Journal of Molecular Biomarkers & Diagnosis

Protein Map Standardization of Human Saliva Using Two Dimensional Gel Electrophoresis (2-DE)

Abstract

Saeid Reza Doustjalali, Ali Yaldrum, Karim Al-Jashamy, Mohammed Irfan, Khin Thant Zin, Nyan Htain Linn, Wai Ma Lin, Vinothini Appalanaidu, Samiah Yasmin Abdul Kadir, Jeyaseelan Nadankutty, Rohaini Mohamad, Wong A-Chin, Htet Htet, Ahmad Yusuf, Rebecca SY Wong, Vinoth Kumarasamy, Christinal PW Teh, Nazrila SF Suhaimi, Hafiza Arzuman, Aida Nur Ashikin A

Over the past decade, the use of saliva as an auxiliary diagnostic tool for biomarker detection has gained considerable acceptance as a non-invasive, inexpensive alternative to conventional serum method. In proteomics, the most promising technique which can be used for detecting salivary biomarker with sufficient resolving power is two-dimensional gel electrophoresis (2-DE) that provides a unique platform for the simultaneous separation of proteins in a complex mixture. However, as a fact most of the new scientists in developing countries are still facing problems with optimization of 2-DE techniques because the performance of optimization techniques in proteomics research using 2-DE has its own limitations. Therefore, the present study was established to generate a reproducible and optimized protocol to display the 2-DE protein map of human saliva. Our results showed the standard optimization of the 2-DE protein mapping was achieved at pH 3-10 with 60 μg proteins loading. This protocol could be used by other scientists along with identification of differentially expressed proteins by mass spectrometry when 2-DE protein map of patients saliva are compared to that of normal healthy individuals. These differentially expressed proteins could be later used as specific and sensitive biomarkers for early diagnosis and prognosis of the disease in question. In conclusion, the procedure used in our study generated a highly reproducible and optimized reference 2-DE protein mapping of human saliva.