Clinical application for the preservation of phospho-proteins through in-situ tissue stabilization

In this report, we re-create laboratory and clinical scenarios for sample collection and test the utility of a new tissue stabilization technique in preserving proteins and protein modifications. In the laboratory setting, tissue stabilization with the Denator Stabilizor T1 resulted in a significantly higher yield of phospho-protein when compared to standard snap freeze preservation. Furthermore, in a clinical scenario, tissue stabilization at collection resulted in a higher yield of total phospho-protein, total phospho-tyrosine, pErkT202/Y204 and pAktS473 when compared to standard methods. Tissue stabilization did not have a significant effect on other post-translational modifications such as acetylation and glycosylation, which are more stable ex-vivo. Tissue stabilization did decrease total RNA quantity and quality.Stabilization at the time of collection offers the potential to better preserve tissue protein and protein modification levels, as well as reduce the variability related to tissue processing delays that are often associated with clinical samples.Recent advances in proteomic technologies have spurred a number of reports examining distinct alterations in protein expression [1,2] or modification [3-6] that are associated with, or can classify, disease states in human patients. Although these biomarker studies provide important analytical and diagnostic tools, a challenge for translational research is the transition of findings from the controlled laboratory environment to the clinical setting, where variation in tissue acquisition and handling practices can introduce significant data variability. This variation can confound data analysis and interpretation, and in turn, impact patient diagnosis and prognosis [7]. Combined with clinical heterogeneity resulting from genetic, physiological, and environmental factors, which are typically controlled for in animal models implemented in the laboratory setting, technical variance introduced during tissue collecti