"One-step" detection of matrix metalloproteinase activity using a fluorogenic peptide probe-immobilized diagnostic kit

Ju Hee Ryu, Aeju Lee, Seulki Lee, Cheol H. Ahn, Jong Woong Park, James F. Leary, Sangjin Park, Kwang Meyung Kim, Ick Chan Kwon, In Chan Youn, Kuiwon Choi

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Abstract

Matrix metalloproteinases (MMPs) have been shown to be abundant in pathological conditions such as cancer, osteoarthritis (OA), and rheumatoid arthritis (RA). The extent of MMPs detected in biological samples provides important clinical information for diagnosis, prognosis, and therapeutic monitoring of various diseases relating with MMPs. Herein, we developed a new high-throughput MMP diagnostic kit (MMP-D-KIT) based on a 96-well plate by immobilizing MMP-13 specific fluorogenic peptide probes (MMP peptide probe), which is a pair consisting of a near-infrared (NIR) fluorophore (Cy5.5) and a quencher (BHQ-3), onto the biocompatible glycol chitosan (GC) polymer anchored 96-well plate. When MMP enzymes were simply added and incubated in a MMP-D-KIT, the fluorescence of each well was recovered and the fluorescence intensity showed distinct difference within minutes through NIR fluorescence imaging system. The fluorescence was recovered not only by MMP-13 activity, but also by other MMPs activity. Furthermore, recovery of NIR fluorescent signals in MMP-D-KIT was proportional to concentrations of immobilized MMP peptide probe-GC conjugates and, importantly, MMP concentration. The MMP-D-KIT is most specific for target MMP, compared with other enzymes including caspase-3 and 20s proteasome. Additionally, the MMP-D-KIT was used to detect MMP activity in biological samples such as synovial fluid from 12 OA patients (grades 1-4 based on the Kellgren-Lawrence grading scale). It was found that the fluorescence intensity measured using MMP-D-KIT decidedly correlates with the progression of OA. The MMP-D-KIT could be applicable in detecting MMP activities in various biological samples and evaluating the effects of MMP inhibitors in a rapid and easy fashion.

Original languageEnglish
Pages (from-to)1378-1384
Number of pages7
JournalBioconjugate Chemistry
Volume21
Issue number7
DOIs
Publication statusPublished - 2010 Jul 21

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ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Organic Chemistry
  • Biomedical Engineering
  • Pharmaceutical Science
  • Pharmacology

Cite this

Ryu, J. H., Lee, A., Lee, S., Ahn, C. H., Park, J. W., Leary, J. F., Park, S., Kim, K. M., Kwon, I. C., Youn, I. C., & Choi, K. (2010). "One-step" detection of matrix metalloproteinase activity using a fluorogenic peptide probe-immobilized diagnostic kit. Bioconjugate Chemistry, 21(7), 1378-1384. https://doi.org/10.1021/bc100008b