Comparison of dual bivalent BsAb designs
(A) Schematic of dual bivalent BsAb panel: IgG-[L]-scFv (2+2), BiTE-Fc
(2+2B), and IgG-[H]-scFv (2+2HC). Orange represents anti-huCD3ε domains
(derived from huOKT3) and blue represents anti-GD2 domains (derived from hu3F8).
(B) Representative cell-binding activity of each BsAb against GD2(+) human M14
melanoma cells (left) and CD3(+) activated human T cells (right), measured by
flow cytometry. Geometric mean intensity was normalized to 2+2 (100%) for each
BsAb. (C) Representative T cell-dependent cytotoxicity for each BsAb. (D)
Schematic of in vitro co-culture assay (left) and graph of results (right). For
reference: 2+2 is purple, 2+2B is blue, and 2+2HC is green. Each curve
represents one BsAb, and each point represents a single concentration, with two
(FACS, cytokine) or three (cytotoxicity) technical replicates. Data are shown as
means ± standard deviation.