Development of high-throughput chemical extraction techniques and quantitative HPLC-MS/MS (SRM) assays for clinically relevant plasma proteins

Journal of Proteome Research
Chris BartonSteve Pleasance

Abstract

The clinical application of targeted plasma protein analysis by selective reaction monitoring of peptides using LC-MS/MS requires the development of robust, inexpensive protein extraction techniques with the potential for high-throughput applications. We present the development of a novel mixed-mode solid phase extraction (SPE) technique for the removal of high abundance and high molecular weight proteins from plasma. This technique, coupled with fused-core HPLC-MS/MS analysis is compared to a previously developed extraction method to study a range of proteins in plasma, including routinely measured biomarkers of growth hormone action. To further validate this technique, it was used for the quantification of insulin-like growth factor I (IGF-I) levels and compared to a state-of-the-art immunoassay on a fully automated analyzer. Clinical reference materials were applied for method development to allow for further interlaboratory comparisons. The LC-MS/MS approach quantified IGF-I in plasma with an accuracy that is within the guidelines for macromolecular assays in a regulated laboratory environment. Furthermore, IGF-I levels determined using the SPE and ACN methods with LC-MS/MS analysis correlated well with the immunoassay resu...Continue Reading

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Citations

Jun 4, 2010·The Analyst·Carol E ParkerChristoph H Borchers
Oct 13, 2012·Molecular & Cellular Proteomics : MCP·Shannon K Flood-NicholsDanielle L Ippolito
Apr 27, 2012·Bioanalysis·Richard G Kay, Andrew Roberts
Dec 19, 2012·Bioanalysis·James W HowardColin S Creaser
Dec 18, 2013·Bioanalysis·Mariëtte Theodora Ackermans, Erik Endert
Oct 14, 2014·Proteomics·Eivind LarssenPeter Ruoff
May 6, 2015·Methods : a Companion to Methods in Enzymology·Hemamalini Ketha, Ravinder J Singh
Apr 15, 2015·Bioanalysis·Daniel WilffertNico C van de Merbel
Oct 18, 2011·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Manfred Rauh
Jun 7, 2016·Clinica Chimica Acta; International Journal of Clinical Chemistry·Sylvain LehmannUNKNOWN IFCC Working Group on Clinical Quantitative Mass Spectrometry Proteomics (WG-cMSP)
Sep 7, 2016·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Abhinav KumarNicole Zitzmann
Jan 4, 2012·Proteomics. Clinical Applications·Christoph KrispMark P Molloy

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