Figure 7.

Analysis of Droplet Sorting and Enrichment of OKT 9 Cells by Real-Time PCR

(A) For analyzing the OKT 9 cell enrichment in quantitative terms by real-time PCR, a standard curve was prepared by mixing cells in varying ratios and then comparing their Ct values (Table S3).

(B) Analysis of sorted cell population by real-time PCR showed that the 1:20, 1:100, and 1:400 spike-in ratio for OKT 9 cells before sorting could be enriched to 71%, 49.5%, and 55%, respectively, after FADS. In three separate experiments at 1:400 spike-in ratio (1:400^∗, separated by blue dotted line), the sorted hybridoma cells were recovered from the droplets, cultured and used for another round of sorting. From an average enrichment of 48% of OKT 9 cells in the first round of sorting, the enrichment could be increased to up to 75% in the second round of sorting.

Mean of the percent of OKT 9 cells from 3 independent experiments ± SD is shown.