Understanding the role of proteins, peptides, and their post-translational modifications in the metabolism of healthy and diseased tissue is crucial to elucidate biological mechanisms, identify potential drug targets, or define disease biomarkers. Yet biological change begins from the moment a tissue sample is removed from its native environment. This disruption of normal control mechanisms causes drastic alterations at the molecular level. This is manifested by alterations in phosphorylation states, the breakdown of proteins due to uncontrolled proteolytic activity, and changes to metabolite levels.
In short, sample quality deteriorates rapidly, and vital information about the in vivo components may be lost or distorted. With such a reduction in sample quality, the risks of inter-sample variation, incorrect data interpretation, and drawing misleading conclusions increase substantially. These post-mortem biological changes present an analytical challenge and must be prevented to ensure reliable, high-quality data.
Scientists at Denator have developed a heat-stabilization technology that utilizes conductive heating, under controlled pressure, to rapidly and irreversibly denature the proteins in a tissue sample. The complete and permanent elimination of enzymatic activity that would otherwise cause biological change preserves the original composition and state of proteins, peptides, and post-translational modifications. This increases the accuracy and quality of analytical results.
After heat stabilization, of either fresh or frozen tissue samples, in a Stabilizor system, biological components can be extracted and analyzed using common buffers and techniques.
It has been demonstrated that kinase and phosphatase activities, critical in the control of phosphorylation states, are eliminated in heat-stabilized tissue. A comparison with snap-frozen samples showed phosphatase activity in heat-stabilized samples equivalent to background assay levels. Even in the presence of inhibitors, snap-frozen samples contained significantly higher enzyme activity.