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. 2020 Dec 7;24(1):101898. doi: 10.1016/j.isci.2020.101898

Figure 4.

Figure 4

TRIC-based affinity determination

(A) Schematic representation of a dose response TRIC setup. Two optical systems operate simultaneously below a 384-microtiter plate for increased throughput at a distance of nine wells. Unmodified peptides are analyzed as a twelve-point dilution series with a dilution factor of two. The resulting TRIC traces are then analyzed for Fnorm by comparison of the relative fluorescence at F0 to F1.

(B) Comparison of five independent measurements of a GlyR β-derived peptide (414DLRSNDFSIVGSLPR428) with respective Ki and S/N values. The bright red dot was identified as an outliner and excluded from curve fitting.

(C) Violin plot showing the mean EC50 value of the GlyR β-derived peptides in (B), alongside the mean, median, 25%–75% quantile and range within the 1.5 interquartile range (IQR). The high reproducibility indicates that TRIC-based affinity determination in nanomolar scale is feasible.