Fig. 2

Binding properties of M802. (a) Kinetics of the binding of Herceptin and M802 to immobilized HER2 antigen and of L2K and M802 to immobilized CD3 antigen by SPR assay. M802 (0 to 50 nM) and other antibodies were injected over immobilized HER2 and CD3. (a) and (b) binding curves were analyzed using 1:1 L interaction model. (c) and (d) binding curves were analyzed using Two state interaction model. (b) Dual-binding activity of M802. The HER2 and CD3 antigens were vertically immobilized on two different channels (L4 and L5) in a GLC chip, and then 200 nM M802 was horizontally injected into channels A1, A2, and A3. Then 200 nM CD3 was injected into channel A1, 200 nM HER2 into channel A2, and PBST buffer into channel A3. (c) Cell binding activity of M802. (a) The affinity of M802 at the anti-HER2 moiety on SK-BR-3 cells. (b) The affinity of M802 at the anti-CD3 moiety on the Jurkat cells. (d) M802-mediated cell-to-cell association. The association of SK-BR-3 cells (stained with CFSE) with Jurkat cells (stained with PKH26) without M802 (a) or with M802 (b) was presented. Cells in the upper right quadrant of FL1 vs. FL2 scatter plot represented the double-positive population. (c) The dose dependence of association mediated by M802