FIG. 3.

Nine HCV HMAbs fail to recognize denatured HCV E2. Proteins derived from HeLa cells infected with vaccinia virus Q1b and VWA or VWA alone (gray bars) were either left untreated (white bars) or denatured by incubation with 0.5% sodium dodecyl sulfate and 5 mM dithiothreitol for 15 min at 56°C (black bars). After treatment, proteins were diluted 1:5 in BLOTTO and captured onto wells coated with 500 ng of GNA. Wells were washed and blocked, and bound protein was incubated at 5 μg/ml with HCV HMAbs (x axis) and control HMAb (R04). Bound antibody was detected as described in Materials and Methods. Values for native and denatured HCV 1b are the mean signals obtained from replicate wells. Signals from single wells of native and denatured proteins derived from VWA-infected HeLa cells were indistinguishable and also averaged. Error bars indicate 1 standard deviation from the mean.