Research SummaryIntermediate Fellowship research summary
The research has two distinct goals aimed at development of new methods for biomedical applications.
Heat change occurs in most of the biological processes and can be used in bio-sensing. In our laboratory, we develop new types of micro-cantilever based, label-free and electrical read-out sensors that measure the heat change in a typical process such as enzyme catalyzed reactions or ligand-binding. After successful demonstration of these measurements, the sensor will be tested for robust diagnostics applications. This will be further developed into a micro-cantilever based colorimetry to find out the affinity of certain ligands to potential drug targets. The biophysical data obtained will be compared to that of Isothermal Titration Calorimetry (ITC). Since the proposed calorimetry can work with small amounts of proteins it is possible to investigate G-protein coupled receptors (GPCRs) which are difficult to express and purify in large amounts. This will play a significant role in target validation research.
The second goal of the project is to develop a new technique by combining Fluorescence correlation spectroscopy (FCS) and Atomic force Microscopy (AFM). This instrument will facilitate the measurements on protein activity under mechanical stress. AFM is used to apply known strain on single molecules and FCS is used to find out the stability of different confirmations sampled due to thermal fluctuations. As a test of the method developed, we will investigate Ribonuclease A which is a work–horse in protein folding research.