Guangxi Key Laboratory for Preclinical and Translational Research On Bone and Joint Degenerative Diseases, Affiliated Hospital of Youjiang Medical University for Nationalities, Baise, 533000, Guangxi, China.
West Guangxi Key Laboratory for Prevention and Treatment of High-Incidence Diseases, Youjiang Medical University for Nationalities, Baise, 533000, Guangxi, China.
Mikrochim Acta. 2024 Oct 14;191(11):665. 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)dppz] 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.
基质金属蛋白酶-2（MMP-2）在抗衰老研究中发挥着关键作用。开发具有高特异性、灵敏度和可及性的 MMP-2 高级检测平台至关重要。本研究引入了一种新型的用于 MMP-2 测定的电化学发光（ECL）生物传感器，利用 CRISPR/Cas13a 系统和指数扩增反应（EXPAR）。该生物传感器通过利用 T7 RNA 聚合酶在 MMP-2 相互作用时从 DNA 模板转录 RNA 来工作。这种 RNA 激活 Cas13a，导致信号放大和 ECL 检测。“光开关”分子[Ru（phen）dppz]的掺入通过消除对广泛的电极修饰和清洁的需求，简化了该过程。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。该平台表现出优异的选择性、重现性和稳定性，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中具有广阔的应用前景。
