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. Author manuscript; available in PMC: 2021 Jul 11.
Published in final edited form as: Nat Chem Biol. 2021 Jan 11;17(3):317–325. doi: 10.1038/s41589-020-00706-1

Extended Data Fig. 2: Flow cytometry analysis of library activity on LMVGG over the course of evolution.

Extended Data Fig. 2:

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.