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Detection of Protease Activity by Fluorescent Peptide Zymography
Published on: January 20, 2019
Coordination-Based Site-Specific Labeling Strategy for Electrogenerated Chemiluminescence Biosensing of Matrix
Manping Qian1, Hong Huang1, Danyang Zhang1
1Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province, School of Chemistry and Chemical Engineering, Shaanxi Normal University, Xi'an, Shaanxi 710062, P. R. China.
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 (Kd = 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.

