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

Rezeli, Melinda LU ; Sjödin, Karin LU ; Lindberg, Henrik LU ; Gidlöf, Olof LU ; Lindahl, Bertil; Jernberg, Tomas; Spaak, Jonas; Erlinge, David LU and Marko-Varga, György LU (2017) In Journal of Proteome Research 16(9). p.3242-3254
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... (More)

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.

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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
human plasma, isotope labeled standards, large-scale analysis, multiple reaction monitoring, protein quantitation
in
Journal of Proteome Research
volume
16
issue
9
pages
13 pages
publisher
The American Chemical Society
external identifiers
  • scopus:85028776551
  • wos:000410003500013
ISSN
1535-3893
DOI
10.1021/acs.jproteome.7b00235
language
English
LU publication?
yes
id
2070d7c8-c9de-488e-82c2-9f57b5d7c460
date added to LUP
2017-09-26 13:19:22
date last changed
2018-01-16 13:26:46
@article{2070d7c8-c9de-488e-82c2-9f57b5d7c460,
  abstract     = {<p>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 &lt; 10e-5) for the six proteins, in addition for the cardiovascular predictor factor, APOB: APOA1 ratio (r = 0.969, p-value &lt; 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.</p>},
  author       = {Rezeli, Melinda and Sjödin, Karin and Lindberg, Henrik and Gidlöf, Olof and Lindahl, Bertil and Jernberg, Tomas and Spaak, Jonas and Erlinge, David and Marko-Varga, György},
  issn         = {1535-3893},
  keyword      = {human plasma,isotope labeled standards,large-scale analysis,multiple reaction monitoring,protein quantitation},
  language     = {eng},
  month        = {09},
  number       = {9},
  pages        = {3242--3254},
  publisher    = {The American Chemical Society},
  series       = {Journal of Proteome Research},
  title        = {Quantitation of 87 Proteins by nLC-MRM/MS in Human Plasma : Workflow for Large-Scale Analysis of Biobank Samples},
  url          = {http://dx.doi.org/10.1021/acs.jproteome.7b00235},
  volume       = {16},
  year         = {2017},
}