UCLA scientists have developed an automated microfluidic platform that uses on-chip hydrogen-deuterium exchange (HDX) coupled with mass spectrometry (MS) to characterize protein structure.
One of the major challenges in the protein therapeutics field is to define the higher order structure and conformational dynamics that dictate the biological activity, stability and safety of a protein. Limiting both drug development and quality control is the lack of a rapid and reliable analytical method that allows real-time monitoring of protein-ligand/target interactions. Hydrogen-deuterium exchange with mass spectrometry (HDX-MS) has been used to interrogate protein dynamics in solution, but current systems suffer from low reproducibility due to lengthy and complicated operations. In particular, analysis of integral membrane proteins is especially difficult as their hydrophobic domains limit solubility in aqueous solvents. Since membrane proteins are the site of action of more than 50% of known drugs, progress in the protein therapeutics field would benefit from a high-throughput method to conveniently analyze the structure and dynamics of proteins in solution in physiologically relevant concentrations, alone and when complexed with various ligands.
UCLA scientists have developed a Digital Microfluidic Droplet Generator (DMDG) chip that functions as an automated platform suitable for use in rapid hydrogen-deuterium exchange (HDX) experiments. This platform is fully compatible with various liquid chromatography and electrospray ionization mass spectrometry (ESI-MS) systems. This innovation can be used to investigate drug-protein interactions, map enzyme binding pockets, and characterize protein folding dynamics. Pharmaceutical and biotechnology companies can utilize this device to streamline their drug discovery and development, and for quality control of their protein-based therapeutics
Researchers have developed a working prototype and have conducted proof-of-concept HDX-MS experiments using DMDG chips.
|Patent Cooperation Treaty||Reference for National Filings||WO2012167273||12/06/2012||2011-556|