Standardization of a protocol for shotgun proteomic analysis of saliva

Authors

  • Talita Mendes da Silva Ventura Universidade de São Paulo, Faculdade de Odontologia de Bauru, Departamento de Ciências Biológicas, Bauru, São Paulo http://orcid.org/0000-0003-2101-1350
  • Nathalia Regina Ribeiro Universidade de São Paulo, Faculdade de Odontologia de Bauru, Departamento de Ciências Biológicas, Bauru, São Paulo http://orcid.org/0000-0002-6086-6303
  • Aline Salgado Dionizio Universidade de São Paulo, Faculdade de Odontologia de Bauru, Departamento de Ciências Biológicas, Bauru, São Paulo http://orcid.org/0000-0002-8687-0124
  • Isabela Tomazini Sabino Universidade de São Paulo, Faculdade de Odontologia de Bauru, Departamento de Ciências Biológicas, Bauru, São Paulo
  • Marília Afonso Rabelo Buzalaf Universidade de São Paulo, Faculdade de Odontologia de Bauru, Departamento de Ciências Biológicas, Bauru, São Paulo http://orcid.org/0000-0002-5985-3951

DOI:

https://doi.org/10.1590/1678-7757-2017-0561

Keywords:

Methods, Proteomics, Standartization, Saliva

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.

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Published

2022-09-20

Issue

Section

Original Articles

How to Cite

Ventura, T. M. da S., Ribeiro, N. R., Dionizio, A. S., Sabino, I. T., & Buzalaf, M. A. R. (2022). Standardization of a protocol for shotgun proteomic analysis of saliva. Journal of Applied Oral Science, 26, e20170561. https://doi.org/10.1590/1678-7757-2017-0561