Table 5.

Assay validation summary.

Assay parameter	Acceptance criteria	Validation results
LOD	The LOD is the 95th percentile distribution limit of mean SFU/well responses after DMSO mock stimulation	15 SFU/well (75 SFU/million PBMC)
Confirmatory cut point	The confirmatory cut point is the 95th percentile distribution limit of peptide/mock response ratios in naïve animals.	1.69
Precision	For responses ≥ 30 SFU/well: Intra-triplicate CV ≤ 30% Intra-assay CV ≤ 30% Inter-assay CV ≤ 50%	Intra-triplicate CV: 2.2–14.3% Intra-assay CV: 5.5–10.9% Inter-assay CV: 9.9–11.3%
Positive control stimulation acceptance range	PHA responses > LOD	All PHA responses were >LOD.
Linearity	Report cell densities through which a linear regression of responses shows an R2 ≥ 0.95.	25,000–200,000 cells/well
Specificity	No positive response to irrelevant HIV control peptides.	Responses for up 2 μg/mL HIV peptides remained < LOD.