Ran across RO testing method used for Montifiore p
Post# of 148177
“” CCR5 receptor occupancy
We determined CCR5 receptor occupancy by leronlimab using phycoerythrin-labeled leronlimab (IncellDx Inc.) in a competitive flow cytometry assay. CCR5-expressing immune cells including CD4+, CD45RO+ T-lymphocytes, CD4+, FoxP3+ T-regulatory cells, and CD14+, CD16+ monocytes/macrophages were included in the panel using the appropriate immunophenotypic markers for each population in addition to phycoerythrin-labeled leronlimab. Cells were incubated for 30 min in the dark at room temperature and washed twice with 2% BSA solution. Flow acquisition was conducted with a three-laser CytoFLEX fitted with 405 nm (80 mW), 488 nm (50 mW), and 638 nm (50 mW) lasers (Beckman Coulter, Indianapolis, IN, USA). Receptor occupancy was determined based on the loss of CCR5 detection over time in the subpopulations, and calculated using the following equation: 1–A/B × 100, where A is day 0 and B is day 7.””