An On-Column Enzyme Mediated Fluorescence-Amplification Method for Plasma Total Cholesterol Measurement by Capillary Electrophoresis with LIF Detection

Zhou, Guobin; Guan, Yueqing

doi:10.1007/s10337-016-3023-2

Cited by 12 publications

(7 citation statements)

References 35 publications

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“…Aminophenyl fluorescein was originally purchased from Merck or the Cayman Chemical Company (distributed by Biomol GmbH, Hamburg, Germany) and subsequently synthesised as described in the literature. [38] Detailed protocols for the synthesis and purification of aminophenyl fluorescein are provided in Section S2 of the Supporting Information.…”

Section: Methodsmentioning

confidence: 99%

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

et al. 2020

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Halide assays are important for the study of enzymatic dehalogenation, a topic of great industrial and scientific importance. Here we describe the development of a very sensitive halide assay that can detect less than a picomole of bromide ions, making it very useful for quantifying enzymatic dehalogenation products. Halides are oxidised under mild conditions using the vanadium‐dependent chloroperoxidase from Curvularia inaequalis, forming hypohalous acids that are detected using aminophenyl fluorescein. The assay is up to three orders of magnitude more sensitive than currently available alternatives, with detection limits of 20 nM for bromide and 1 μM for chloride and iodide. We demonstrate that the assay can be used to determine specific activities of dehalogenases and validate this by comparison to a well‐established GC‐MS method. This new assay will facilitate the identification and characterisation of novel dehalogenases and may also be of interest to those studying other halide‐producing enzymes.

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Section: Methodsmentioning

confidence: 99%

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

et al. 2020

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“…In addition, other components of biological fluids could be targets using the EMMA-CE-LIF platform. The same group reported this method in the use of determining total cholesterol in human plasma [111].…”

Section: Ce-lif-based Enzymatic Assaysmentioning

confidence: 99%

Application of Capillary Electrophoresis with Laser-Induced Fluorescence to Immunoassays and Enzyme Assays

Nguyen

Kang

2019

Molecules

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Capillary electrophoresis using laser-induced fluorescence detection (CE-LIF) is one of the most sensitive separation tools among electrical separation methods. The use of CE-LIF in immunoassays and enzyme assays has gained a reputation in recent years for its high detection sensitivity, short analysis time, and accurate quantification. Immunoassays are bioassay platforms that rely on binding reactions between an antigen (analyte) and a specific antibody. Enzyme assays measure enzymatic activity through quantitative analysis of substrates and products by the reaction of enzymes in purified enzyme or cell systems. These two category analyses play an important role in the context of biopharmaceutical analysis, clinical therapy, drug discovery, and diagnosis analysis. This review discusses the expanding portfolio of immune and enzyme assays using CE-LIF and focuses on the advantages and disadvantages of these methods over the ten years of existing technology since 2008.

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“…Then they developed a similar method again to determine the plasma total cholesterol. In this study, the diluted samples were also spiked with APF and the mixed enzyme solutions contained cholesterol oxidase and HRP [54] . All recent studies related to EMMA are summarized in Table 1 .…”

Section: In-capillary Enzyme Assaysmentioning

confidence: 99%

Recent advances in screening of enzymes inhibitors based on capillary electrophoresis

Cheng

Chen

2018

Journal of Pharmaceutical Analysis

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Capillary electrophoresis with many advantages plays an important role in pharmaceutical analysis and drug screening. This review gives an overview on the recent advances in the developments and applications of capillary electrophoresis in the field of enzyme inhibitor screening. The period covers 2013 to 2017. Both the pre-capillary enzyme assays and in-capillary enzyme assays which include electrophoretically mediated microanalysis (EMMA) and immobilized enzyme microreactor (IMER) are summarized in this article.

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An On-Column Enzyme Mediated Fluorescence-Amplification Method for Plasma Total Cholesterol Measurement by Capillary Electrophoresis with LIF Detection

Cited by 12 publications

References 35 publications

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

An Ultrasensitive Fluorescence Assay for the Detection of Halides and Enzymatic Dehalogenation

Application of Capillary Electrophoresis with Laser-Induced Fluorescence to Immunoassays and Enzyme Assays

Recent advances in screening of enzymes inhibitors based on capillary electrophoresis

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