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Issue 18, 2016
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Enzyme catalysis–electrophoresis titration for multiplex enzymatic assay via moving reaction boundary chip

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Abstract

In this work, we developed the concept of enzyme catalysis–electrophoresis titration (EC-ET) under ideal conditions, the theory of EC-ET for multiplex enzymatic assay (MEA), and a related method based on a moving reaction boundary (MRB) chip with a collateral channel and cell phone imaging. As a proof of principle, the model enzymes horseradish peroxidase (HRP), laccase and myeloperoxidase (MPO) were chosen for the tests of the EC-ET model. The experiments revealed that the EC-ET model could be achieved via coupling EC with ET within a MRB chip; particularly the MEA analyses of catalysis rate, maximum rate, activity, Km and Kcat could be conducted via a single run of the EC-ET chip, systemically demonstrating the validity of the EC-ET theory. Moreover, the developed method had these merits: (i) two orders of magnitude higher sensitivity than a fluorescence microplate reader, (ii) simplicity and low cost, and (iii) fairly rapid (30 min incubation, 20 s imaging) analysis, fair stability (<5.0% RSD) and accuracy, thus validating the EC-ET method. Finally, the developed EC-ET method was used for the clinical assay of MPO activity in blood samples; the values of MPO activity detected via the EC-ET chip were in agreement with those obtained by a traditional fluorescence microplate reader, indicating the applicability of the EC-ET method. The work opens a window for the development of enzymatic research, enzyme assay, immunoassay, and point-of-care testing as well as titration, one of the oldest methods of analysis, based on a simple chip.

Graphical abstract: Enzyme catalysis–electrophoresis titration for multiplex enzymatic assay via moving reaction boundary chip

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Supplementary files

Article information


Submitted
14 Jun 2016
Accepted
07 Jul 2016
First published
08 Jul 2016

Lab Chip, 2016,16, 3538-3547
Article type
Paper

Enzyme catalysis–electrophoresis titration for multiplex enzymatic assay via moving reaction boundary chip

R. Zhong, H. Xie, F. Kong, Q. Zhang, S. Jahan, H. Xiao, L. Fan and C. Cao, Lab Chip, 2016, 16, 3538
DOI: 10.1039/C6LC00757K

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