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Multiplex high-throughput targeted proteomic assay to identify induced pluripotent stem cells

Baud, Anna, Wessely, Frank, Mazzacuva, Francesca, McCormick, James, Camuzeaux, Stephane, Heywood, Wendy E., Little, Daniel, Vowles, Jane, Tuefferd, Marianne, Mosaku, Olukunbi, Lako, Majlinda, Armstrong, Lyle, Webber, Caleb, Cader, M. Zameel, Peeters, Pieter, Gissen, Paul, Cowley, Sally A. and Mills, Kevin 2017. Multiplex high-throughput targeted proteomic assay to identify induced pluripotent stem cells. Analytical Chemistry 89 (4) , pp. 2440-2448. 10.1021/acs.analchem.6b04368

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Abstract

Induced pluripotent stem cells have great potential as a human model system in regenerative medicine, disease modeling, and drug screening. However, their use in medical research is hampered by laborious reprogramming procedures that yield low numbers of induced pluripotent stem cells. For further applications in research, only the best, competent clones should be used. The standard assays for pluripotency are based on genomic approaches, which take up to 1 week to perform and incur significant cost. Therefore, there is a need for a rapid and cost-effective assay able to distinguish between pluripotent and nonpluripotent cells. Here, we describe a novel multiplexed, high-throughput, and sensitive peptide-based multiple reaction monitoring mass spectrometry assay, allowing for the identification and absolute quantitation of multiple core transcription factors and pluripotency markers. This assay provides simpler and high-throughput classification into either pluripotent or nonpluripotent cells in 7 min analysis while being more cost-effective than conventional genomic tests.

Item Type: Article
Date Type: Publication
Status: Published
Schools: Medicine
Publisher: American Chemical Society
ISSN: 0003-2700
Date of Acceptance: 11 January 2017
Last Modified: 21 Oct 2020 14:30
URI: http://orca-mwe.cf.ac.uk/id/eprint/135767

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