The story behind heat stabilization

Understanding the role of proteins and peptides in normal and diseased tissue is crucial to defining their potential use as drugs, drug targets or disease biomarkers when developing new therapeutics. However, biological changes begin from the moment that a tissue sample is removed from its native environment. This traumatic event can have drastic effects at the molecular level within a very short timeframe. Changes range from alterations of phosphorylation states to general protein cleavage through the activity of enzymes such as proteases, phosphatases and kinases.

Schematic flowchart of the peptidomics experimental setup

In 2003, Karl Sköld and Marcus Söderquist were conducting their graduate studies in the Laboratory for Biological and Medical Mass Spectrometry headed by Per Andrén, at Uppsala University, Sweden. Their main interest was to identify bioactive endogenous neuropeptides in mouse models of Parkinson’s disease, using liquid chromatography and mass spectrometry-based analytical methods. This would enable them to simultaneously follow hundreds of peptides during the course of disease development. At this point in time such peptide profiling was usually carried out with antibody-based methods on one or a few peptides at a time.

The scientists Marcus Söderquist and Karl Sköld in the Laboratory for Biological and Medical Mass Spectrometry, Uppsala University

Success is not always immediate. Like many other researchers around the world, they recognized that post-mortem changes could be distorting their analysis. Despite their efforts to prevent such changes by using traditional methods such as rapid freezing or chemical inhibitors, they still faced the problem of not being able to identify the neuropeptides that should have been present in their tissue samples.

So they reasoned that rapid heating, instead of snap freezing, could permanently denature the enzymes causing these changes thus eliminating potential artifacts. With this revolutionary idea, they joined forces with Karl’s brother Olof in 2005, an established entrepreneur. Together they managed to turn the discovery into a new biotech company, which they named Denator after the method of heat denaturation.

Breakthrough technology

In August 2008, Denator pre-launched the Stabilizor system at HUPO in Amsterdam, one of the largest annual conferences within the field of proteomics. The Stabilizor system utilizes conductive heat, at controlled pressure, to generate rapid, homogenous denaturation of proteins. Denator’s heat stabilization technology now enabled researchers to reveal protein profiles close to in vivo state and identify novel biological molecules that were previously lost due to rapid post mortem changes. Such information is of particular significance to areas such as neuroscience and proteomics research, where short-lived molecules and potential biomarkers need to be confidently identified to understand disease mechanisms and identify potential drug targets or disease markers.