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
Conclusion of Paper
Studies
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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:
Analyte Technology Platform Protein SELDI-TOF MS Pre-analytical Factors:
Classification Pre-analytical Factor Value(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