Figure 3.

TRIC-based single-dose high-throughput screening

TRIC measurement applied for the exhaustive screening of peptide libraries in solution.

(A) Schematic representation of a single-dose TRIC setup. Two optical systems operate simultaneously below a 384-microtiter plate for increased throughput at a distance of nine wells. Unmodified peptides are typically screened as a duplicate. Binding is detected by calculating the area between signal traces recorded in presence and absence of the probed ligand.

(B and C) Bar graph showing area values for each peptide measured (see Tables S3 and S4 for peptide sequences corresponding to the GlyR β (B) and GABA_AR α3 (C) subunit respectively and area values for each data point). Residues delimiting the peptides on-chip are represented on the x axis. Hits are highlighted in dark red alongside corresponding peptide sequences with the respective binding motifs in bold. Note that the software gratifyingly classified sequences as binders that harbor the structurally resolved bindings sites ⁴²⁰FSIVG⁴²⁴ or ³⁹⁵FNIVG³⁹⁹ (GlyR β and GABA_AR α3 subunit respectively). Values are presented as n=1-6 with corresponding STDEV if applicable.