Recent advances in <scp>CE</scp>‐mediated microanalysis for enzyme study

Wang, Xu; Li, Kefeng; Adams, Erwin; Schepdael, Ann Van

doi:10.1002/elps.201300294

Cited by 45 publications

(40 citation statements)

References 70 publications

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“…3b-e). The measured potential over time compared to the amount of reactant added can be used to obtain the sensitivity of each thermopile using equation ( (4) in which ΔH (μJ/mole) is the enthalpy of the reaction, which was obtained using isothermal titration calorimetry (ITC). Reaction enthalpies obtained from ITC were preferred over literature values since the enthalpy can change with experimental conditions.…”

Section: Conventional Calibration Methods Are Not Suitable For Calibrmentioning

confidence: 99%

“…Therefore various label-free methods for enzyme activity measurements have been developed, based on different techniques, such as liquid chromatography-mass spectrometry (LC-MS) [3], capillary electrophoresis [4,5], a method based on macrocycle-fluorescent dye complex formation [6], and methods based on heat measurements with isothermal titration calorimetry (ITC) [7]. Recently, a method for timeresolved initial rate measurements using isothermal titration calorimetry (ITC) has been developed as a generic assay for the activity of enzymes.…”

Section: Introductionmentioning

confidence: 99%

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Fast and accurate enzyme activity measurements using a chip-based microfluidic calorimeter

Schie

Ebrahimi

Hagen

et al. 2018

Analytical Biochemistry

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Section: Conventional Calibration Methods Are Not Suitable For Calibrmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Fast and accurate enzyme activity measurements using a chip-based microfluidic calorimeter

Schie

Ebrahimi

Hagen

et al. 2018

Analytical Biochemistry

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“…The most important example in this field is the use of immobilized enzyme biomicroreactors in capillary electrophoresis [123,124], a well-known analytical technique where an electric field is applied to induce the separation of analytes on the basis of their electrophoretic mobility. Bio-microreactors for capillary electrophoresis are discussed separately in this paper because their preparation requires special approaches.…”

Section: Magnetic Beadsmentioning

confidence: 99%

Enzymatic microreactors in biocatalysis: history, features, and future perspectives

Laurenti¹,

Vianna²

2016

Biocatalysis

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Microfluidic reaction devices are a very promising technology for chemical and biochemical processes. In microreactors, the micro dimensions, coupled with a high surface area/volume ratio, permit rapid heat exchange and mass transfer, resulting in higher reaction yields and reaction rates than in conventional reactors. Moreover, the lower energy consumption and easier separation of products permit these systems to have a lower environmental impact compared to macroscale, conventional reactors. Due to these benefits, the use of microreactors is increasing in the biocatalysis field, both by using enzymes in solution and their immobilized counterparts. Following an introduction to the most common applications of microreactors in chemical processes, a broad overview will be given of the latest applications in biocatalytic processes performed in microreactors with free or immobilized enzymes. In particular, attention is given to the nature of the materials used as a support for the enzymes and the strategies employed for their immobilization. Mathematical and engineering aspects concerning fluid dynamics in microreactors were also taken into account as fundamental factors for the optimization of these systems.

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“…Since then, all aspects of enzyme-related analysis have been studied by CE methods including the evaluation of enzymatic activity, enzyme kinetics, identification and characterization of enzyme inhibitors and activators as well as metabolic pathways as, for example, summarized in [5][6][7][8][9][10][11][12].…”

Section: Introductionmentioning

confidence: 99%

“…Analyses employing microfluidic devices were only included when electrophoretic separations were performed. Earlier literature on CE enzyme assays has been summarized, for example, in [5][6][7][8][9][10][11]. The CE-based investigation of the in vitro drug metabolism by cytochrome P450 (CYP) enzymes has also been reviewed [12,13].…”

mentioning

confidence: 99%

Advances in Capillary Electrophoresis-Based Enzyme Assays

Scriba

Belal

2015

Chromatographia

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IntroductionEnzymes catalyze metabolic reactions in cells regulating homeostasis, growth and reproduction of living organisms. Enzymes are also often involved in human disease. The analysis of the human genome has revealed that within the so-called druggable genome a significant portion (in some analyses more than half) of the potential drug targets are enzymes [1][2][3]. Consequently, enzymes are important targets in the development of new drugs. Moreover, enzymes play a role in the clinical diagnosis of diseases. Therefore, effective methods for characterization of enzymes as well as for the determination of their activity are important, for example, for the understanding of enzyme-mediated metabolic reactions or the identification of substrates and inhibitors for the treatment of human diseases.Capillary electrophoresis (CE) has been applied to enzyme analysis for more than 20 years since the first report by Banke et al. on an assay of alkaline protease [4]. Since then, all aspects of enzyme-related analysis have been studied by CE methods including the evaluation of enzymatic activity, enzyme kinetics, identification and characterization of enzyme inhibitors and activators as well as metabolic pathways as, for example, summarized in [5][6][7][8][9][10][11][12].Generally, CE-based enzyme assays can be divided in three groups, pre-capillary (offline) assays, in-capillary (online) assays and post-capillary assays. In the pre-capillary assays, all required components including enzyme, substrate, co-factors, etc., are mixed in a vial and incubated for an intended period of time. Subsequently, the reaction is quenched and an aliquot is subjected to CE analysis for the determination of substrate(s) and/or co-substrate(s) and/ or product(s). Multiple sampling or repeated sampling in combination with sequential runs for the determination of Abstract Capillary electrophoresis (CE) has become a flexible and accurate, high-efficiency analytical separation technique in many areas requiring only minute amounts of sample and chemicals. Thus, CE has also been recognized as a suitable technique to study enzymatic reactions including the determination of Michaelis-Menten kinetic data or the identification and characterization of inhibitors. The most often applied CE-based enzyme assay modes can be divided into two categories: (1) pre-capillary assays where incubations are performed offline followed by CE analysis of substrate(s) and/or product(s) and (2) in-capillary assays in which the enzymatic reaction and analyte separation are performed in the same capillary. In case of the in-capillary assays, the enzyme may be immobilized or in solution. The latter is also referred to as electrophoretically mediated microanalysis (EMMA), while in the case of immobilized enzyme the term immobilized enzyme reactor (IMER) is used. The present review summarizes the literature on CEbased enzyme assays published between January 2010 and April 2015. Immobilized enzyme reactors as well as microfluidic devices applied to the study of enzymatic a...

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Recent advances in CE‐mediated microanalysis for enzyme study

Cited by 45 publications

References 70 publications

Fast and accurate enzyme activity measurements using a chip-based microfluidic calorimeter

Fast and accurate enzyme activity measurements using a chip-based microfluidic calorimeter

Enzymatic microreactors in biocatalysis: history, features, and future perspectives

Advances in Capillary Electrophoresis-Based Enzyme Assays

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