Fig. 2: Deep screening of a yeast-display-pre-selected VHH library.
a, Workflow of VHH yeast display selections. b, Library statistics, showing the total number of clusters or reads, the number of barcodes or UMIs with 12 replicates and number of unique CDR combinations in the protein space. c, Abundance versus deep-screening (DS) FImean of unique CDRs at 300 nM HEL from the R3 MACS and R3 FACS libraries. The library mean intensity is shown as a grey dashed line, and a solid green line shows the hit threshold of 2× the library background. rS values of 0.361 and 0.442, respectively, show a poor correlation between abundance and deep-screening binding intensities. d, Deep-screening equilibrium-binding and kinetic dissociation curves for clones M5, M6, M14 and M15. Error bars are s.e.m. and n ≥ 12 technical replicates of a given UMI. e, BLI kinetics at 50 nM of the same 4 clones against a HEL–biotin-loaded SA tip. The grey dashed line is denoting the separation of the association (left) and dissociation (right) phases collected during BLI kinetics measurements.
