Table 1. Workflow of high-throughput... | Rockefeller University Press

Table 1.

Workflow of high-throughput primary screening

Step	Test compound concentration	Number of compounds	% of picked compounds	Inhibition cut-off (%)
Total library compound	-	93,716	-	-
Primary assay hits using FP	20 µM	3,010	3.21	>75
Secondary assay hits using AlphaLISA (AL)	12.5 µM	167	0.18	>50
Filtering using quenching (AL)	12.5 µM	97	0.1	<30
Lipinski’s rule		87	0.09	≤1 violation
Validation/dose–response Hits (FP and AL)	0.31–40 µM	26	0.028	IC₅₀ <50 µM
Dose–response using fresh compound (FP and AL)	0.07–40 µM	10	0.011	IC₅₀ <50 µM

Step	Test compound concentration	Number of compounds	% of picked compounds	Inhibition cut-off (%)
Total library compound	-	93,716	-	-
Primary assay hits using FP	20 µM	3,010	3.21	>75
Secondary assay hits using AlphaLISA (AL)	12.5 µM	167	0.18	>50
Filtering using quenching (AL)	12.5 µM	97	0.1	<30
Lipinski’s rule		87	0.09	≤1 violation
Validation/dose–response Hits (FP and AL)	0.31–40 µM	26	0.028	IC₅₀ <50 µM
Dose–response using fresh compound (FP and AL)	0.07–40 µM	10	0.011	IC₅₀ <50 µM