Figure 1.

Affinity-maturation of the eAd by random mutagenesis and yeast display selection. (A) FACS selection of yeast-displayed 2g2 mutants. During three selections, yeast cells were stained with concentrations of biotinylated IGF-II at 10 nmol/L (sort1), 5 nmol/L (sort2) and 2.5 nmol/L (sort3), respectively. IGF-II-binding was detected by streptavidin-PE. Antibody expression on yeast was shown with mouse anti-c-myc antibody followed by FITC goat anti-mouse secondary antibody. The 0.1-0.3% cells were selected from sort gates. (B) Purification of soluble s7g1 by the size-exclusion column. (C) SDS-PAGE analysis of purified s7g.