Fig. 2.

Assay qualification plan. Four assays were performed using the frozen PBMC of two RA patients R1 (HLA-DRB1∗04:01) and R2 (HLA-DRB1∗01:01), one assay from one sample each by two analysts on two different days. Each analyst independently thawed frozen vials of each RA sample on day 1 and day 2 and performed assays and flow cytometry analysis separately. Mean, SD, and %CV were calculated for different precision measurements. (A) Intra-assay precision of each assay was measured as %CV_1-4 from individual assay replicates. (B) Intra-assay repeatability was measured as mean %CV (%CV_m) of all four assays of each sample. (C) Intra-day Inter-analyst precision was measured as %CV from the mean value of assays performed by two analysts using the same sample on the same day. (D) Inter-assay precision was calculated from all replicates of all four assays performed from the same sample on both the days.