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

Mikrochim Acta. 2024 Oct 14;191(11):665. doi: 10.1007/s00604-024-06707-4.

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.

Keywords: CRISPR/Cas13a; Electrochemiluminescence; MMP-2; T7 RNA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biosensing Techniques* / methods
  • CRISPR-Cas Systems*
  • Electrochemical Techniques* / methods
  • Humans
  • Limit of Detection*
  • Luminescent Measurements* / methods
  • Matrix Metalloproteinase 2* / genetics
  • Matrix Metalloproteinase 2* / metabolism
  • Nucleic Acid Amplification Techniques / methods
  • Reproducibility of Results

Substances

  • Matrix Metalloproteinase 2
  • MMP2 protein, human