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

IF 5.3 2区化学 Q1 CHEMISTRY, ANALYTICAL

Microchimica Acta Pub Date : 2024-10-14 DOI:10.1007/s00604-024-06707-4

Qiang Tang, Jie Wang, Jiayi Zhang, Hongyu Zeng, Zhixue Su, Xiying Zhu, Jihua Wei, Yuanxun Gong, Qianli Tang, Kai Zhang, Xianjiu Liao

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Abstract

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)₂dppz]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

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利用 CRISPR/Cas13a 和 EXPAR 扩增技术测定 MMP-2 的电化学发光生物传感器：抗衰老研究的新方法

基质金属蛋白酶-2（MMP-2）在抗衰老研究中起着举足轻重的作用。开发特异性强、灵敏度高、易于使用的先进 MMP-2 检测平台至关重要。本研究利用 CRISPR/Cas13a 系统和指数扩增反应（EXPAR），介绍了一种用于测定 MMP-2 的新型电化学发光（ECL）生物传感器。该生物传感器利用 T7 RNA 聚合酶在 MMP-2 相互作用时从 DNA 模板转录 RNA。这种 RNA 会激活 Cas13a，导致信号放大和 ECL 检测。加入 "光开关 "分子[Ru(phen)2dppz]2+后，无需对电极进行大量修饰和清洁，从而简化了整个过程。在优化条件下，该生物传感器对 MMP-2 的检测限达到了令人印象深刻的 12.8 aM。该平台具有出色的选择性、再现性和稳定性，非常适合检测复杂生物样本中的 MMP-2。这种创新方法在分子诊断和抗衰老研究方面具有巨大的应用潜力。

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来源期刊

Microchimica Acta 化学-分析化学

CiteScore

9.80

自引率

5.30%

发文量

410

审稿时长

2.7 months

期刊介绍： As a peer-reviewed journal for analytical sciences and technologies on the micro- and nanoscale, Microchimica Acta has established itself as a premier forum for truly novel approaches in chemical and biochemical analysis. Coverage includes methods and devices that provide expedient solutions to the most contemporary demands in this area. Examples are point-of-care technologies, wearable (bio)sensors, in-vivo-monitoring, micro/nanomotors and materials based on synthetic biology as well as biomedical imaging and targeting.