The authors have paid a fee to allow immediate free access to this article.
Affinity maturation by light-chain replacement. (A) Three VL chains from the designated neutralizing HC33 HMAbs that were paired to three nonneutralizing VH chains and the designations of nine VHVL derivative clones. (B) Neutralization of 2a JFH1 HCVcc by affinity-matured HC33 HMAb clones at 20 μg/ml. The assay was performed as described for Fig. 1A. HC33.1 and R04 were used as positive and negative controls, respectively. Data are shown as percent neutralization (mean ± SD of results from three experiments). (C) Affinity-matured HC33.6 HMAb clones block E2 binding to CD81. Genotype 1a H77C E1E2 lysate containing 1 μg/ml E2 was incubated with each test HMAb at 5 and 15 μg/ml, and the antibody-antigen complex was then added onto CD81-LEL-precoated microtiter wells. The assay was performed as described for Fig. 1D. HC-11 and R04 were used as positive and negative controls, respectively. The data are shown as percent inhibition (mean ± SD of results from two experiments performed in triplicate). (D) Epitope mapping of the affinity-matured antibodies in the E2 region of aa 412 to 423. The assay was performed as described for Fig. 2D.