Coordination-Based Site-Specific Labeling Strategy for Electrogenerated Chemiluminescence Biosensing of Matrix Metalloproteinase 2

Abstract

Matrix metalloproteinase 2 (MMP-2) is an important biomarker for some diseases. Herein, one first-case coordination-based site-specific labeling strategy is proposed for electrogenerated chemiluminescence (ECL) biosensing of MMP-2 by employing an iridium(III) solvent complex as a signal reagent and a histidine (His)-containing peptide as a molecular recognition substrate. One ECL probe was prepared via coordination labeling of the His-containing peptide with one iridium(III) solvent complex ([(3-(2-pyridyl)benzoic acid)₂Ir(DMSO)Cl], Ir1-DMSO). High ECL efficiency and good cleavage ability by MMP-2 were obtained for the ECL probe. By combining the high sensitivity of the ECL method, the good specificity of the peptide, and the simpleness of the magnetic bead-based assay, one “cleavage-magnetic enrichment type” ECL biosensing method was developed to detect MMP-2. MMP-2 can be sensitively detected in the linear range of 1.0–10 ng/mL with a limit of quantification of 1.0 ng/mL and a limit of detection of 0.3 ng/mL. Moreover, the ECL biosensing method was successfully applied for the determination of MMP-2 in serum samples with recoveries from 98.0% ± 8.0% to 108.0% ± 6.0%. Further, high affinity (K_d = 0.11 nM) was obtained for the Ir1-DMSO-labeled His-containing peptide and MMP-2. This work may pave the way for the labeling of His-containing biomolecules with an iridium(III) solvent complex and provides a promising method in point-of-care testing of MMP-2.