Figure 1.
Display of human IgG1 Fc on the CHO cell surface.A, schematic of the Fc CHO display construct and the staining strategy. The human IgG1 hinge CH2 and CH3 regions were appended with an N-terminal murine IgK secretory leader sequence (LS), C-terminal (Gly3Ser)2 linker (GS), and PDGFR transmembrane domain and introduced into the pPyEBV vector. The wild-type human IgG1 Fc (WT), an Fc variant with impaired binding to FcγRIIIa (LALAPG), and an Fc variant with enhanced binding to FcγRIIIa (SDALIE) were transfected into CHO cells, stained, and assayed for (B), Fc display level with anti-human Fc-Alexa Fluor 647 (AF647) and (C), binding to biotinylated FcγRIIIa (allele V158) monomer conjugated to streptavidin-PE via flow cytometry. Untransfected controls are also shown; the data are representative of three experimental repeats. CHO, chinese hamster ovary.