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Quantitation of 87 proteins by nLC-MRM/MS in human plasma: workflow for large-scale analysis of biobank samples Melinda Rezeli, Karin Sjödin, Henrik Lindberg, Olof Gidlöf, Bertil Lindahl, Tomas Jernberg, Jonas Spaak, David Erlinge, and György Marko-Varga J. Proteome Res., Just Accepted Manuscript • DOI: 10.1021/acs.jproteome.7b00235 • Publication Date (Web): 25 Jul 2017 Downloaded from http://pubs.acs.org on July 27, 2017
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Journal of Proteome Research is published by the American Chemical Society. 1155 Sixteenth Street N.W., Washington, DC 20036 Published by American Chemical Society. Copyright © American Chemical Society. However, no copyright claim is made to original U.S. Government works, or works produced by employees of any Commonwealth realm Crown government in the course of their duties.
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Journal of Proteome Research
Quantitation of 87 proteins by nLC-MRM/MS in human plasma: workflow for large-scale analysis of biobank samples Melinda Rezeli1*, Karin Sjödin2, Henrik Lindberg1, Olof Gidlöf3, Bertil Lindahl4, Tomas Jernberg5, Jonas Spaak5, David Erlinge3, György Marko-Varga1 1
Clinical Protein Science & Imaging, Department of Biomedical Engineering, Lund University, Lund, Sweden 2
3
H Lundbeck & Co AS, Department of Drug Metabolism, Copenhagen, Denmark
Department of Cardiology, Clinical Sciences, Lund University, Skåne University Hospital, Lund, Sweden 4
Department of Medical Sciences, Cardiology & Uppsala Clinical Research Center, Uppsala University, Uppsala, Sweden
5
Department of Clinical Sciences, Danderyd University Hospital, Karolinska Institutet, Stockholm, Sweden
Corresponding author: Melinda Rezeli Div. Clinical Protein Science & Imaging, Department of Biomedical Engineering, Lund University, BMC D13, SE-221 84 Lund, Sweden Phone: +46-46-222 3721 Email:
[email protected] ACS Paragon Plus Environment
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Journal of Proteome Research
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Abstract A multiple reaction monitoring (MRM) assay was developed for precise quantitation of 87 plasma proteins including the three isoforms of apolipoprotein E (APOE) associated with cardiovascular diseases using nanoscale liquid chromatography separation and stable isotope dilution strategy. The analytical performance of the assay was evaluated and we found an average technical variation of 4.7 % in 4 - 5 orders of magnitude dynamic range (≈ 0.2 mg/L to 4.5 g/L) from whole plasma digest. Here, we report a complete workflow, including sample processing adapted to 96-well plate format and normalization strategy for large-scale studies. To further investigate the MS-based quantitation the amount of six selected proteins was measured by routinely used clinical chemistry assays as well and the two methods showed excellent correlation with high significance (p-value
