Table 3.

HC and LC variants to direct the assembly of anti-HER2/CD3 BsIgG of different heavy chain isotypes and species. Anti-HER2 and anti-CD3 HC and κ-LC variants were co-transfected into Expi293F cells. The resultant BsIgG-containing mixture was purified from the cell culture harvest fluid by protein A chromatography and then analyzed by liquid chromatography-EMR Orbitrap mass spectrometry. Data shown are for optimized LC ratios. Human IgG₁ data are included from Table 1 for comparison. The reverse chimeric BsIgG_2a includes humanized variable domains and mouse κ-LC and mouse IgG_2a HC. See Table 1 legend for additional details.

	BsIgG (%)
Orthogonal pairing variant	Human IgG1	Human IgG2	Human IgG4	Reverse chimeric IgG2a
Parent	24.6	24.4	22.2	21.8
v10	91.7	97.8	90.1	100.0
v11	100.0	100.0	97.6	91.6