High temporal resolution monitoring of enzyme reaction and inhibition using optically gated vacancy capillary electrophoresis and immobilized enzyme.

作者: Li Yang , Chunyang Chen , Yuanfang Chen , Jing Shi , Sidong Liu

DOI: 10.1016/J.ACA.2010.10.013

关键词:

摘要: A novel method for monitoring of enzyme reaction and inhibition with high temporal resolution was developed by using optically gated vacancy capillary electrophoresis (OGVCE) laser-induced fluorescence (LIF) detection immobilized enzyme. Trypsin cleavage were investigated the presented OGVCE-LIF assay, carboxyfluorescein (FAM) end-labeled Angiotensin as substrate commercially available trypsin. The product continuously loaded into electroosmotic flow while remained in sample vial. Substrate consumption formation monitored simultaneously at 5 s interval during whole time. enzymatic rates obtained from highly consistent. activity Michaelis constants trypsin reaction, well constant (for reversible competitive inhibitor) fraction irreversible inhibitor), obtained. It showed that reported can perform fast, accurate, sensitive reproducible CE assay resolution, thus has great potential application enzyme-substrate systems fast rate fluorescent products.

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