On-chip assay of matrix metalloproteinase-3 activity using fluorescence-conjugated gelatin arrays

Deok Hoon Kong, Se Hui Jung, Seung Taek Lee, Kwon Soo Ha

Research output: Contribution to journalArticlepeer-review

6 Citations (Scopus)

Abstract

We developed a new high-throughput approach to measure matrix metalloproteinase (MMP)-3 activity using Cy3-conjugated gelatin arrays. We fabricated Cy3-conjugated gelatin arrays by immobilizing Cy3-conjugated gelatin onto well-type amine arrays. The ratio of Cy3-NHS to gelatin was optimized to improve the efficiency of the arrays. MMP-3 activity was determined by the decrease of fluorescence intensity due to MMP-3-mediated degradation of Cy3-conjugated gelatin. The gelatinolytic activity of MMP-3 increased in the presence of Brij-35, a non-ionic detergent, in a dose-dependent manner. Cy3-conjugated gelatin arrays were successfully applied for the analyses of MMP-3 activity, which showed a dose-dependent degradation of the Cy3-conjugated substrate. This chip-based activity assay using a Cy3-conjugated gelatin array is simple, rapid, cost-effective, and has a potential for activity-based proteomic researches and screening of MMP-3 inhibitors.

Original languageEnglish
Pages (from-to)210-216
Number of pages7
JournalBiochip Journal
Volume4
Issue number3
DOIs
Publication statusPublished - 2010

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Biomedical Engineering
  • Electrical and Electronic Engineering

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