Surfactant-aided precipitation/on-pellet-digestion (SOD) procedure provides robust and rapid sample preparation for reproducible, accurate and sensitive LC/MS quantification of therapeutic protein in plasma and tissues

Analytical Chemistry
Bo AnJun Qu

Abstract

For targeted protein quantification by liquid chromatography mass spectrometry (LC/MS), an optimal approach for efficient, robust and hi-throughput sample preparation is critical, but often remains elusive. Here we describe a straightforward surfactant-aided-precipitation/on-pellet-digestion (SOD) strategy that provides effective sample cleanup and enables high and constant peptide yields in various matrices, allowing reproducible, accurate and sensitive protein quantification. This strategy was developed using quantification of monocolnocal antibody in tissues and plasma as the model system. Surfactant treatment before precipitation substantially increased peptide recovery and reproducibility from plasma/tissue, likely because surfactant permits extensive denaturation/reduction/alkylation of proteins and inactivation of endogenous protease inhibitors, and facilitates removal of matrix components. The subsequent precipitation procedure effectively eliminates the surfactant and nonprotein matrix components, and the thorough denaturation by both surfactant and precipitation enabled very rapid on-pellet-digestion (45 min at 37 °C) with high peptide recovery. The performance of SOD was systematically compared against in-solution-di...Continue Reading

References

Mar 16, 1976·European Journal of Biochemistry·D BarritaultD Hayes
Jan 1, 1983·Annual Review of Biochemistry·J Travis, G S Salvesen
Oct 17, 2002·Protein Science : a Publication of the Protein Society·Birendra N PramanikAjay K Bose
Dec 12, 2002·Chemical Reviews·Peter G W Gettins
May 29, 2004·Analytical Chemistry·Jan Havlis, Andrej Shevchenko
Mar 12, 2005·Proteomics·Linda L ManzaDaniel C Liebler
Sep 13, 2005·Rapid Communications in Mass Spectrometry : RCM·Jun Qu, Robert M Straubinger
Apr 5, 2007·Nature Protocols·Andrej ShevchenkoMatthias Mann
Dec 25, 2008·Journal of Proteome Research·Sheng PanTeresa A Brentnall
Oct 31, 2009·Nature Methods·Daniel C Liebler, Amy-Joan L Ham
Jun 10, 2011·Proteomics·Michael S BeremanMichael J MacCoss
Jun 18, 2011·Bioanalysis·Matthew Ewles, Lee Goodwin
Aug 30, 2011·Journal of Pharmaceutical and Biomedical Analysis·Hanne Kolsrud HustoftHelle Malerod
May 21, 2013·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Irene van den BroekWilliam D van Dongen
Jun 25, 2013·Molecular & Cellular Proteomics : MCP·Ileana R LeónRichard R Sprenger
Aug 28, 2013·Molecular & Cellular Proteomics : MCP·Chengjian TuJun Qu
Sep 11, 2013·Analytica Chimica Acta·Andrew M J CrowellAlan A Doucette
Aug 27, 2014·Bioanalysis·Hendrik NeubertMireia Fernández Ocaña

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Citations

Jun 7, 2016·Clinica Chimica Acta; International Journal of Clinical Chemistry·Sylvain LehmannUNKNOWN IFCC Working Group on Clinical Quantitative Mass Spectrometry Proteomics (WG-cMSP)
Jan 17, 2017·Journal of Proteome Research·Melda Z GürayAlan A Doucette
Apr 22, 2017·Critical Reviews in Biotechnology·Fadzlie Wong Faizal WongDavid C Stuckey
Sep 10, 2017·Molecular & Cellular Proteomics : MCP·Gang LiuSriram Neelamegham
Feb 25, 2018·The Journal of Biological Chemistry·Sabarinath Peruvemba SubramanianRamaswamy Subramanian
Jan 26, 2018·The Journal of Experimental Medicine·Sarah D AckermanKelly R Monk
Jun 26, 2018·Analytical and Bioanalytical Chemistry·Mohsin El AmraniErik M van Maarseveen
May 8, 2018·The Journal of Pharmacology and Experimental Therapeutics·Frank A EnglerJoseph P Balthasar
Jul 2, 2019·Biomedical Chromatography : BMC·Lijuan KangWenying Jian
Jan 30, 2020·Biomedical Chromatography : BMC·Zhendong ChenDafang Zhong
Mar 10, 2018·Frontiers in Pharmacology·Xu ZhuWilliam J Jusko
Apr 1, 2020·American Journal of Physiology. Heart and Circulatory Physiology·Xue WangJun Qu
Apr 13, 2018·Analytical Sciences : the International Journal of the Japan Society for Analytical Chemistry·Kenichiro TodorokiToshimasa Toyo'oka

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