Electrochemiluminescence biosensor for MMP-2 determination using CRISPR/Cas13a and EXPAR amplification: a novel approach for anti-aging research

• Published in: Mikrochimica acta (1966) Vol. 191; no. 11; p. 665
• Main Authors: Tang, Qiang, Wang, Jie, Zhang, Jiayi, Zeng, Hongyu, Su, Zhixue, Zhu, Xiying, Wei, Jihua, Gong, Yuanxun, Tang, Qianli, Zhang, Kai, Liao, Xianjiu
• Format: Journal Article
• Language: English
• Published: Vienna Springer Vienna 01.11.2024; Springer Nature B.V
• Subjects: Aging (metallurgy); Analytical Chemistry; Biological properties; Biosensing Techniques - methods; Biosensors; Characterization and Evaluation of Materials; Chemistry; Chemistry and Materials Science; CRISPR-Cas Systems; Electrochemical Techniques - methods; Electrochemiluminescence; Humans; Limit of Detection; Luminescent Measurements - methods; Matrix Metalloproteinase 2 - genetics; Matrix Metalloproteinase 2 - metabolism; Matrix metalloproteinases; Microengineering; Nanochemistry; Nanotechnology; Nucleic Acid Amplification Techniques - methods; Reproducibility of Results; Ribonucleic acid; RNA; Electrochemiluminescence; MMP-2; T7 RNA; CRISPR/Cas13a
• ISSN: 0026-3672; 1436-5073; 1436-5073
• DOI: 10.1007/s00604-024-06707-4

Matrix metalloproteinase-2 (MMP-2) plays a pivotal role in anti-aging research. Developing advanced detection platforms for MMP-2 with high specificity, sensitivity, and accessibility is crucial. This study introduces a novel electrochemiluminescence (ECL) biosensor for MMP-2 determination, leveraging the CRISPR/Cas13a system and Exponential Amplification Reaction (EXPAR). The biosensor operates by utilizing the T7 RNA polymerase to transcribe RNA from a DNA template upon MMP-2 interaction. This RNA activates Cas13a, leading to signal amplification and ECL detection. The incorporation of the "photoswitch" molecule [Ru(phen) 2 dppz] 2+ streamlines the process by eliminating the need for extensive electrode modification and cleaning. Under optimized conditions, the biosensor achieved an impressive detection limit of 12.8 aM for MMP-2. The platform demonstrated excellent selectivity, reproducibility, and stability, making it highly suitable for detecting MMP-2 in complex biological samples. This innovative approach shows great potential for applications in molecular diagnostics and anti-aging research. Graphical Abstract