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Comparison of methods to examine the endogenous peptides of fetal calf serum

Clinical Proteomics volume 2, pages 67–89 (2006)Cite this article

Abstract

There is a great desire to relate the patterns of endogenous peptides in blood to human disease and drug response. The best practices for the preparation of blood fluids for analysis are not clear and also relatively few of the peptides in blood have been identified by tandem mass spectrometry. We compared a number of sample preparation methods to extract endogenous peptides including C18 reversed phase, precipitation, and ultrafiltration. We examined the results of these sample preparation methods by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) and liquid chromatography-tandem mass spectrometry (MS/MS) using MALDI-TOF/TOF and electrospray ionization-ion trap. Peptides from solid-phase extraction with C18 in the range of hundreds of femtomoles per spot were detected from the equivalent of 1 μL of serum by MALDI-TOF. We observed endogenous serum peptides from fibrinogen α- and β-chain, complement C3, α-2-HS-glycoprotein, albumin, serine (or cysteine) proteinase inhibitor, factor VIII, plasminogen, immunoglobulin, and other abundant blood proteins. However, we also recorded significant MS/MS spectra from tumor necrosis factor-α-, major histocompatibility complex-, and angiotensin-related peptides, as well as peptides from collagens and other low-abundance proteins. Amino acid substitutions were detected and a phosphorylated peptide was also observed. This is the first time the endogenous peptides of fetal serum have been examined by MS and where peptides from low-abundance proteins, phosphopeptides, and amino acid substitutions were detected.

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Authors and Affiliations

  1. Bruker Daltonics, Inc., Bellerica, MA

    Catherine Stacey, Mike McDonell, Paul Kowalski & Jane Marie Kowalski

  2. Department of Pathology and Laboratory Medicine, Mount Sinai Hospital and Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON, Canada

    Eleftherios P. Diamandis

  3. Department of Chemistry and Biology, Faculty of Engineering and Applied Science, Ryerson University, 350 Victoria Street, M5B 2K3, Toronto, ON, Canada

    Declan Williams, Peihong Zhu, Peter Bowden & John Marshall

  4. Ontario Cancer Biomarker Network, MaRS, 101 College Street, Toronto Ontario, ON, Canada

    Ken Evans, Eleftherios P. Diamandis, K. W. Michael Siu & John Marshall

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  1. Declan Williams
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Correspondence to John Marshall.

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Open Access This article is published under license to BioMed Central Ltd. This is an Open Access article is distributed under the terms of the Creative Commons Attribution License ( https://creativecommons.org/licenses/by/2.0 ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Williams, D., Zhu, P., Bowden, P. et al. Comparison of methods to examine the endogenous peptides of fetal calf serum. Clin Proteom 2, 67–89 (2006). https://doi.org/10.1385/CP:2:1:67

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Clinical Proteomics

ISSN: 1559-0275