title: The application of multiple reaction monitoring and multi-analyte profiling to HDL proteins.
abstract: BACKGROUND
  HDL 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).
  
  
  METHODS
  We 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.
  
  
  RESULTS
  We 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.
  
  
  CONCLUSIONS
  MAP 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.
authors.affiliation: Department of Medicine, University of Southern California, Los Angeles, CA, USA. hyassine@usc.edu.
authors.forename: Hussein N
authors.initials: HN
authors.lastname: Yassine
authors.forename: Angela M
authors.initials: AM
authors.lastname: Jackson
authors.forename: Chad R
authors.initials: CR
authors.lastname: Borges
authors.forename: Dean
authors.initials: D
authors.lastname: Billheimer
authors.forename: Hyunwook
authors.initials: H
authors.lastname: Koh
authors.forename: Derek
authors.initials: D
authors.lastname: Smith
authors.forename: Peter
authors.initials: P
authors.lastname: Reaven
authors.forename: Serrine S
authors.initials: SS
authors.lastname: Lau
authors.forename: Christoph H
authors.initials: CH
authors.lastname: Borchers
mesh_terms: D000368:Aged; D001798:Blood Proteins / Q000378:metabolism; D002318:Cardiovascular Diseases / Q000097:blood; D016022:Case-Control Studies; D003920:Diabetes Mellitus / Q000097:blood; D005260:Female; D006801:Humans; D008075:Lipoproteins, HDL / Q000097:blood*; D008297:Male; D008875:Middle Aged; D020543:Proteome / Q000378:metabolism; D018709:Statistics, Nonparametric; D053719:Tandem Mass Spectrometry
chemical_list: D001798:Blood Proteins; D008075:Lipoproteins, HDL; D020543:Proteome