Recombinant Human Anti-HCV E2 Antibody (HC33.1) (CAT#: PABL-119)

Figure 2 (A) Epitope alignment. Epitopes of three HMAbs: HC33.1; HC33.4 and HC33.8 are compared with murine MAb AP33. Recombinant E1E2 mutant proteins were expressed in 293T cells and cell lysates were analyzed by ELISA. Individual protein expression was normalized by binding of CBH-17, an anti-HCV E2 HMAb to a linear epitope. Red indicates 0–20%, orange 21–40%, brown 41–60%, white 61–100% and green >100% binding, when the residue was replaced by alanine, relative to binding to wt.

Keck, Z. Y., Angus, A. G., Wang, W., Lau, P., Wang, Y., Gatherer, D.,... & Foung, S. K. (2014). Non-random escape pathways from a broadly neutralizing human monoclonal antibody map to a highly conserved region on the hepatitis C virus E2 glycoprotein encompassing amino acids 412–423.PLoS pathogens, 10(8), e1004297.

Figure 3 Dose-dependent neutralization of (B) wt JFH-1 HCVcc, (C) HCVcc variant bearing N417S mutation and (D) bearing N417T mutation were performed by SEAP reporter assay. (B, C, D) Either wt HCVcc or variant HCVcc was incubated with HC33.1 or AP33, at concentrations ranging from 0.1 to 50 µg/ml, prior to infecting the Huh7J-20 cells. Virus infectivity levels were determined by measurement of the SEAP activity released into the medium.

Keck, Z. Y., Angus, A. G., Wang, W., Lau, P., Wang, Y., Gatherer, D.,... & Foung, S. K. (2014). Non-random escape pathways from a broadly neutralizing human monoclonal antibody map to a highly conserved region on the hepatitis C virus E2 glycoprotein encompassing amino acids 412–423.PLoS pathogens, 10(8), e1004297.

Figure 4 (A) Dual antibody immunofluorescence staining of Huh7.5 cells infected with JFH1 2a HCVcc during multiple passages in increasing concentration of HC33.1. IFA is shown for P0, P18, P27 and P31. HCV E2 glycoprotein was stained with HC33.1 under which viral escape variants were selected (green, upper set of panels), or with CBH-5, a neutralizing domain B HMAb that does not share the same epitope with HC33.1 on E2 (green, lower set of panels). Total virus-infected cells were stained with anti-NS3 antibody labeled with Alexa-594 (red). The cells were counterstained with Hoechst nuclear stain H33342 (blue). The captured images were superimposed (merge).

Keck, Z. Y., Angus, A. G., Wang, W., Lau, P., Wang, Y., Gatherer, D.,... & Foung, S. K. (2014). Non-random escape pathways from a broadly neutralizing human monoclonal antibody map to a highly conserved region on the hepatitis C virus E2 glycoprotein encompassing amino acids 412–423.PLoS pathogens, 10(8), e1004297.

Figure 5 (A) HC33.1 dose-dependent neutralization against viral pool in culture supernatants collected from P0, P18, P27 and P34 with their respective dominant variants bearing the following mutations: N417S, N417T/N434D/K610R, N417T/S419N/N434D/K610R and S395/L413I/N417T was performed by FFU-reduction assay.

Keck, Z. Y., Angus, A. G., Wang, W., Lau, P., Wang, Y., Gatherer, D.,... & Foung, S. K. (2014). Non-random escape pathways from a broadly neutralizing human monoclonal antibody map to a highly conserved region on the hepatitis C virus E2 glycoprotein encompassing amino acids 412–423.PLoS pathogens, 10(8), e1004297.

Figure 6 Dose-dependent neutralization against recombinant HCVcc variants bearing specific mutations, as identified in each phase of viral escape selection was performed by SEAP reporter assay. The IC50 value against each variant is tabulated in the legend.

Keck, Z. Y., Angus, A. G., Wang, W., Lau, P., Wang, Y., Gatherer, D.,... & Foung, S. K. (2014). Non-random escape pathways from a broadly neutralizing human monoclonal antibody map to a highly conserved region on the hepatitis C virus E2 glycoprotein encompassing amino acids 412–423.PLoS pathogens, 10(8), e1004297.