Multiplexed LC/ESI-MRM-MS-based Assay for Identification of Coronary Artery Disease Biomarkers in Human Plasma.

PURPOSE: A highly-multiplexed LC/ESI-MRM-MS based assay was developed for the identification of coronary artery disease (CAD) biomarkers in human plasma.

EXPERIMENTAL DESIGN: The assay was used to measure 107 stable isotope labeled peptide standards and native peptides from 64 putative biomarkers of cardiovascular diseases in tryptic digests of plasma from subjects with (n = 70) and without (n = 45) angiographic evidence of CAD and no subsequent cardiovascular mortality during follow-up.

RESULTS: Extensive computational and statistical analysis revealed 6 plasma proteins associated with CAD, namely apolipoprotein CII, C reactive protein, CD5 antigen-like, fibronectin, inter alpha trypsin inhibitor heavy chain H1, and protein S. The identified proteins were combined into a LASSO-logistic score with high classification performance (cross-validated AUC = 0.74). When combined with a separate score computed from markers currently used in the clinic with similar performance, the AUC increased to 0.84. Similar results were observed in an independent set of subjects (n = 87).

CONCLUSIONS AND CLINICAL RELEVANCE: If externally validated, the assay and identified biomarkers can improve CAD risk stratification. This article is protected by copyright. All rights reserved.