Researchers can study both the disease symptoms and the particular biological processes that lead to their onset and development with modern biophysics. It is through these studies that researchers can identify potential targets for newly developed pharmaceutical drugs. This process, known as drug discovery, often begins with differential scanning fluorimetry or DSF.
What Is DSF?
The origins of many human diseases can be traced back to issues with the structure and function of proteins. For this reason, treatments for disease often involve searching for proteins that can be bound to and targeted by pharmaceutical compounds that can prevent misfolding and other disease-inducing conditions. DSF provides researchers with an advanced, fast-acting tool to examine these proteins.
To begin the process, researchers add fluorescent dye to a solution containing proteins. Then, DSF systems slowly heat the wells of protein samples and monitor the change in fluorescence emitted as the proteins unfold. These changes reveal critical information about the protein’s stability, denaturation process, ligand binding ability, and more. Differential scanning fluorimetry can be used as a high-throughput screening method for libraries of hundreds of proteins. Using high-throughput screening, researchers can cut the time needed to identify a potential drug target from days to mere hours.
Applications of DSF
DSF has multiple applications during the drug discovery process. Applications include:
- Ligand screening. DSF can be used to assess the interaction between potential target proteins’ binding receptors and libraries of ligand compounds that may bind with them. DSF measures the physical and chemical changes of bonds that form by analyzing differences in fluorescence. As a result, researchers can determine which ligands act effectively upon a target protein and binding affinity (the strength of the bond interaction).
- Fragment-binding and linking strategy research. After researchers discover potential “hit” compounds that may target specific disease agents, it is important to determine which compounds interact most effectively with the target in question. Sometimes, small ligand fragments have been found to interact more effectively with the target in question than previously identified compounds. DSF can screen thousands of these fragments, identifying new potential disease inhibitors that may have been missed.
- Protein stability and crystallization optimization. Before target proteins can undergo extensive research, they must undergo purification to ensure any results produced are related to the target protein. Unfortunately, many proteins destabilize and denature – or undergo unfolding and aggregation – during purification or in storage. DSF enables researchers to study the stability of the protein to ensure accurate results.
- Target validation. While effective protein-ligand interactions may occur during research, the same conditions are not present within living cells. As a result, researchers have developed methods of utilizing DSF to perform thermal shift assays on cells, lysates, and tissues. Such studies heat and monitor the target proteins in the cell and assess binding capabilities to validate the target protein for druggable properties.
Through these applications, DSF enables researchers to accomplish the early stages of drug discovery more thoroughly, more effectively, and more quickly than ever before.
Resources:
https://link.springer.com/article/10.1007/s12551-020-00619-2
https://cmi.hms.harvard.edu/differential-scanning-fluorimetry