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. 2010 May 23;23(7):567–577. doi: 10.1093/protein/gzq029

Fig. 2.

Fig. 2

Screening of scFvs against (A) biotinylated whole-cell RBE4 lysates or (B) selectively biotinylated plasma membrane proteins residing in RBE4 cell lysates by FACS. Flow cytometric density plots illustrating scFv clone enrichment are shown after each indicated screening round with gates used for quantitative population analysis drawn in. Yeast cells that show both scFv expression (x-axis, detected using an anti-c-myc antibody) and lysate binding (y-axis, detected using SA-PE) were collected in each round. Density plots from TX screens are shown. The percentages of antibody-displaying yeast cells that bind to biotinylated lysate proteins are noted in each density plot for each detergent tested. (C) Assessment of cell lysate binding of individual clones by flow cytometry. Representative density plots of individual clones identified from either whole-cell screening (2T5, 2O1) or plasma membrane focused screening (3mO11) are shown. An anti-hen egg lysozyme scFv D1.3 was used as a negative control. The quantitative binding data for all individual clones are quantified in Table I.