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This version was published on June 1, 2008
Journal of Biomolecular Screening, Vol. 13, No. 5, 354-362 (2008)
DOI: 10.1177/1087057108317768

Use of Cryopreserved Cells for Enabling Greater Flexibility in Compound Profiling

M.J. Wigglesworth

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK, M.J.Wigglesworth{at}GSK.com

K.J. Lawless

Biopharm Discovery Technology Group

D.J. Standing

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

E.K. Mackenzie

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

V.R. Kitchen

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

F. Mckay

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

E. Ward

Biochemical Cell Targets

S.J. Brough

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

M. Stylianou

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

F.R. Jewitt

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

A.M. Mclaren-Douglas

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

M.I. Jowet

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

N. Tamayama

Biological Reagents and Assay Development, GlaxoSmithKline, Stevenage, Hertfordshire, UK

D. Finnigan

Biological Reagents and Assay Development, GlaxoSmithKline, Stevenage, Hertfordshire, UK

J. Ding

Discovery Statistics, Upper Providence, Collegeville, PA

A. Wise

Screening and Compound Profiling, GlaxoSmithKline, Harlow, UK

Measurement of intracellular calcium release following agonist challenge within cells expressing the relevant membrane protein is a commonly used format to derive structure-activity relationship (SAR) data within a compound profiling assay. The Fluorometric Imaging Plate Reader (FLIPR) has become the gold standard for this purpose. FLIPR traditionally uses cells that are maintained in continuous culture for compound profiling of iterative chemistry campaigns. This supply dictates that assays can only be run on 4 of 5 weekdays, or alternative cell culture machinery is required such that plating can occur remotely at the weekend. The data reported here demonstrate that high-quality compound profiling data can be generated from the use of cryopreserved cells and that these cells can also be plated at various densities to generate equivalent data between 24 and 72 h post-plating. Hence, the authors report a method that allows data generation throughout the week and without the requirement of highly automated cell culture or continuous culture. (Journal of Biomolecular Screening 2008:354-362)

Key Words: cryopreserved • FLIPR • histamine • alpha 1A • alpha 1B


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