| Info. | Vol.7 - No.2 (2013.06.20) |
|---|---|
| Title | Development of microbiochip for detection of metalloproteinase 7 using fluorescence resonance energy transfer |
| Authors | Seung Yong Lee1, Byoungwook Han2, Chansoo Park1, Je-Sik Jeong1, Jeong Jin Ahn1, Seung-Mo Ha2, Seung Yong Hwang1 & Yoomin Ahn2 |
| Institutions | 1Department of Bio-Nanotechnology, Graduate School,
Hanyang University, Ansan, Gyeonggi-do, Korea 2Department of Mechanical Engr. Hanyang University, Ansan, Gyeonggi-do, Korea |
| Abstract | A protease is any enzyme that catalyzes the hydrolysis of proteins into smaller peptide fragments and amino acids, a process known as proteolysis. They are involved in a multitude of normal biological processes as well as in diseases, including cancer, stroke and infections. Here we present a microfluidicbased assay system to detect proteolytic activity using fluorescence resonance energy transfer (FRET) by quantum dot (QD)-peptide conjugates immobilized on microbeads. As an energy donor, QD was immobilized on the microbead surface by the avidin-biotin interaction. As an energy acceptor, the fluorophorelabeled peptide was then associated with QD, thus quenching the photoluminescence (PL) of the QD. The functionalized microbeads were introduced into the microbiochip and captured by a micropillar in the reaction chamber. In the presence of matrix metalloprotease- 7 (MMP-7) as a model protease, the PL of QD quenched by fluorophore was recovered due to the proteolytic activity of MMP-7 in the fabricated icrobiochip. Moreover, the FRET efficiency induced by MMP-7 was linearly dependent on the logarithmic concentration of MMP-7. This technology is not limited to sensing MMP-7, but could be used to monitor other protease activities (Schematic diagram). |
| Keyword | Microbiochip, Matrix Metalloproteinase 7, Fluorescence Resonance Energy Transfer (FRET), Quantum Dot (QD), Bead-based assay, Enzyme assay |
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