Purification and functions of mAb CO and mAb COK expressed in <i>Arabidopsis</i> plants.

(A) Representative images depicting the mass production of mAb CO and mAb COK in transgenic Arabidopsis plants. Seeds were sown in a greenhouse and conditioned at 24°C, 30% humidity, 16 h light, and 8 h dark. Photographs were taken seven weeks after sowing homozygous seeds. (B) SDS-PAGE analysis of eluted F1–F3 fractions of purified anti-cancer mAbs obtained from Arabidopsis plants. Lane 1: protein marker; lane 2: anti-EpCAM mAb; lane 3: flow-through; lanes 4–6: mAb CO; and lanes 7–9: mAb COK. Samples (20 μL) were loaded into each well. (C) Surface plasmon resonance analysis was used to confirm the binding activity of mAb CO and mAb COK to an epithelial cell adhesion molecule (EpCAM-Fc). To confirm antigen-antibody binding activity of both mAb CO and mAb COK purified from transgenic Arabidopsis plants, the surface of a GLC sensorchip was immobilized with EpCAM fused to human IgG Fc fragment (EpCAM-Fc) molecules. Anti-EpCAM mAb (300 nM), mAb CO (300 nM), and mAb COK (300 nM) were applied to the sensor chip with a flow rate of 80 μL·min-1 at 25°C, and the response curves shown in the figure were consequently obtained.