151. In situ monitored vortex fluidic-mediated protein refolding/unfolding using an aggregation-induced emission bioprobe
- Author
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Xinyi Zhang, Haozhen Hu, Qi Hu, Kyle Fan, Yuning Hong, Youhong Tang, and Colin L. Raston
- Subjects
Protein Denaturation ,Protein Folding ,Circular dichroism ,Protein Conformation ,Microfluidics ,Pharmaceutical Science ,Organic chemistry ,Biosensing Techniques ,Lactoglobulins ,02 engineering and technology ,010402 general chemistry ,01 natural sciences ,Protein Refolding ,Article ,Analytical Chemistry ,chemistry.chemical_compound ,vortex fluidic device ,QD241-441 ,Drug Discovery ,Humans ,Denaturation (biochemistry) ,Cysteine ,Physical and Theoretical Chemistry ,Guanidine ,Protein Unfolding ,Uncategorized ,Chemistry ,Circular Dichroism ,Proteins ,protein folding/unfolding ,021001 nanoscience & nanotechnology ,0104 chemical sciences ,aggregation induced emission ,Kinetics ,Spectrometry, Fluorescence ,Proteostasis ,Chemistry (miscellaneous) ,Unfolded protein response ,Biophysics ,Molecular Medicine ,Protein folding ,0210 nano-technology - Abstract
Protein folding is important for protein homeostasis/proteostasis in the human body. We have established the ability to manipulate protein unfolding/refolding for β-lactoglobulin using the induced mechanical energy in the thin film microfluidic vortex fluidic device (VFD) with monitoring as such using an aggregation-induced emission luminogen (AIEgen), TPE-MI. When denaturant (guanidine hydrochloride) is present with β-lactoglobulin, the VFD accelerates the denaturation reaction in a controlled way. Conversely, rapid renaturation of the unfolded protein occurs in the VFD in the absence of the denaturant. The novel TPE-MI reacts with exposed cysteine thiol when the protein unfolds, as established with an increase in fluorescence intensity. TPE-MI provides an easy and accurate way to monitor the protein folding, with comparable results established using conventional circular dichroism. The controlled VFD-mediated protein folding coupled with in situ bioprobe AIEgen monitoring is a viable methodology for studying the denaturing of proteins.
- Published
- 2021
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