Enzyme inhibitor screening by capillary electrophoresis with an on-column immobilized enzyme microreactor created by an ionic binding technique

Analytical Chemistry
Zhong-Mei Tang, Jing-Wu Kang

Abstract

A novel strategy for screening the enzyme inhibitors from the complex mixtures by capillary electrophoresis with an on-column immobilized enzyme microreactor created by an ionic binding technique is reported. The enzyme microreactor was prepared in two steps: First, the capillary wall was dynamically coated with a polycationic electrolyte hexadimethrine bromide (HDB) by simply flushing the column using the HDB solution. Subsequently, a plug of the enzyme solution was injected and incubated for 5 min to permit the enzyme molecules to immobilize on the positively charged coating via ionic binding. To demonstrate this strategy, angiotensin-converting enzyme (ACE) was employed as a model for the enzyme immobilization, inhibition study, and inhibitor screening. It has been proved that such a prepared immobilized ACE microreactor displays a high enough activity and stability. Furthermore, the immobilized enzyme microreactor could be easily renewed. The inhibition study or inhibitor screening was accomplished through the following procedure: (i) the substrate solution was injected and incubated within the microreactor for a short time span; (ii) subsequently, the voltage was applied to separate the product of the enzyme reaction from ...Continue Reading

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Citations

Apr 9, 2013·Analytica Chimica Acta·Meera Shanmuganathan, Philip Britz-McKibbin
May 20, 2008·Analytical Chemistry·Vratislav KostalEdgar A Arriaga
Jun 29, 2013·Journal of Pharmaceutical and Biomedical Analysis·Ting-Fu JiangZhi-Hua Lv
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Oct 11, 2017·Journal of Separation Science·Jan Schejbal, Zdeněk Glatz
Apr 27, 2021·Colloids and Surfaces. B, Biointerfaces·Sizhu RenYanlin Yuan

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