The discovery of proteinaceous disease biomarkers and their clinical validation is critically important for the enablement of molecular diagnostics and ultimately, precision medicine. In spite of the importance of biomarkers, research done in the last two decades has yielded limited success in enhancing the repertoire of protein biomarkers of disease. This apparent dichotomy is in large part due to the fact that the process from discovery to validation is complex and involves a multidisciplinary collaboration across clinical, biochemical and biophysical disciplines and that that many initiatives sought to identify unique markers for a given disease with perfect correlation. Recent advances in better biomarker discovery strategies, clinical sampling and sample storage, better sample preparation methods that consider the inherent instabilities associated with clinical specimens, better discovery methods and analytical instrumentation suited for assay validation and clinical assays together promise to generate better panel tests for disease biomarkers of clinical significance.