and HCV-positive, peptide-unreactive serum and normal human serum. Further to determine the neutralization ability of this antibody a neutralization assay was performed on huh-7 cells and HEK293T cells infected with HCVpps (Tarr et al, 2007; pmid:17947521).
Antibody first published in:
Owsianka et al. Monoclonal antibody AP33 defines a broadly neutralizing epitope on the hepatitis C virus E2 envelope glycoprotein. J Virol. 2005 Sep;79(17):11095-104. PMID:16103160
Note on publication:
The generation of retrovirus-based pseudo particles (HCVpp) incorporating a panel of full-length E1E2 clones representative of the major genotypes 1 through 6, and their application to assess the reactivity and neutralizing capability of antisera and monoclonal antibodies raised against portions of the HCV E2 envelope protein is described.