A simple,real-time assay of horseradish peroxidase using biolayer interferometry |
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Authors: | Takaaki Kojima Ayako Nakane Bo Zhu Almasul Alfi |
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Institution: | 1. Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya, Japankojimat@agr.nagoya-u.ac.jphttps://orcid.org/0000-0001-7282-121X;3. Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya, Japan |
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Abstract: | ABSTRACTHorseradish peroxidase (HRP) isoenzyme C1a is one of the most widely used enzymes for various analytical methods in bioscience research and medical fields. In these fields, real-time monitoring of HRP activity is highly desirable because the utility of HRP as a reporter enzyme would be expanded. In this study, we developed a simple assay system enabling real-time monitoring of HRP activity by using biolayer interferometry (BLI). The HRP activity was quantitatively detected on a BLI sensor chip by tracing a binding response of tyramide, a substrate of HRP, onto an immobilized protein. This system could be applied to analyses related to oxidase activity, as well as to the functional analysis of recombinant HRP. |
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Keywords: | Horseradish peroxidase (HRP) heterologous expression biolayer interferometry (BLI) real-time enzymatic assay |
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