Fig. 5: DP-DEL PLAP selections.

A DP-DEL HTDS of concatenated triplicate selections against PLAP. Building blocks 1, 2 and 3 correspond to each respective axis. The heat bar represents DNA sequence counts. Plot is constrained to combinations from version 1 of DP-DEL, for both BB4 positions (see supplementary section 5.4.2.). Arrows indicate the 3 highest enriched library members from the selection. Cut-off: 30 counts. B On-LNA experimental validation of highest enriched library members 9-14 displayed on 8-mer locked nucleic acid (LNA) heteroduplexes with a fluorescein label by fluorescence polarization (FP). Conjugates were compared in their version 1 form with an azide group at the BB2 position or an NH₂ group. C Optimization of dual display partners for best performing binders 10 and 12. Two additional enriched BB3 + BB4 combinations as well as a negative control were tested to improve on binders 10 and 12. D Testing of different length maleimide linkers between cysteine scaffolds of both strands, to determine optimal spacing of best performing dual display partners 17. E Off-DNA resynthesis and inhibition experiments of optimized binder 22 as well as its single display variant 21 (R₂ = Ahx-(H-Lys-NH₂)-FITC). Affinity and IC₅₀ values from experimental triplicates (n = 3) are given as a mean. Error bars indicate standard deviation of the replicates.