The application of multiple reaction monitoring and multi-analyte profiling to HDL proteins

Lipids in Health and Disease

Research

Derek Smith¹  Chad R Borges¹  Hyunwook Koh²  Hussein N Yassine²  Peter Reaven³  Serrine S Lau⁴  Dean Billheimer⁵  Angela M Jackson⁶  Christoph H Borchers⁷ 

[1] Arizona State University, Tempe, AZ, USA;Department of Medicine, University of Southern California, Los Angeles, CA, USA;Phoenix VA Health Care System, Phoenix, AZ, USA;Southwest Environmental Health Sciences Center, Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, AZ, USA;Statistics Consulting Laboratory, University of Arizona, Tucson, AZ, USA;University of Victoria - Genome British Columbia Proteomics Centre, Victoria, BC, Canada;University of Victoria - Genome British Columbia Proteomics Centre, Victoria, BC, Canada;Department of Biochemistry and Microbiology, University of Victoria, Victoria, BC, Canada;

关键词: High density lipoprotein;    Proteomics;    Multiple reaction monitoring;    Multi-analyte panel;    Diabetes;    Cardiovascular disease;   

DOI  :  10.1186/1476-511X-13-8

 received in 2013-09-26, accepted in 2013-12-26,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundHDL carries a rich protein cargo and examining HDL protein composition promises to improve our understanding of its functions. Conventional mass spectrometry methods can be lengthy and difficult to extend to large populations. In addition, without prior enrichment of the sample, the ability of these methods to detect low abundance proteins is limited. Our objective was to develop a high-throughput approach to examine HDL protein composition applicable to diabetes and cardiovascular disease (CVD).MethodsWe optimized two multiplexed assays to examine HDL proteins using a quantitative immunoassay (Multi-Analyte Profiling- MAP) and mass spectrometric-based quantitative proteomics (Multiple Reaction Monitoring-MRM). We screened HDL proteins using human xMAP (90 protein panel) and MRM (56 protein panel). We extended the application of these two methods to HDL isolated from a group of participants with diabetes and prior cardiovascular events and a group of non-diabetic controls.ResultsWe were able to quantitate 69 HDL proteins using MAP and 32 proteins using MRM. For several common proteins, the use of MRM and MAP was highly correlated (p < 0.01). Using MAP, several low abundance proteins implicated in atherosclerosis and inflammation were found on HDL. On the other hand, MRM allowed the examination of several HDL proteins not available by MAP.ConclusionsMAP and MRM offer a sensitive and high-throughput approach to examine changes in HDL proteins in diabetes and CVD. This approach can be used to measure the presented HDL proteins in large clinical studies.

【 授权许可】

CC BY   
© Yassine et al.; licensee BioMed Central Ltd. 2014