Binding analyses of scFv 183-H12-5C and scFv-Fc 183-H12-5C. Wells of ELISA plate were coated with various dilutions of scFv 183-H12-5C or their parental mAbs. 1 µg/mL of recombinant P24 was captured on immobilized scFv or parental mAbs and detected using HRP-conjugated secondary anti-P24 mAbs (31-90-25). Error bars show standard deviations calculated from at least two independent experiments each done in duplicate (A). Wells of ELISA plate were coated with either 1 µg/mL or 10 µg/mL of recombinant P24. 100 µL of 1ug/mL of scFv-Fc 183-H12-5C was added to capture immobilized recombinant p24 and detected using HRP-conjugated secondary anti-Human. Error bars show standard deviations calculated from at least two independent experiments each done in duplicate (B). Recombinant p24 and proteins extracted from HIV-1 virus were electrophoresed and transferred to nitrocellulose membranes. Immunoblotting with mAb-IgG 31-90-25 or scFv-Fc 183-H12-5C antibodies was carried out as described in materials and methods (C).