Characterization of human plasma lipoproteins using anion exchange fast protein liquid chromatography and targeted mass spectrometry assay

Claudia Seidl, Zachary Flaten, Danni Li

Research output: Contribution to journalArticlepeer-review

Abstract

We described a targeted mass spectrometry assay based on selected reaction monitoring (SRM) for five apolipoproteins (apoA1, apoB, apoJ, apoD, and apoE) in plasma lipoproteins isolated by anion exchange fast protein liquid chromatography using only 100 μL of plasma. We performed analytical characterization of the SRM assay and evaluated reproducibility of the entire workflow. The limit of detections for apoA1, apoB, apoD, apoJ, and apoE were 0.6, 4.6, 0.8, 1.2, and 0.7 nM, respectively; the limit of quantifications was 8.3 nM for all peptides except apoD (4.2 nM). The SRM assay was linear from 0.4 to 1667 nM. The intra-day and inter-day and total repeatability (CV%) of the assay ranged from 2.2% to 21.7% for all five peptides. The intra-day and inter-day and total reproducibility of the entire workflow ranged from 12.2% to 43.9% for all five peptides in fractionated high-density lipoprotein, low-density lipoprotein, and IDL. In the future, we will apply this workflow to investigate the association of fractionated plasma lipoproteins with amyloid deposition and cognitive changes in the context of Alzheimer's disease.

Original languageEnglish (US)
Article number2000224
JournalProteomics
Volume21
Issue number6
DOIs
StatePublished - Mar 2021

Bibliographical note

Funding Information:
Research reported in this publication is supported by the NIH/NIA grants R21AG059068, R01AG059654, and R21AG061372. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Publisher Copyright:
© 2021 Wiley-VCH GmbH

Keywords

  • Alzheimer's disease
  • CPTAC
  • SRM
  • plasma lipoproteins
  • proteomics

Fingerprint Dive into the research topics of 'Characterization of human plasma lipoproteins using anion exchange fast protein liquid chromatography and targeted mass spectrometry assay'. Together they form a unique fingerprint.

Cite this