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SELDI-TOF-MS of saliva: methodology and pre-treatment effects.

Author(s): Schipper R, Loof A, de Groot J, Harthoorn L, Dransfield E, van Heerde W

Publication: J Chromatogr B Analyt Technol Biomed Life Sci, 2007, Vol. 847, Page 45-53

PubMed ID: 17070117 PubMed Review Paper? No

Purpose of Paper

The purpose of this paper was to evaluate potential effects of the following preanalytical variables on saliva protein profiles captured by SELDI-TOF mass spectrometry: stimulation prior to saliva collection (chewing), a delay in processing, centrifugation speed, protease inhibitor treatment, storage temperature and duration.

Conclusion of Paper

Although the magnitude of effects was small, the number of saliva protein profile peaks was affected by stimulation prior to collection, while the intensity of peaks was influenced by a 3 h processing delay on ice, centrifugation speed, and storage for 6 months at -20 but not -80 degrees C.

Studies

  1. Study Purpose

    The purpose of this study was to evaluate potential effects of the following preanalytical variables on saliva protein profiles captured by SELDI-TOF mass spectrometry: stimulation prior to saliva collection (chewing), a delay in processing, centrifugation speed (1000 x g versus 10000 x g), protease inhibitor treatment, storage temperature and duration.

    Summary of Findings:

    While collective effects of preanalytical handling variables were small, the number of SELDI-TOF mass spectrometry detected protein peaks varied as a result of stimulation prior to specimen collection. Peak intensity was altered as a result of a 3 h delay on ice prior to processing, and centrifugation speed. Stimulated and non-stimulated saliva specimens yielded similar results with regard to protein stability after 6 months of frozen storage compared to 1 month controls, with reduced intensities at -20 but not -80 degrees C. Treatment with a protease inhibitor cocktail after centrifugation and prior to frozen storage prevented some but not all of the observed profile changes.

    Biospecimens
    Preservative Types
    • Frozen
    Diagnoses:
    • Normal
    Platform:
    AnalyteTechnology Platform
    Protein SELDI-TOF MS
    Pre-analytical Factors:
    ClassificationPre-analytical FactorValue(s)
    Biospecimen Acquisition Type of collection container/solution Salivette collection device
    "Universal collection container"
    Biospecimen Acquisition Method of fluid acquisition Non-stimulated saliva
    Stimulated saliva (chewing prior to collection)
    Storage Storage duration 0 h
    3 h
    1 month
    6 months
    Storage Storage temperature Wet ice
    -20 degrees C
    -80 degrees C
    Analyte Extraction and Purification Protease inhibitor Cocktail
    Biospecimen Aliquots and Components Centrifugation Multiple speeds compared
    View more

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