Earticle

초록

영어

Matrix metalloproteinases (MMPs) have been shown to be abundant in pathological situations such as tumor tissue, osteoarthritis (OA) and rheumatoid arthritis (RA). The extent of MMPs detected in biological samples provides important information for diagnosis, prognosis, and therapy monitoring of diseases relating with MMP. A number of attempts have been made to detect activities of MMPs in biological samples and to evaluate the effects of MMP inhibitors. Previously, we have demonstrated that a highly efficient MMP fluorogenic probe, which is a pair consisting of a fluorophore (Cy5.5) and a quencher (BHQ-3), can detect MMP-13 in OA-induced rat model with a low background and significant signal recovery, enabling a highly sensitive visual detection in vitro and in vivo. In this study, we developed a new high-throughput MMP detection kit based on a 96-well microplate by immobilizing a rationale designed chitosan-conjugated fluorogenic probe and investigated whether this new MMP detection kit can monitor MMP activity. Glycol chitosan (GC), natural polymer, was used as a linker to immobilize and amplify MMP fluorogenic probe on a 96-well microplate for a highly sensitive detection. When MMP enzymes were simply added and incubated in a 96-well microplate immobilized with GCconjugated MMP probe, the fluorescence of each well was recovered and the fluorescence intensity showed distinct difference within minutes through Kodak Image Station. Furthermore, recovery of fluorescent signals in the MMP detection kit was proportional to concentrations of immobilized GCconjugated MMP probe and, importantly, treated concentrations of pure MMP enzymes. The detection kit is most specific for MMPs, compared with other enzymes including caspase-3 and 20s proteasome. Additionally, the detection kit was used to detect MMP activity in biological samples such as synovial fluids from OA patients and showed the fluorescence intensity to correlate with the stage of OA development. The detection kit may be applicable in detecting MMP activity in other various biological samples and evaluate the effects of MMP inhibitors in a rapid and easy fashion.

키워드

저자

• Ju Hee RYU [ Biomedical Research Center, KIST, Seoul, 130-650. ]
• Aeju LEE [ Biomedical Research Center, KIST, Seoul, 130-650. ]
• Seulki LEE [ Biomedical Research Center, KIST, Seoul, 130-650. ]
• Ick Chan KWON [ Biomedical Research Center, KIST, Seoul, 130-650. ]
• In-Chan YOUN [ Biomedical Research Center, KIST, Seoul, 130-650. ]
• Kuiwon CHOI [ Biomedical Research Center, KIST, Seoul, 130-650. ]
• Kwangmeyung KIM [ Biomedical Research Center, KIST, Seoul, 130-650. ]

참고문헌

발행기관

간행물

표지보기 관심저널 등록

• 간행물명

  한국생물공학회 학술대회

• 간기

  반년간

• 수록기간

  1985~2013

• 십진분류

  KDC 476 DDC 576