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Electrowetting-based Digital Microfluidics Platform for Automated Enzyme-linked Immunosorbent Assay
08:22

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Published on: February 23, 2020

Capillary electrophoretic enzyme assays.

Gerhard K E Scriba1, Hans Abromeit, Martina Hense

  • 1Department of Pharmaceutical Chemistry, School of Pharmacy, Friedrich Schiller University, Jena, Germany. gerhard.scriba@uni-jena.de

Methods in Molecular Biology (Clifton, N.J.)
|February 7, 2013
PubMed
Summary
This summary is machine-generated.

Capillary electrophoresis (CE) offers efficient enzyme assays with minimal sample and chemical use. Both offline (pre-capillary) and online (in-capillary) CE methods are detailed, including electrophoretically mediated microanalysis (EMMA).

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Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method
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Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method
07:58

Highly Sensitive and Quantitative Detection of Proteins and Their Isoforms by Capillary Isoelectric Focusing Method

Published on: September 19, 2018

Area of Science:

  • Analytical Chemistry
  • Biochemistry

Background:

  • Capillary electrophoresis (CE) is increasingly utilized for enzyme assays due to its high efficiency, versatility, small sample requirements, and low chemical consumption.
  • CE enzyme assays are broadly categorized into pre-capillary (offline) and in-capillary (online) approaches.

Purpose of the Study:

  • To describe and exemplify both pre-capillary and in-capillary CE techniques for enzyme assays.
  • To illustrate the application of these methods using specific enzymatic reactions.

Main Methods:

  • Pre-capillary assays involve offline incubation followed by CE analysis of substrates and products.
  • In-capillary assays integrate enzyme reaction and analyte separation within the same capillary, using either immobilized enzymes or solution-phase enzymes.
  • Electrophoretically mediated microanalysis (EMMA) is highlighted as an online technique where incubation and analysis rely on electrophoretic phenomena.

Main Results:

  • The chapter details the application of CE for enzyme assays, demonstrating its effectiveness in analyzing enzymatic reactions.
  • Specific examples include the deacetylation of acetyl-lysine residues by sirtuin enzymes and the hydrolysis of acetylthiocholine by acetylcholinesterase.

Conclusions:

  • Both pre-capillary and in-capillary CE methods provide robust platforms for enzyme activity analysis.
  • CE, particularly EMMA, offers a powerful and efficient approach for studying enzyme kinetics and substrate-product relationships.