Fig. 4.

Isolation and purification of yeast SUMO-binding Adhirons. (A) Phage ELISA of Adhirons from 24 clones incubated in wells containing ySUMO (black) or control (grey) showing the TMB product absorbance at 560 nm after 3 min. incubation. (B) Ad-ySUMOs were expressed in BL21 (DE3) cells and cell lysates were heated to 20, 50, 60, 70, 80, 90 and 100°C for 20 min then 5 µl of cleared lysates was separated by 15% SDS–PAGE and Coomasie stained. (C) Ad-ySUMO purification by Ni-NTA beads and analysis of the purified Ad-ySUMOs by 15% SDS–PAGE with Coomassie staining. (D) DSC of Ad-ySUMO clones 10, 15, 20 and 22 together with the Adhiron scaffold.