Fig. 1

Bispecific antibody design, purification and SDS-PAGE analysis. A Left panel: Schematic of iMab-N6 engineered using knob-in-a-hole mutations in the CH3 region of N6 and CrossMAb^CH1_CL mutations in iMab. These mutations promote preferential assembly of iMab-N6 by limiting the formation of by-products. A Right panel: To show that these mutations do not affect the function of the antibody, iMab monoclonal antibody was engineered with knob-in-a-hole and CrossMAb^CH1_CL mutations. B Chromatograms of size exclusion chromatography (SEC) purified antibodies with elution fractions corresponding to the correctly assembled antibody confirmation represented by the shaded rectangles. C Antibodies were resolved on a gradient SDS-PAGE post SEC under reduced and non-reduced conditions