Title: Abstract 3643: Flow cytometry multiplexing used to analyze metabolic activity and assess pharmaceutical compound toxicity as a high throughput screening tool
Abstract:Abstract High throughput screening (HTS) is an extremely effective method for allowing researchers to identify putative active compounds for therapeutics. Recently, flow cytometry has emerged as a pow...Abstract High throughput screening (HTS) is an extremely effective method for allowing researchers to identify putative active compounds for therapeutics. Recently, flow cytometry has emerged as a powerful HTS tool with the added benefit of cell-by-cell analysis. Flow cytometry allows a researcher to study protein-protein interactions, metabolic activities, as well as DNA content in a single or multiplexed assay format. For this study, we screened the Tocriscreen Total compound library and compared HTS using a plate reader with flow cytometer multiplexing. Peripheral blood mononuclear (PBMCs) and Jurkat cells were used as cell models. Cells were cultured under standard conditions and either at normoxic (5%) or hyperoxic (19%) oxygen levels. We also varied length of time the cells were treated with compound library (24, 48, or 72 hrs). Membrane integrity and metabolic activity were measured as an output for evaluating cellular viability. To further assess cellular activity, post-screening analysis was implemented to establish EC50s of “hits” from compound library. Screening analysis identified target drugs that were further analyzed using cell health readouts with different mechanisms of action to assess compound toxicity. Results indicate that compound “hits” and potency differed in the screen depending on cell type, the mechanistic readout, length of compound exposure, and mode of readout used to perform the HTS experiments. Citation Format: Mike OGrady, Leticia A. Montoya, Michelle Yan, Scott T. Clarke, Quentin Low, Carolyn DeMarco, April Anderson, Kathy Kihn, Carmen Finnessy, Marcy Wickett, Veronica Calderon. Flow cytometry multiplexing used to analyze metabolic activity and assess pharmaceutical compound toxicity as a high throughput screening tool. [abstract]. In: Proceedings of the 107th Annual Meeting of the American Association for Cancer Research; 2016 Apr 16-20; New Orleans, LA. Philadelphia (PA): AACR; Cancer Res 2016;76(14 Suppl):Abstract nr 3643.Read More
Publication Year: 2016
Publication Date: 2016-07-15
Language: en
Type: article
Indexed In: ['crossref']
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Cited By Count: 1
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