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Technical evaluation of MALDI-TOF mass spectrometry for quantitative proteomic profiling matrix formulation and application

Clinical Proteomics volume 1, pages 259–270 (2004)Cite this article

Abstract

Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS) has been recently used to identify disease markers by directly profiling and quantifying the peptide/proteins in biological samples under different physiological or experimental conditions. The information of reproducibility of such quantitative profiling method has not been available. It is important to evaluate and reduce error from technical variation. In this study, an unbiased signal acquisition strategy was used to evaluate the effects of three sample-matrix spotting methods and two matrix chemicals, α-cyano-4-hydroxycinnamic acid (CHCA) and sinapinic acid, on the reproducibility of the peptide/protein signal intensities. The sandwich spotting method using 0.1% nitrocellulose coating film and CHCA gave the best quantitative results for the standard peptides and proteins with mass<66.5 kDa. The normalized signal intensities of the standard peptides and proteins were directly proportional to their concentrations with intra-assay (within-day) coefficient of variations (CVs) ranging from 6.5% to 17%. When analyzing serum peptides <6000 m/z, the interassay (between-days) CVs of all the evaluated peptide peaks were <15%. These data indicate that with the right MS analysis conditions, MALDI-TOF MS appears to be a feasible tool for directly profiling and quantifying the peptide/ proteins in biological samples.

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Authors and Affiliations

  1. Department of Clinical Oncology, the Sir Y. K. Pao Center for Cancer, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, the People's Republic of China

    Charles M. L. Chan, Ebenezer K. C. Kong & Anthony T. C. Chan

  2. Cancer Research UK Institute for Cancer Studies, The University of Birmingham, Edgbaston, Birmingham, England

    Philip J. Johnson

  3. Department of Medicine and Therapeutics, Chinese University of Hong Kong, Prince of Wales Hospital, Shatin, Hong Kong

    Ronald T. K. Pang, Joseph J. Y. Sung &  Terence C. W. Poon

Authors
  1. Ronald T. K. Pang
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  2. Philip J. Johnson
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  3. Charles M. L. Chan
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  4. Ebenezer K. C. Kong
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  5. Anthony T. C. Chan
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  6. Joseph J. Y. Sung
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  7. Terence C. W. Poon
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Corresponding author

Correspondence to Terence C. W. Poon.

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Pang, R.T.K., Johnson, P.J., Chan, C.M.L. et al. Technical evaluation of MALDI-TOF mass spectrometry for quantitative proteomic profiling matrix formulation and application . Clin Proteom 1, 259–270 (2004). https://doi.org/10.1385/CP:1:3-4:259

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Clinical Proteomics

ISSN: 1559-0275