(1061-B) Cell-based metabolic profiling assays for cell therapy.

Cell therapy is an emerging field that utilizes cells as a form of intervention for various diseases and conditions. When developing these therapies, cell health, including energy metabolism, must be considered to elicit optimal results and performance. Ideally, assays that are easy-to-use and high throughput in nature will be beneficial when designing and testing various cells for therapeutic purposes, i.e., CAR T cells. We developed a suite of bioluminescent cell-based assays using NAD(P)H detection technology coupled to dehydrogenases, as well as measuring pyruvate utilizing hydrogen peroxide detection system coupled to oxidase activity. These assays can measure metabolites in key pathways that can be utilized in a high throughput fashion and requires minimal cells for testing. Output of these various assay can address important features of cell health including cell viability, various fuel consumptions, and overall metabolic state of the cell. For example, we have developed assays that measure key fuels, such as glucose, pyruvate, and fatty acids. Along with measuring key metabolites, we have designed multiple assays that measure enzymatic activity for various pathways including glycolysis, the Krebs cycle, and fatty acid oxidation. Further, depending on the metabolite levels in cells, most assays can be utilized with less than 500 cells per well in a 384-well low volume plate, allowing for vast metabolic screening of multiple pathways in about an hour. This format will allow for various research areas to use these assays for an in depth look at overall cell fitness in a convenient and rapid manner.