Quantification of oral cancer biomarkers by targeted proteomic approach

Abstract

The pipeline for candidate biomarker discovery consists of several steps before they are able in clinical practice to safely assist in the diagnosis or prognosis concerning a disease state. Amongst these steps is the verification process, of which the main purpose is to quantify with accuracy and precision the concentration of target proteins/peptides present in the sample of interest. However, many of the candidates do not move forward in the development process of discovery due to the lack of validated quantitative analytical methods in the verification step. The mass spectrometry and its development contribute greatly to the progress of targeted based quantitative proteomic. This technique enables accurate and precise quantitation of biomarkers in a large scale using isotopologues internal standards. Usually, the tandem mass spectrometry is used in the targeted proteomic approach in Selected Reaction Monitoring mode (SRM). Unfortunately, despite the usual selectivity of SRM experiment, the unit resolution of the quadrupole still has drawbacks in quantitative and qualitative determination of biomolecules in complex matrices. Hence, one realizes the importance of high resolution mass spectrometry, which is able to overcome this obstacle. Thus, this project aims to develop and validate an analytical methodology, using hybrid mass spectrometry (linear ion trap /Orbitrap) and isotopic dilution as internal standardization method for quantifying oral cancer biomarkers in complex matrices. This project is expected to be helpful in the development and validation of potential biomarkers and to contribute to its pathway towards clinical trials.