Expression and binding characteristics of 5.2-OKT3 biscFv.
(A) Samples of scFv were purified from culture supernatants of recombinant baculovirus-infected cells by a Ni-NTA column as described in “Materials and methods.” Purified 5.2 scFv (lane 1), OKT3 scFv (lane 2), and 5.2-OKT3 biscFv (lane 3) were separated by 10% SDS-PAGE and stained with Coomassie Brilliant Blue R-250 (Nippon Bio-Rad Laboratories, Tokyo, Japan). (B) Native PfMSP-1 (lanes 1-2) and rPfMSP-142 (lanes 3-4) were prepared for SDS-PAGE under nonreducing conditions and run on a 5% to 15% gradient gel. Proteins were transferred to Immobilon membranes (Millipore), probed with either 5.2 mAb followed by biotinylated goat anti–mouse IgG antibody (lanes 1,3) or 5.2-OKT3 biscFv followed by biotinylated mouse anti-FLAG M2 mAb (lanes 2,4).
