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Quantitation of 87 Proteins by nLC-MRM/MS in Human Plasma: Workflow for Large-Scale Analysis of Biobank Samples
Lund Univ, Clin Prot Sci & Imaging, Dept Biomed Engn, BMC D13, SE-22184 Lund, Sweden..
H Lundbeck & Co AS, Dept Drug Metab, DK-2500 Copenhagen, Denmark..
Lund Univ, Clin Prot Sci & Imaging, Dept Biomed Engn, BMC D13, SE-22184 Lund, Sweden..
Lund Univ, Skane Univ Hosp, Dept Cardiol, Clin Sci, SE-22185 Lund, Sweden..
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2017 (English)In: Journal of Proteome Research, ISSN 1535-3893, E-ISSN 1535-3907, Vol. 16, no 9, p. 3242-3254Article in journal (Refereed) Published
Abstract [en]

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 (approximate to 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 < 10e-5) for the six proteins, in addition for the cardiovascular predictor factor, APOB: APOA1 ratio (r = 0.969, p-value < 10e-5). Moreover, we utilized the developed assay for screening of biobank samples from patients with myocardial infarction and performed the comparative analysis of patient groups with STEMI (ST- segment elevation myocardial infarction), NSTEMI (non ST- segment elevation myocardial infarction) and type-2 AMI (type-2 myocardial infarction) patients.

Place, publisher, year, edition, pages
AMER CHEMICAL SOC , 2017. Vol. 16, no 9, p. 3242-3254
Keywords [en]
multiple reaction monitoring, isotope labeled standards, large-scale analysis, human plasma, protein quantitation
National Category
Basic Medicine
Identifiers
URN: urn:nbn:se:uu:diva-335416DOI: 10.1021/acs.jproteome.7b00235ISI: 000410003500013PubMedID: 28738677OAI: oai:DiVA.org:uu-335416DiVA, id: diva2:1163165
Funder
Swedish Foundation for Strategic Research , KF10-0024Swedish Research Council, HOR2020-664339VINNOVA, 2016-04733Available from: 2017-12-06 Created: 2017-12-06 Last updated: 2018-01-13Bibliographically approved

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Lindahl, Bertil

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