PMID: 16528911Mar 15, 2006Paper

Quantitative determination of peptides using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

BioTechniques
Jesus A GutierrezJohn E Hale

Abstract

A method is described for the quantitative determination of peptides using matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Known limitations imposed by crystal heterogeneity, peptide ionization differences, data handling, and protein quantification with MALDI-TOF mass spectrometry are addressed in this method with a "seed crystal" protocol for analyte-matrix formation, the use of internal protein standards, and a software package called maldi_quant. The seed crystal protocol, a new variation of the fast-evaporation method, minimizes crystal heterogeneity and allows for consistent collection of protein spectra. The software maldi_quant permits rapid and automated analysis of peak intensity data, normalization of peak intensities to internal standards, and peak intensity deconvolution and estimation for vicinal peaks. Using insulin proteins in a background of other unrelated peptides, this method shows an overall coefficient of variance of 4.4%, and a quantitative working range of 0.58-37.5 ng bovine insulin per spot. Coupling of this methodology to powerful analytical procedures such as immunoprecipitation is likely to lead to the rapid and reliable quantification of biologically relevan...Continue Reading

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Citations

Apr 30, 2008·Proceedings of the National Academy of Sciences of the United States of America·Jesus A GutierrezJohn E Hale
Dec 25, 2008·Briefings in Functional Genomics & Proteomics·Mark W DuncanStephen W Hunsucker
Jan 10, 2008·Journal of Proteome Research·Steven W TaylorSoumitra S Ghosh
Jul 26, 2008·Molecular & Cellular Proteomics : MCP·Emília SzájliKatalin F Medzihradszky
Dec 14, 2017·European Journal of Mass Spectrometry·Stanislav PekovEugene Nikolaev
Apr 12, 2007·Expert Review of Proteomics·Bradley L Ackermann, Michael J Berna
Jun 27, 2007·Proceedings of the National Academy of Sciences of the United States of America·Sang Soo HahPaul T Henderson
Jan 20, 2012·Journal of Proteome Research·N Leigh AndersonDetlef Suckau
Nov 21, 2018·Biomacromolecules·Kyle M KossLarry D Unsworth

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