Mn2+-Mediated Modulation of PfAgo Activity for Biosensing

Ruijie Fu; Jinjie Hou; Zexiang Wang; Yunlei Xianyu

doi:10.1002/adhm.202304484

Mn²⁺-Mediated Modulation of PfAgo Activity for Biosensing

Adv Healthc Mater. 2024 Mar 26:e2304484. doi: 10.1002/adhm.202304484. Online ahead of print.

Authors

Ruijie Fu^{1

2}, Jinjie Hou^{1

2}, Zexiang Wang^{1

3}, Yunlei Xianyu^{1

2

4}

Affiliations

¹ College of Biosystems Engineering and Food Science, Zhejiang University, Zhejiang, 310058, China.
² Key Laboratory of Precision Medicine in Diagnosis and Monitoring Research of Zhejiang Province, Sir Run Run Shaw Hospital, Zhejiang, 310058, China.
³ Institute of Pesticide and Environmental Toxicology, Key Laboratory of Biology of Crop Pathogens and Insects of Zhejiang Province, Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Zhejiang, 310058, China.
⁴ Department of Clinical Laboratory, Sir Run Run Shaw Hospital, Zhejiang University School of Medicine, Zhejiang, 310058, China.

PMID: 38530141
DOI: 10.1002/adhm.202304484

Abstract

Argonaute (Ago) as a powerful enzyme has provided new insights into biosensing due to its programmability, high sensitivity, and user-friendly operation. However, current strategies mainly rely on phosphorylated guide DNA to modulate the cleavage activity of Ago, which is limited in versatility and simplicity. Herein, the authors report the Mn²⁺-enhanced cleavage activity of Ago and employ Mn-ions with variable valence to regulate the activity of Pyrococcus furiosus Ago (PfAgo) for biosensing applications. The conversion of Mn ions with different valence states through MnO₂ nanoflowers enables the sensitive detection of ascorbic acid, alkaline phosphatase, and arsenic with limits of detection of 2.5 nmol L^-1, 0.009 U L^-1, and 0.4 ng mL^-1, respectively. A PfAgo-based immunoassay is further developed that allows for the detection of diverse targets, thus providing a promising toolbox to broaden PfAgo-based sensors into versatile bioanalytical and biomedical applications.

Keywords: Mn ion; MnO2 nanoflower; Pyrococcus furiosus argonaute; biosensing; cleavage activity.

Abstract

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