AI Article Synopsis
- Saliva is a promising source for biomarkers of oral and systemic diseases, and a standardized protocol has been developed for analyzing whole saliva using SELDI-TOF/MS.
- The study examined the effects of various factors like storage time, freeze/thaw cycles, and sample treatment on the quality of protein profiles in saliva while noting differences related to sex and time of day.
- The findings indicated that saliva maintains stability for a few hours at room temperature and up to five freeze/thaw cycles, with specific protocols enhancing analysis quality, paving the way for identifying disease-related biomarkers.
Article Abstract
Background: Saliva is one of the most promising and easy-to-collect source of potential biomarkers of oral and systemic disease. We standardized a protocol suitable for pre-analytical treatment and for the analysis of whole normal saliva by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF/MS).
Methods: We evaluated the impact of storage time, freeze/thaw cycles, denaturing agents, glycoproteins depletion, centrifugation, type of matrix and ProteinChip used on the quality of the SELDI protein profile. Moreover, we explored the inter-individual and between-sex differences and the changes in the sample composition over the day.
Results: Saliva was qualitatively stable, in the absence of protease inhibitors, for up to 3 h from the collection at room temperature, although the intensity of a number of peaks slightly decreased between 0 and 3 h and the addition of protease inhibitors did not completely revert this trend. The saliva proteome changed during the day and showed relevant between-sex differences. The protein profile remained stable for up to five freeze/thaw cycles. The addition of denaturing solutions and the depletion of glycoproteins improved the quality of the spectra without affecting their reproducibility.
Conclusions: We defined a protocol that improved the quality and the reproducibility of SELDI-TOF/MS analysis, thus potentially supporting the search for putative biomarkers of disease.
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| http://dx.doi.org/10.1515/CCLM.2008.013 | DOI Listing |
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