TABLE 1.
EASY-HIT assay characteristics according to NIH NCGC guidelines
| Assay parametera | Value for the parameter in: |
NCGC acceptance criterion | |||||
|---|---|---|---|---|---|---|---|
| Validation experimentb |
LOPAC1280 library screeningc |
||||||
| Expt 1 | Expt 2 | Expt 3 | Mean | Step 1 | Step 2 | ||
| CV high (%) | 0.88 ± 0.12 | 1.09 ± 0.30 | 1.12 ± 0.15 | 1.03 ± 0.21 | 2.07 ± 0.59 | 2.06 ± 0.60 | ≤20 |
| CV middle (%) | 1.84 ± 0.30 | 1.68 ± 0.20 | 1.23 ± 0.15 | 1.59 ± 0.34 | ND | ND | |
| CV low (%) | 1.84 ± 0.25 | 1.88 ± 0.25 | 1.61 ± 0.37 | 1.78 ± 0.29 | 11.85 ± 4.01 | 6.00 ± 1.84 | |
| Z′ | 0.97 ± 0.01 | 0.97 ± 0.01 | 0.97 ± 0.00 | 0.97 ± 0.01 | 0.94 ± 0.02 | 0.94 ± 0.02 | 0.4 |
| SWc | 105 ± 7.56 | 86.13 ± 18.61 | 90.83 ± 7.08 | 93.99 ± 13.63 | 49 ± 17 | 50 ± 17 | ≥2 |
| S/Bd (fold) | 95 ± 3 | 62 ± 6 | 57 ± 4 | 71 ± 19 | 157 ± 24 | 330 ± 33 | |
a
CV, coefficient of variation; SW, signal window; S/B, signal-to-background ratio.
b
Values indicated for each experiment represent the mean ± SD of values from three plates, with 60 wells analyzed for each signal. Each experiment was performed on a different day.
c
Forty-eight plates were used for each step. ND, not done.