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Anal Chem. 2017 Feb 21;89(4):2440-2448. doi: 10.1021/acs.analchem.6b04368. Epub 2017 Feb 6.

Multiplex High-Throughput Targeted Proteomic Assay To Identify Induced Pluripotent Stem Cells.

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Centre for Translational Omics, UCL Great Ormond Street Institute of Child Health , London, WC1N 1EH, United Kingdom.
Department of Physiology, Anatomy & Genetics, Oxford University , Oxford, OX1 3PT, United Kingdom.
MRC Laboratory for Molecular Cell Biology, University College London , London, WC1E 6BT, United Kingdom.
Oxford Parkinson's Disease Centre, University of Oxford , Oxford, OX1 3QX, United Kingdom.
James Martin Stem Cell Facility, Sir William Dunn School of Pathology, University of Oxford , Oxford, OX1 3RE, United Kingdom.
Janssen Research and Development , Beerse, 2340, Belgium.
Institute of Genetic Medicine, Newcastle University , Newcastle, NE1 3BZ, United Kingdom.
The Weatherall Institute of Molecular Medicine, University of Oxford, John Radcliffe Hospital , Oxford, OX3 9DS, United Kingdom.


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.

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