Immobilized‐enzyme reactors integrated with capillary electrophoresis for pharmaceutical research

Abstract

Enzymes play an essential role in many aspects of pharmaceutical research as drug targets, drug metabolizers, enzyme drugs and more. In this specific field, enzyme assays are required to meet a number of specific requirements, such as low cost, easy automation, and high reliability. The integration of an immobilized-enzyme reactor to capillary electrophoresis represents a unique approach to fulfilling these criteria by combining the benefits of enzyme immobilization, that is, increased stability and repeated use, as well as the minute sample consumption, short analysis time, and efficient analysis provided by capillary electrophoresis. In this review, we summarize, analyze, and discuss published works where pharmaceutically relevant enzymes were used to prepare capillary electrophoresis-integrated immobilized-enzyme reactors in an online manner. The presented assays are divided into three distinct groups based on the drug-enzyme relationship. The first, more extensively studied group employs enzymes that are considered to be therapeutic targets, the second group of assays present tools to assess drug metabolism and the third group assesses enzyme drugs. Furthermore, we examine various methods of enzyme immobilization and their implications for assay properties.

Controllable and high‐performance immobilized enzyme reactor: DNA‐directed immobilization of multienzyme in polyamidoamine dendrimer‐functionalized capillaries

“…Of these techniques, CE has great potential because it is characterized by rapid analysis, diverse separation modes, high separation efficiency, and low sample consumption, and it is applicable to biomacromolecule assays. Currently, CE‐integrated IMERs are widely used in various fields, such as studies on the kinetics of enzyme catalytic mechanism and enzyme inhibitor screening .…”

“…Many complex reaction systems in nature require the participation of at least two different enzymes [15][16][17][18][19][20][21]. However, most open-tubular CE-integrated IMERs are limited to the immobilization of single enzyme, which significantly confines their broad practical application.…”

Immobilizing sulfotransferase 1A1 on magnetic microparticles and their evaluation using capillary electrophoresis

“…It is a separation technique making limited use of solvents thus ensuring limited wastage. As mentioned in several reviews, an increasing number of groups has been focusing on CE based immobilized micro enzyme reactors and a very recent review has discussed the implementation of CE‐IMERs for pharmaceutical research .…”

“…Enzyme immobilization for the creation of CE-IMERs is mostly done on either the inner capillary wall or magnetic nano/microparticles [8,9].…”

Capillary electrophoresis‐based enzyme assays for β‐lactamase enzymes

“…In the case of β‐lactamases, various formats have been employed including photometric, biochemical, mass spectrometric or polymerase chain reaction‐based and other sequencing assays as summarized, for example, in . Due to the high efficiency and minute sample requirements, CE has been shown to be a powerful microanalytical technique for the determination of the activity of enzymes including the characterization of substrates and inhibitors . With regard to penicillinases, CE–MS has been used to identify β‐lactamase‐derived proteins .…”