Fig. 1.

Generation of human monoclonal antibodies from single B cells. Class switched memory B cells were isolated from individuals and microcultured in activating media to induce differentiation into plasmblast/plasma cells to promote IgG secretion for screening against target antigens. VH and VL genes from B-cell cultures positive for the target were recovered by RT-PCR and cloned into a mammalian expression vector for expression as full-length human IgG1 followed by standard IgG purification via fast protein liquid chromatography. Following QC, recombinant mAbs were assessed for functional activity in vitro and in vivo. V_H: heavy chain variable domain, V_L: light chain variable domain, C_H: heavy chain constant domain, C_L: light chain constant domain