Ryu, Ju Hee Lee, Aeju Lee, Seulki Ahn, Cheol-Hee Park, Jong Woong Leary, James F. Park, Sangjin Kim, Kwangmeyung Kwon, Ick Chan Youn, In-Chan Choi, Kuiwon 2024-01-20T19:02:01Z 2024-01-20T19:02:01Z 2021-09-02 2010-07 1043-1802 https://pubs.kist.re.kr/handle/201004/131289 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 in 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 or 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. English AMER CHEMICAL SOC RHEUMATOID-ARTHRITIS DNA MICROARRAYS OSTEOARTHRITIS TISSUE DEGRADATION PROGRESSION INHIBITOR CLEAVAGE PROTEASE FLUID "One-Step" Detection of Matrix Metalloproteinase Activity Using a Fluorogenic Peptide Probe-Immobilized Diagnostic Kit Article 10.1021/bc100008b 1 BIOCONJUGATE CHEMISTRY, v.21, no.7, pp.1378 - 1384 BIOCONJUGATE CHEMISTRY 21 7 1378 1384 scie scopus 000280028100030 2-s2.0-77954883421 Biochemical Research Methods Biochemistry & Molecular Biology Chemistry, Multidisciplinary Chemistry, Organic Biochemistry & Molecular Biology Chemistry Article RHEUMATOID-ARTHRITIS DNA MICROARRAYS OSTEOARTHRITIS TISSUE DEGRADATION PROGRESSION INHIBITOR CLEAVAGE PROTEASE FLUID