Optimization and evaluation of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) with reversed-phase protein arrays for protein profiling

Manuel Aivado, Dimitrios Spentzos, Gil Alterovitz, Hasan H. Otu, Franck Grall, Aristoteles A.N. Giagounidis, Meghan Wells, Je Yoel Cho, Ulrich Germing, Akos Czibere, Wolf C. Prall, Chris Porter, Marco F. Ramoni, Towia A. Libermann

Research output: Contribution to journalArticle

41 Scopus citations

Abstract

Surface-enhanced laser desorption/ionization (SELDI) time-of-flight mass spectrometry with protein arrays has facilitated the discovery of disease-specific protein profiles in serum. Such results raise hopes that protein profiles may become a powerful diagnostic tool. To this end, reliable and reproducible protein profiles need to be generated from many samples, accurate mass peak heights are necessary, and the experimental variation of the profiles must be known. We adapted the entire processing of protein arrays to a robotics system, thus improving the intra-assay coefficients of variation (CVs) from 45.1% to 27.8% (p<0.001). In addition, we assessed up to 16 technical replicates, and demonstrated that analysis of 2-4 replicates significantly increases the reliability of the protein profiles. A recent report on limited long-term reproducibility seemed to concord with our initial inter-assay CVs, which varied widely and reached up to 56.7%. However, we discovered that the inter-assay CV is strongly dependent on the drying time before application of the matrix molecule. Therefore, we devised a standardized drying process and demonstrated that our optimized SELDI procedure generates reliable and long-term reproducible protein profiles with CVs ranging from 26.7% to 32.6%, depending on the signal-to-noise ratio threshold used.

Original languageEnglish (US)
Pages (from-to)133-140
Number of pages8
JournalClinical Chemistry and Laboratory Medicine
Volume43
Issue number2
DOIs
StatePublished - 2005

Keywords

  • High-throughput
  • Plasma protein profiling
  • Proteomics
  • Serum protein profiling
  • Surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI TOF-MS)

ASJC Scopus subject areas

  • Clinical Biochemistry
  • Biochemistry, medical

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    Aivado, M., Spentzos, D., Alterovitz, G., Otu, H. H., Grall, F., Giagounidis, A. A. N., Wells, M., Cho, J. Y., Germing, U., Czibere, A., Prall, W. C., Porter, C., Ramoni, M. F., & Libermann, T. A. (2005). Optimization and evaluation of surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF MS) with reversed-phase protein arrays for protein profiling. Clinical Chemistry and Laboratory Medicine, 43(2), 133-140. https://doi.org/10.1515/CCLM.2005.022