FIGURE 2.

The lower limit of quantitation for the DEVO assay. Freshly isolated resting CD4 + T cells from an uninfected donor were stimulated with PHA and irradiated PBMC, then co-cultured with PHA blasts over the course of 19 days. On three separate, but not consecutive days, supernatant from day 8 and 12 were spiked with HIV p24 protein at different dilutions and assayed by Simoa. Bias, total error and % total error was calculated from replicate well values using the following formula: Bias = Average−ActualValue;TotalError = |Bias| + 2×StandardDeviation;%TotalError = TotalErrorActualValue×100. The concentration with <20% total error was determined to be 49.21fg/ml**. This value was used as the DEVO lower limit of quantitation.