A rapid and high-throughput T cell immunophenotyping assay for cellular therapy bioprocess using the Cellaca PLX image cytometer

Abstract Cellular therapies such as chimeric antigen receptor (CAR) T or NK cells undergo phenotypic characterization during discovery and development of novel therapies, biomanufacturing, upstream patient cell processing for clinical use. Samples are routinely analyzed via flow cytometry cell, B monocyte surface marker expression. Image systems perform cell-based assays that can be used a convenient alternative to cytometry. Recently, new high-throughput image cytometer, the Cellaca PLX system (Nexcelom, Lawrence, MA) was developed immunophenotyping, transfection/transduction efficiency, health assays. This instrument assess several critical quality attributes (CQAs) identity, viability, other relevant biological functions recommended by International Organization Standardization using ISO Cell Characterization focuses on cellular therapeutic products. Here we demonstrate rapid immunophenotyping viability detection in peripheral blood mononuclear (PBMCs) PLX. PBMCs underwent red (RBC) lysis CD3 enrichment, were stained with Hoechst/CD3/CD4/CD8 Hoechst/CD3/CD8/RubyDead kits measured three different cytometers comparison. Results showed highly comparable populations between suggesting cytometer may provide method immunophenotyping. The proposed streamline workflows from sample staining data analysis, quickly moving precious samples into downstream processes.