Qianli Tang

An advanced electrochemiluminescence (ECL) biosensor was developed that integrates T7 RNA polymerase amplification, ladder-branch hybridization chain reaction (HCR), and the precise targeting capabilities of CRISPR/Cas13a technology. The novelty of this research lies in the unique combination of these three cutting-edge technologies, which has not been previously utilized together in biosensing platforms, enabling highly sensitive and specific detection of biomolecules with exceptional precision. This innovative biosensor addresses the critical need for sensitive and specific detection of matrix metalloproteinase-2 (MMP-2), a key biomarker in cancer diagnostics. Through meticulous optimization of amplification and reaction conditions, the biosensor demonstrated remarkable sensitivity and specificity, achieving a detection limit as low as 6.34 aM, surpassing existing methodologies. The biosensor also exhibited excellent stability and reproducibility across multiple scans and maintained consistent functionality over an extended period, highlighting its reliability for practical applications. The effectiveness of the biosensor was validated using real samples, demonstrating its capability to accurately quantify MMP-2 in complex biological matrices with high recovery and minimal interference. The integration of isothermal amplification and CRISPR/Cas13a within the ECL biosensor platform represents a significant advancement in molecular diagnostics, offering a powerful tool for real-time monitoring of MMP-2. This combination of technologies sets the platform apart from traditional methods, marking a significant step forward in biosensor innovation. This biosensor holds substantial promise for revolutionizing cancer diagnostics and facilitating personalized treatment strategies, ultimately aiming to improve patient outcomes in cancer care. Future research may explore further enhancements and applications of this biosensor in various clinical settings. Graphical 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