Challenges in mass spectrometry-based quantification of bioactive peptides: A case study exploring the neuropeptide Y family

Xi, Li, Jin, Yaping, Parker, Edward A., Josh, Peter, Jones, Alun, Wijffels, Gene, & Colgrave, Michelle L. (2012) Challenges in mass spectrometry-based quantification of bioactive peptides: A case study exploring the neuropeptide Y family. Biopolymers (Chemical Biology) : Peptide Science, 98(4), pp. 357-366.

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The study of biologically active peptides is critical to the understanding of physiological pathways, especially those involved in the development of disease. Historically, the measurement of biologically active endogenous peptides has been undertaken by radioimmunoassay, a highly sensitive and robust technique that permits the detection of physiological concentrations in different biofluid and tissue extracts. Over recent years, a range of mass spectrometric approaches have been applied to peptide quantification with limited degrees of success. Neuropeptide Y (NPY), peptide YY (PYY), and pancreatic polypeptide (PP) belong to the NPY family exhibiting regulatory effects on appetite and feeding behavior. The physiological significance of these peptides depends on their molecular forms and in vivo concentrations systemically and at local sites within tissues. In this report, we describe an approach for quantification of individual peptides within mixtures using high-performance liquid chromatography electrospray ionization tandem mass spectrometry analysis of the NPY family peptides. Aspects of quantification including sample preparation, the use of matrix-matched calibration curves, and internal standards will be discussed. This method for the simultaneous determination of NPY, PYY, and PP was accurate and reproducible but lacks the sensitivity required for measurement of their endogenous concentration in plasma. The advantages of mass spectrometric quantification will be discussed alongside the current obstacles and challenges. © 2012 Wiley Periodicals, Inc. Biopolymers (Pept Sci) 98: 357–366, 2012.

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ID Code: 52019
Item Type: Journal Article
Refereed: Yes
Keywords: Neuropeptide Y, pancreatic polypeptide, peptide YY, multiple reaction monitoring, quantification
DOI: 10.1002/bip.22109
ISSN: 0006-3525
Divisions: Current > Schools > School of Biomedical Sciences
Current > QUT Faculties and Divisions > Faculty of Health
Current > Institutes > Institute of Health and Biomedical Innovation
Copyright Owner: Copyright 2012 John Wiley & Sons Inc
Deposited On: 18 Jul 2012 22:10
Last Modified: 12 Jun 2013 14:58

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