The application of ultra-performance liquid chromatography/tandem mass spectrometry to the detection and quantitation of apolipoproteins in human serum

107Citations
Citations of this article
53Readers
Mendeley users who have this article in their library.
Get full text

Abstract

The detection and quantitation of apolipoproteins, important markers for coronary heart disease, in serum by liquid chromatography/tandem mass spectrometry (LC/MS/MS) using multiple reaction monitoring (MRM) is reported. A tryptic digest of depleted human serum was analysed by nanoflow LC/MS/MS at a flow rate of 300 nL/min and several apolipoproteins (Apo), including Apo A1, A2, A4, C1, C2, C3, D, F and M, were successfully identified. The analysis of the same depleted serum digest by ultra-performance (UP)LC/MS/MS operating at 700 μL/min resulted in comparable sensitivity and selectivity to the nanoflow method, but with a dramatic (∼20-fold) reduction in run time. The potential of UPLC/MS/MS for the rapid quantitation of proteins in biological matrices by representative tryptic peptides was further investigated using Apo A1 and its corresponding stable isotopically labelled tryptic AQUA peptide (DYVSQFEGSALGK). A set of serum-based Apo A1 calibrators from a clinical analyser kit were digested without depletion following the addition of the AQUA peptide and analysed using UPLC/MS/MS. A linear calibration curve was generated from peak area ratios to the labelled peptide with a coefficient of correlation of 0.9989. Standard curves were also generated for other apolipoproteins together with Apo B100, Apo E, lecithin cholesterol acyltransf erase and albumin, which were also detected in the standards. The concentration of Apo A1 in five fresh undepleted human serum samples and a quality control (QC) sample were determined using both the UPLC/MS/MS method and a clinical analyser. Results were comparable and the quantitative study, involving 80 injections which took hours rather than days to complete, demonstrates the high-throughput potential of UPLC/MS/MS to quantify multiple serum proteins without the need for antibodies, and thus provide an alternative to the use of clinical analysers for serum protein biomarkers. Copyright © 2007 John Wiley & Sons, Ltd.

References Powered by Scopus

Quantitative mass spectrometric multiple reaction monitoring assays for major plasma proteins

1115Citations
N/AReaders
Get full text

Shotgun proteomics implicates protease inhibition and complement activation in the antiinflammatory properties of HDL

835Citations
N/AReaders
Get full text

Quantification of C-reactive protein in the serum of patients with rheumatoid arthritis using multiple reaction monitoring mass spectrometry and <sup>13</sup>C-labeled peptide standards

376Citations
N/AReaders
Get full text

Cited by Powered by Scopus

Proteomics by mass spectrometry: Approaches, advances, and applications

867Citations
N/AReaders
Get full text

A review of current trends and advances in modern bio-analytical methods: Chromatography and sample preparation

505Citations
N/AReaders
Get full text

Towards absolute quantification of therapeutic monoclonal antibody in serum by LC-MS/MS using isotope-labeled antibody standard and protein cleavage isotope dilution mass spectrometry

200Citations
N/AReaders
Get full text

Register to see more suggestions

Mendeley helps you to discover research relevant for your work.

Already have an account?

Cite

CITATION STYLE

APA

Kay, R. G., Gregory, B., Grace, P. B., & Pleasance, S. (2007). The application of ultra-performance liquid chromatography/tandem mass spectrometry to the detection and quantitation of apolipoproteins in human serum. Rapid Communications in Mass Spectrometry, 21(16), 2585–2593. https://doi.org/10.1002/rcm.3130

Readers' Seniority

Tooltip

PhD / Post grad / Masters / Doc 21

45%

Researcher 20

43%

Professor / Associate Prof. 5

11%

Lecturer / Post doc 1

2%

Readers' Discipline

Tooltip

Chemistry 20

50%

Agricultural and Biological Sciences 11

28%

Biochemistry, Genetics and Molecular Bi... 5

13%

Medicine and Dentistry 4

10%

Save time finding and organizing research with Mendeley

Sign up for free