Discovery-based quantitative proteomics identified dozens of novel protein candidates in saliva, yet none are clinically qualified. Mass spectrometry (MS) has been recognized as newer 'gold standard' for the identification and analysis of individual proteins in complex biological samples due to higher sensitivity, specificity and independence of antibody availability. However, whether MS can replace immunoassays for salivary biomarker validation has not been thoroughly studied. Therefore, we aimed to test the potential of label-free quantitative (LFQ) proteomics in comparison to conventional antibody-based methods to quantify ‘candidate biomarkers’ in saliva during experimentally-induced oral inflammation development in humans. Whole unstimulated saliva, collected at five time points, was analyzed by LFQ shotgun proteomics. Salivary levels of ‘candidate biomarkers’ were also assessed by antibody-based immunoassays such as ELISA and time-resolved immunofluorometric assay (IFMA).