Recombinant antibody Fab against the hypervariable region 1 of hepatitis C virus blocks the virus adsorption to susceptible cells in vitro

Antibodies against hypervariable region 1 (HVR1) of hepatitis C virus (HCV) are putatively considered to be neutralizing. We previously found that monoclonal antibodies (mAbs) (30F1 and 30F3) against the HVR1 of HCV neutralize HCV in vitro. To develop potentially therapeutic molecules against HCV, we cloned cDNAs of antibody Fab fragments from the mouse hybridoma cells secreting these two mAbs. Fab fragments produced in Escherichia coli were purified by a single step of nickel-chelate affinity chromatography via a hexa-histidine tag. The specificity of the Fabs was confirmed by competition ELISA, BIAcore analysis, and N-terminal amino acid sequencing. The binding constant for the interaction with HVR1 was 1.39 nM for Fab 30F1 and 3.96 nM for Fab 30F3. The HCV capture assay and inhibition of HCV adsorption test demonstrated that both Fabs had neutralizing activity. The data may be useful for designing immunological therapy of HCV.