Quantitation of 87 Proteins by nLC-MRM/MS in Human Plasma: Workflow for Large-Scale Analysis of Biobank Samples.
Melinda RezeliKarin SjödinHenrik LindbergOlof GidlöfBertil LindahlTomas JernbergJonas SpaakDavid ErlingeGyörgy Marko-VargaPublished in: Journal of proteome research (2017)
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.
Keyphrases
- st segment elevation myocardial infarction
- liquid chromatography
- mass spectrometry
- percutaneous coronary intervention
- ms ms
- high resolution mass spectrometry
- tandem mass spectrometry
- high throughput
- acute myocardial infarction
- high performance liquid chromatography
- liquid chromatography tandem mass spectrometry
- simultaneous determination
- cardiovascular disease
- gas chromatography
- end stage renal disease
- solid phase extraction
- acute coronary syndrome
- heart failure
- multiple sclerosis
- st elevation myocardial infarction
- coronary artery disease
- high resolution
- left ventricular
- newly diagnosed
- ejection fraction
- chronic kidney disease
- prognostic factors
- electronic health record
- skeletal muscle
- insulin resistance
- case report