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Mass-Spectrometry Based Characterisation of Infant Whole Saliva Peptidome

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Abstract

The objective of the study was to determine optimal conditions for sampling, sample processing and mass-spectrometry based analysis of infants’ salivary peptidome. Saliva was sampled in 3- and 6-month-old infants and peptide extracts were prepared. Various sample pretreatments before profiling by MALDI-ToF were evaluated and peptide identification was undertaken by MALDI-ToF/ToF or nanoLC-ESI-IT tandem MS. A fast and simple protocol (cut-off filtration at 5 kDa) was satisfactory to produce extracts where no proteolysis was detected even when no protease inhibitor was added. Optimal MALDI spectra were generated after purification on C18 tips. Variability of spectra between two samples exceeded that of the technical replicates, validating that the method is suitable to conduct differential studies. Salivary peptides, identified by means of the two complementary mass spectrometry techniques, were fragments of proline-rich proteins and histatins. The fragments originated mainly from the C-terminal protein extremities. Indications on the proteolytic systems involved and the anatomic location where they intervene are proposed.

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Abbreviations

CID:

Collision-induced dissociation

ESI:

Electrospray ionisation

HPLC:

High pressure liquid chromatography

IT:

Ion trap

LC:

Liquid chromatography

LMW:

Low molecular weight

MALDI:

Matrix-assisted laser description ionisation

MS:

Mass spectrometry

PRPs:

Proline-rich proteins

PSD:

Post source decay

SELDI:

Surface enhanced laser description ionisation

Tof:

Time of flight

WCX:

Weak cation exchange

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Acknowledgments

The authors wish to thank Caroline Laval for participants’ recruitment, Emilie Szleper and Valérie Feyen for saliva sampling, Olivier Palicki for technical assistance for sample preparation and the Opaline team for cohort management. Opaline is approved by Vitagora® (The Taste, Nutrition and Health Innovation Pole). The Opaline project, from which samples were obtained, is financially supported by the Regional Council of Burgundy, IFR92, PRNH INRA-INSERM, ANR-PNRA 2006.

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Authors and Affiliations

  1. CLIPP « Clinical Innovation Proteomic Platform », IFR 100 Santé-STIC, Université de Bourgogne, CHU Le Bocage, 21000, Dijon, France

    G. Lucchi, C. Truntzer, D. Pecqueur & P. Ducoroy

  2. INRA, Plate-forme « Exploration du métabolisme: des gènes aux métabolites », 63122, St Genès Champanelle, France

    C. Chambon

  3. INRA, UMR 1129 FLAVIC, 17 rue Sully, 21000, Dijon, France

    C. Schwartz, S. Nicklaus & M. Morzel

  4. ENESAD, UMR 1129 FLAVIC, 21000, Dijon, France

    C. Schwartz, S. Nicklaus & M. Morzel

  5. Université de Bourgogne, UMR 1129 FLAVIC, 21000, Dijon, France

    C. Schwartz, S. Nicklaus & M. Morzel

Authors
  1. G. Lucchi
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  2. C. Chambon
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  3. C. Truntzer
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  4. D. Pecqueur
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  5. P. Ducoroy
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  6. C. Schwartz
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  7. S. Nicklaus
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  8. M. Morzel
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Corresponding author

Correspondence to M. Morzel.

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Lucchi, G., Chambon, C., Truntzer, C. et al. Mass-Spectrometry Based Characterisation of Infant Whole Saliva Peptidome. Int J Pept Res Ther 15, 177–185 (2009). https://doi.org/10.1007/s10989-009-9167-2

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  • DOI: https://doi.org/10.1007/s10989-009-9167-2

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