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Olof Gidlöf

Research project participant

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Quantitation of 87 Proteins by nLC-MRM/MS in Human Plasma : Workflow for Large-Scale Analysis of Biobank Samples

Author

  • Melinda Rezeli
  • Karin Sjödin
  • Henrik Lindberg
  • Olof Gidlöf
  • Bertil Lindahl
  • Tomas Jernberg
  • Jonas Spaak
  • David Erlinge
  • György Marko-Varga

Summary, in English

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 < 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.

Department/s

  • Clinical Protein Science and Imaging
  • Molecular Cardiology

Publishing year

2017-09-01

Language

English

Pages

3242-3254

Publication/Series

Journal of Proteome Research

Volume

16

Issue

9

Document type

Journal article

Publisher

The American Chemical Society (ACS)

Topic

  • Cell and Molecular Biology

Keywords

  • human plasma
  • isotope labeled standards
  • large-scale analysis
  • multiple reaction monitoring
  • protein quantitation

Status

Published

Research group

  • Clinical Protein Science and Imaging
  • Molecular Cardiology

ISBN/ISSN/Other

  • ISSN: 1535-3893