Issue 15, 2023

Simultaneous quantification of exosomal MMP14 expression and proteolysis activity on a spherical dual-probe-based fluorescent nanosensor

Abstract

Among various exosomal proteins, matrix metalloproteinases (MMPs) are a family of membrane associated endopeptidases and have been considered as potential biomarkers in liquid biopsy owing to their multiple roles in pathological processes. However, the potential of MMP14 expression (MMP14-E) and MMP14 proteolytic activity (MMP14-A) in clinical diagnosis is still not clear due to the lack of sensitive and simultaneous detection techniques. Herein, we propose a fluorescent nanosensor for the simultaneous detection of MMP14-E and MMP14-A using a spherical aptamer/peptide dual-probe strategy. The aptamer and peptide probes were sequentially immobilized on Fe3O4 magnetic nanoparticles coated with gold nanoparticles (m-AuNPs) using a disulfide linker. MMP14 can be specifically recognized by the aptamer, while the proteolytic-active MMP14 can cleave the peptide probe. While achieving simultaneous detection, the proposed sensor provides better analytical performances than traditional MMP14 sensors owing to the m-AuNP-based spherical dual-probe strategy. This sensor has been successfully applied for the detection of exosomal MMP14 from cell culture media and real serum samples. Levels of both MMP14-E and MMP14-A increase in serum from cancer patients, indicating their potential applications as biomarkers in liquid biopsy for disease diagnosis and real-time surveillance.

Graphical abstract: Simultaneous quantification of exosomal MMP14 expression and proteolysis activity on a spherical dual-probe-based fluorescent nanosensor

Supplementary files

Article information

Article type
Paper
Submitted
10 May 2023
Accepted
01 Jun 2023
First published
20 Jun 2023

Analyst, 2023,148, 3483-3490

Simultaneous quantification of exosomal MMP14 expression and proteolysis activity on a spherical dual-probe-based fluorescent nanosensor

S. Yin, A. Chen, X. Fang, P. Zhang and C. Yang, Analyst, 2023, 148, 3483 DOI: 10.1039/D3AN00749A

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