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(Reference retrieved automatically from SciELO through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Standardization of a protocol for shotgun proteomic analysis of saliva

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Author(s):
Talita Mendes da Silva VENTURA ; Nathalia Regina RIBEIRO ; Aline Salgado DIONIZIO ; Isabela Tomazini SABINO ; Marília Afonso Rabelo BUZALAF
Total Authors: 5
Document type: Journal article
Source: Journal of Applied Oral Science; v. 26, p. -, 2018.
Web of Science Citations: 0
Abstract

Abstract Saliva contains numerous proteins and peptides, each of them carries a number of biological functions that are very important in maintaining the oral cavity health and also yields information about both local and systemic diseases. Currently, proteomic analysis is the basis for large-scale identification of these proteins and discovery of new biomarkers for distinct diseases. Objective This study compared methodologies to extract salivary proteins for proteomic analysis. Material and Methods Saliva samples were collected from 10 healthy volunteers. In the first test, the necessity for using an albumin and IgG depletion column was evaluated, employing pooled samples from the 10 volunteers. In the second test, the analysis of the pooled samples was compared with individual analysis of one sample. Salivary proteins were extracted and processed for analysis by LC-ESI-MS/MS. Results In the first test, we identified only 35 proteins using the albumin and IgG depletion column, while we identified 248 proteins without using the column. In the second test, the pooled sample identified 212 proteins, such as carbonic anhydrase 6, cystatin isoforms, histatins 1 and 3, lysozyme C, mucin 7, protein S100A8 and S100A9, and statherin, while individual analysis identified 239 proteins, among which are carbonic anhydrase 6, cystatin isoforms, histatin 1 and 3, lactotransferrin, lyzozyme C, mucin 7, protein S100A8 and S100A9, serotransferrin, and statherin. Conclusions The standardization of protocol for salivary proteomic analysis was satisfactory, since the identification detected typical salivary proteins, among others. The results indicate that using the column for depletion of albumin and IgG is not necessary and that performing individual analysis of saliva samples is possible. (AU)

FAPESP's process: 17/05031-2 - Proteomic analysis of the acquired enamel pellicle and saliva in patients with head and neck cancer that are submitted radiotherapy
Grantee:Talita Mendes Oliveira Ventura
Support type: Scholarships in Brazil - Doctorate