A novel assay for the determination of PreScission protease by capillary electrophoresis

Biophys Chem. 2021 Nov 13;281:106696. doi: 10.1016/j.bpc.2021.106696. Online ahead of print.ABSTRACTThe detection of protease activity in the body plays a significant role in the early diagnosis of diseases. However, enzymes inevitably come into contact with various complex biological fluids in the body during the flow, which greatly increases the detection difficulty. Therefore, protease detection in vivo has great challenges. Herein, we report a new assay for detecting protease using capillary electrophoresis inside a capillary with semicircular bends. We first designed a peptide substrate, and then the peptide was self-assembled with quantum dots to form a QDs-peptide substrate. The capillary was bent to semicircular-shaped turns and served as a micro-reactor to allow protease and substrate react in it. Due to the different electrophoretic velocity, the protease and the substrate were mixed inside the bent capillary with sequential injections and the cleavage of the substrate can be detected using capillary electrophoresis combined with Förster resonance energy transfer technology. This novel assay will greatly expand the detection of enzyme activity in vivo.PMID:34954553 | DOI:10.1016/j.bpc.2021.106696

Source: Biophysical Chemistry - December 26, 2021 Category: Chemistry Authors: Shuwen Zhou Pengfei Cui Jingyu Sheng Xueli Zhang Pengju Jiang Xinye Ni Kai Cao Lin Qiu Source Type: research