Development of a high-throughput, label-free optical system to evaluate the interaction of enzymes involved in inflammation with cells
Various proteins, including the members of blood plasma enzyme cascade, act on different cells (such as endothelial cells) via multiple signaling pathways. The detailed comprehension of the interaction between these proteins and cells is substantially important for the efficient diagnosis, prognosis and therapy of inflammatory vascular diseases (e.g. sepsis or hereditary angioedema). The real-time and label-free detection of the complex function of cells cannot be easily accomplished in high-throughput-screening system. József Dobó has been studying the blood plasma serine proteases. He has experience with the preparation of these enzymes (both in recombinant and plasma-derived), and the analysis of commercial enzymes. Therefore, he has the capability to provide the other cooperation partners with enzymes that cannot be purchased (MASP-1, MASP-2, MASP-3) or to further purify commercial enzymes. He also has the infrastructure and know-how to synthesize specific enzyme inhibitors. László Cervenak is involved in endothelial cell studies to clarify the endothelial cell component of the inflammatory reaction. He has the ability to adapt different cell type to various cell culture surfaces, and he can validate these attachments with several cellular test systems. Róbert Horváth is studying the setup and optimization of different nanobiosensor systems, and utilizes these methods to protein- and cellular measurements. He has the experience in the development and biological application of dedicate nanobiosensor instruments (some of them are in prototype phase). The methods and instruments he and his group developed are completely applicable to the high-throughput screening of complex, real-time cellular responses by label-free systems.
The cooperation of the members aims to set up such a label-free, real-time measurement system for the delineation of inflammatory response of endothelial cells. This would provide an ideal screening platform for blood plasma enzyme system, since their role in endothelial cell activation is recognized to be more- and more important, and related to severe diseases.
László Cervenák - József Dobó - Róbert Horváth