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. 2021 Jan 1;11(4):1901–1917. doi: 10.7150/thno.51299

Figure 2.

Figure 2

Screening of Fc libraries for variants with enhanced FcγRIIIa binding. (A) Enrichment of cells with the highest binding to FcγRIIIa during three rounds of selection. HEK293T cells were infected with lentiviral Fc variants libraries at low multiplicity of infection. The cells were stained with FITC conjugated anti-FLAG antibody, biotinylated FcγRIIIa and streptavidin-PE and double positively staining cells were sorted and cultured for the next round of selection. (B) The amino acid distribution at each mutation position of the original libraries and the enriched pools after the 3rd round of selection. The relative frequency of each amino acid at each position is represented by the height of the corresponding symbol. (C, D) Binding of Fc variants with the selected substitutions (C) or the combination of substitutions (D) to FcγRIIIaF158 and FcγRIIIaV158 was measured by surface plasmon resonance (SPR). ΔRU is equal to the response unit of Fc variant subtracted by the response unit of wild-type Fc.