SINGLE-CELL BIOCHEMICAL PROFILING OF LEUKOCYTES FROM WHOLE BLOOD

Abstract

Secreted enzymes and cytokines reflect functionality of leukocytes. Single-cell assay is needed to identify precise cell subsets with high functional activity. A major challenge for measuring single-cell secretion from blood is the time-consuming and unphysiological sample preparation processes. To address this challenge, this interdisciplinary research developed a novel system for 40 min 1-stop assay from whole blood to test single-leukocyte for up to 5 targets of surface antigen, secreted enzyme and cytokine. First, an integrated microfluidic device was developed automates 6 steps of leukocyte extraction, washing, reagent filling, single-cell encapsulation, incubation and profiling. Next, a customized cytometer system was built to enable novel assay for sensitive detection of secreted enzyme and cytokine. Finally, the system was validated by 2 applications: (1) enzyme secretion and surface antigen quantification evaluating inflammation in acute myocardial infarction (AMI) (2) cytokine secretion test for engineered chimeric antigen receptor (CAR) T cell.