FIG. 6.

H-111 epitope mapping. (A) HEK293 cells transfected with HCV E1 constructs comprising the E1 gene deletions shown were analyzed by IFA and confirmed with complete agreement by Western blotting (data not shown). The colored bar on the E1 deletion of amino acids 192 to 305 shows the specific epitope location. (B) Competition analysis of the binding of H-111 to HEK293 cells expressing E1 with overlapping synthetic peptides by ELISA. Wells were incubated with 10 μg of H-111/ml in the presence of increasing amounts (x axis) of blocking peptides. ○, control HTLV peptide; ▴, E1 peptide YEVRNVS; ▪, E1 peptide YEVRNVSGVYHVTN. The y axis shows the mean optical density values for triplicate wells. Error bars represent one standard deviation from the mean. (C) Alanine scanning of H-111 epitope by ELISA. The amino acid sequence of the H-111 epitope is shown in the top row. The localization of alanine substitution is indicated in each subsequent row. The amino acid sequence of the H-111 epitope on the E1 glycoprotein is indicated on top in single-letter code. The x axis values are relative optical density values for binding normalized to that for the nonsubstituted sequence (top row), which was set at 100%. The value for binding to each substituted protein is the mean for three separate studies and is shown with a range bar.