Overview of the glycoengineered FcαRI BsAbs. (A) Schematic overview of the developed IgG1-Fc glycovariants, including WT, low fucose, high galactose and combination of high galactose with low fucose. FcαRI BsAbs is a bispecific human IgG1 antibody that targets anti-EGFR and anti-FcαRI. Also, the CH3 domain anti-EGFR G1m(f) and CH3 domain anti-FcαRI G1m(a) shows the heterodimer interaction. Mutation F405L and K409R are indicated in the figure. (B) The percentage of IgG1-Fc fucosylation, IgG1-Fc galactosylation, IgG1-Fc bisection and IgG1-Fc sialyation of the glycovariants is shown in percentage (%). (C) SDS-page of the parentals and glycoengineered FcαRI-BsAbs. Left blot is the 10% non-reduced gel and right blot is the 10% reduced gel. As shown in the blot from l.t.r. the ladder, (1) Cetuximab, (2) anti-FcαRI IgG1, ladder, (3) WT, (4) low fucose, (5) high galactose, (6) the combination and the ladder. Heavy chain (H-chain) and light chain (L-chain) are indicated by black arrows on the right. Created with BioRender.com.