Evaluation of division-arrested cells for cell-based high-throughput screening and profiling.
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Digan, Mary, Pou, Chantevy, Niu, Honglin and Zhang, Ji (2005) Evaluation of division-arrested cells for cell-based high-throughput screening and profiling. Journal of biomolecular screening : the official journal of the Society for Biomolecular Screening, 10 (6). pp. 615-623. ISSN 1087-0571
Abstract
Just-in-time cell supply for cell-based high-throughput screening (HTS) is frequently problematic. In addition to scheduling and logistical issues, quality issues and variability due to passage effect, cell cycle, or confluency contribute to day-to-day signal variability in the course of cell-based HTS campaigns. Cell division-arrest and cryopreservation technologies permit the use of cells as assay-ready reagents for HTS and other cell-based profiling and structure-activity studies. In this report, the authors compare division-arrested and dividing cells in 2 assay types that are dependent on movement of proteins within or through cell membranes: a receptor tyrosine kinase assay involving A431 cells responsive to epidermal growth factor, and a secretion reporter assay, which measures secretion of a reporter gene, secreted alkaline phosphatase. In both assays, dividing and division-arrested cells yielded similar basal and maximal signals at a given cell density. Similar IC50s were obtained for reference inhibitors in each assay, type in both dividing and division-arrested cells. In addition, for the secretion reporter assay, when comparing IC50s obtained from 44 compounds randomly chosen from a primary screening hit list, the rank order of potency obtained from dividing cells and division-arrested cells was essentially identical. Furthermore, the results show that, under certain assay conditions, data generated using division-arrested cells are less variable than those generated using dividing cells. In summary, the results suggest that, in many cases, division-arrested cells can substitute for dividing cells and offer certain advantages for cell-based assays.
| Item Type: | Article |
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| Additional Information: | Publisher's version/PDF cannot be used |
| Keywords: | high-throughput screening; cell-based assay; receptor tyrosine kinase; division-arrest; cryopreservation; secretion assay; assay comparison |
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| Date Deposited: | 14 Dec 2009 14:07 |
| Last Modified: | 31 Jan 2013 01:32 |
| URI: | https://oak.novartis.com/id/eprint/29 |