Anti-HIV-1 envelope mAbs produced from single IgG+ memory B cells selected using JRFL expressing virus-like particles.

1A standard ELISA was used to determine the binding activity of mAbs to antigens. Monoclonal Abs were tested at a concentration of 10 µg/ml against gp120 and gp41 coated onto ELISA plate at 1 µg/ml; cardiolipin at a concentration of 45 µg/ml in ethanol was coated by evaporation at 4°C overnight. The numbers are O.D. values; bold numbers indicate specific reactivity based on value above cutoff which was defined as the mean binding of mAb 1418 (specific to parvovirus B19) +3 standard deviations. 2Flow cytometry was used to measure the mAb binding to JRFL, SF162 Env-transfected 293T cells and native 293T cells as control. The bold numbers indicate percent of cells specifically reactive with mAbs based on values above cutoff determined with irrelevant mAb 1418 as described above. C(+) – positive control; C(-) – negative control; nt – not tested.