Microfluidic reaction design for real time chemical reactions monitoring
| Autor: | Abigail H. Casey, Gregory Triplett |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Aqueous solution
Materials science Polydimethylsiloxane Kinetics Microfluidics technology industry and agriculture macromolecular substances Chemical reaction Transduction (biophysics) chemistry.chemical_compound symbols.namesake chemistry Chemical engineering symbols Absorption (chemistry) Raman spectroscopy |
| Zdroj: | Frontiers in Biological Detection: From Nanosensors to Systems XIII. |
| DOI: | 10.1117/12.2575995 |
| Popis: | Microfluidic reactors have become increasingly popular for the characterization of reaction parameters. They have found an important role in the synthesis and monitoring of an expansive list of bioreactions. To study protein phosphorylation, a biochemical reaction that is responsible for a broad range of signal transduction roles, we designed microfluidic reactors based on tubular flow for studying in real time reactor kinetic parameters in protein phosphorylation. In order to monitor phosphorylation as it occurs, we probed the environment where the reaction takes place without influencing the reactor kinetics, while collecting molecular information such as structure and conformation of native proteins. We designed and tested polydimethylsiloxane (PDMS) microfluidic reactors compatible with our confocal Raman spectrometer and overcame significant absorption of important Raman bands from the PDMS. We studied the PDMS device interfaces, determined experimentally which interfaces were least absorbing, and redesigned the microfluidic device using 3D rapid prototyping. Using redesigned devices, we measured Raman spectra on aqueous Adenosine triphosphate (ATP) solutions at varying molar concentrations (0.1 M – 1.0 M) at room temperature over the wavenumber range of 50 - 4000 /cm and accurately observed the behavior of phosphate bonds involved in the protein phosphorylation reaction. We also studied the effects of fluid flow in order to account for effects under transient conditions. To do this, we preloaded the reactor and initiated ATP fluid flow and collected spectra. Our modified design proved to overcome significant signal absorption from PDMS and accurately measured concentrations. |
| Databáze: | OpenAIRE |
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