The rapid assessment of biomolecular kinetics is critical when investigating biological molecular reactions. Contemporary techniques for biomolecular kinetics, such as temperature-jump/stopped-flow and laser-induced temperature-jump platforms, provide some control over temperature and mixing rate, however they are complex in construction and operation, provide limited data and are unable to operate in high-throughput over wide temperature ranges. To address these issues, researchers in the deMello and Arosio groups at ETH and collaborators at Masaryk University have developed a simple yet powerful droplet-based temperature-jump platform for rapidly assessing biomolecular kinetics.
Their droplet-based microfluidic device is able to generate droplets of controlled size and payload, mix droplet contents on ultra-short timescales and then probe reaction kinetics at temperatures between 25 and 80°C. To showcase the capabilities of the platform, the team investigated the elongation kinetics of fibrils of the amyloid β peptide, Aβ 1−42, which is implicated in Alzheimer’s disease. Significantly, the platform is able to monitor fibril growth over a 16 second period, with results being in excellent agreement with bulk elongation data. Additionally, the authors studied the unfolding of the haloalkane dehalogenase, LinB, induced by temperature.
The platform is able to acquire time-resolved spectra over a wide range of temperatures, and thus has broad utility in studying both enzyme kinetics and protein stability at elevated temperatures.
Written by Nan Zong
Read the published article here.