Extended Data Fig. 2: Flow cytometry analysis of library activity on LMVGG over the course of evolution.
Stocks of yeast libraries remaining at the end of each round (R) were regrown, induced, and subjected to cell surface sortase reactions.
Each library was incubated for 1 hour with 1 μM Btn-LMVGG. All libraries were assayed for LMVGG activity in TBS, while the libraries that survived negative selection against fetuin A and other plasma proteins (R9–16) were also assayed in human plasma. Data shown are the mean ± standard deviation of three replicates. Activity:expression is the transpeptidation activity (PE) to sortase expression (FITC) ratio. Overall, the trend towards improved LMVGG activity per unit expression is strong, indicative of a successful evolution campaign.