A simple, real-time assay of horseradish peroxidase using biolayer interferometry

Author:

Kojima Takaaki1,Nakane Ayako1,Zhu Bo1,Alfi Almasul1,Nakano Hideo1

Affiliation:

1. Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya, Japan

Abstract

ABSTRACT Horseradish 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.

Funder

Ministry of Education, Culture, Sports, Science, and Technology of Japan

Grant-in-Aid for Young Scientists

Publisher

Oxford University Press (OUP)

Subject

Organic Chemistry,Molecular Biology,Applied Microbiology and Biotechnology,General Medicine,Biochemistry,Analytical Chemistry,Biotechnology

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