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Proteomes 2017, 5(4), 31; doi:10.3390/proteomes5040031

Quantification of Cardiovascular Disease Biomarkers in Human Platelets by Targeted Mass Spectrometry

Leibniz-Institut für Analytische Wissenschaften-ISAS-e.V., 44139 Dortmund, Germany
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Received: 3 October 2017 / Revised: 7 November 2017 / Accepted: 13 November 2017 / Published: 15 November 2017
(This article belongs to the Special Issue Proteomic Biomarkers in Human Diseases)
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Abstract

Platelets are known to be key players in thrombosis and hemostasis, contributing to the genesis and progression of cardiovascular diseases. Due to their pivotal role in human physiology and pathology, platelet function is regulated tightly by numerous factors which have either stimulatory or inhibitory effects. A variety of factors, e.g., collagen, fibrinogen, ADP, vWF, thrombin, and thromboxane promote platelet adhesion and aggregation by utilizing multiple intracellular signal cascades. To quantify platelet proteins for this work, a targeted proteomics workflow was applied. In detail, platelets are isolated and lyzed, followed by a tryptic protein digest. Subsequently, a mix of stable isotope-labeled peptides of interesting biomarker proteins in concentrations ranging from 0.1 to 100 fmol is added to 3 μg digest. These peptides are used as an internal calibration curve to accurately quantify endogenous peptides and corresponding proteins in a pooled platelet reference sample by nanoLC-MS/MS with parallel reaction monitoring. In order to assure a valid quantification, limit of detection (LOD) and limit of quantification (LOQ), as well as linear range, were determined. This quantification of platelet activation and proteins by targeted mass spectrometry may enable novel diagnostic strategies in the detection and prevention of cardiovascular diseases. View Full-Text
Keywords: quantitative proteomics; mass spectrometry; parallel reaction monitoring; cardiovascular diseases; platelets quantitative proteomics; mass spectrometry; parallel reaction monitoring; cardiovascular diseases; platelets
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Malchow, S.; Loosse, C.; Sickmann, A.; Lorenz, C. Quantification of Cardiovascular Disease Biomarkers in Human Platelets by Targeted Mass Spectrometry. Proteomes 2017, 5, 31.

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