Informace o publikaci
Droplet microfluidics as a high-throughput approach for enzyme kinetics studies
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| Rok publikování | 2023 |
| Druh | Konferenční abstrakty |
| Citace | |
| Popis | Microfluidic systems are recently undergoing rapid development as a high-throughput method useful in bioanalytical applications. Droplet microfluidics is emerging as a promising method in this field. Basic concept of droplet microfluidics is generation of large amount of water based droplets in immiscible oil phase. Since the droplets are isolated and protected by the oil and can be individually manipulated, they can be regarded as separate microreactors. Compared to the conventional technologies, microfluidics offers several advantages such as reduced reagent consumption, shorter analyses times, and rapid production of data in given time. The research of enzyme kinetics is of high interest as many enzymes play their role in human diseases and are potential therapeutical drug targets. The aim is to develop convenient droplet microfluidics based methods for performing enzyme assays with the mentioned advantages. The "hearth" of the device is typically polydimethylsiloxane (PDMS) chip, serving as droplet generator and incubation channel, connected to precise syringe pumps. The enzyme reaction can be monitored using synthetic fluorogenic substrates, offering high sensitivity. For this purpose the microscope is mounted with excitation light source, set of optical filters and photomultiplier tube serving as a detector. The setup allows to measure the reaction rate simply by focusing on different distances in the incubation channel, corresponding to different reaction times. First optimizations of uniform-sized droplets generation as well as pilot study with E. coli beta-galactosidase with fluorogenic substrate as a model system has been done. In the following three years of the doctoral studies the main efforts will be focused on further miniaturization, optimization of the method, and its application for clinically and pharmacologically interesting enzymes. |
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