In　this　article,　we　report　the　first　graphene　oxide　(GO)-based　platform　to　detect　protease　activity　in　a　homogeneous　real-time　format.　In　designing　such　GO-based　biosensing　platform,　we　put　a　protease　substrate　peptide　as　the　linker　between　the　energy　transfer　donor　(QDs)　and　the　energy　transfer　acceptor　(GO)　to　fabricate　the　GO-peptide-QDs　nanoprobes.　In　the　nanoprobes,　the　photoluminescence　(PL)　of　donor　QDs　was　strongly　quenched　due　to　the　presence　of　GO　in　close　proximity.　The　protease　activity　caused　modulation　in　the　efficiency　of　the　energy　transfer　between　the　acceptor　and　donor,　thus　enabling　the　protease　assay.　The　proposed　GO-based　platform　is　easy　to　assemble　and　has　little　background　interference,　yet　still　give　superior　sensitivity　and　rapid　response.　Furthermore,　this　GO-QDs　architecture　can　serve　as　a　universal　platform　by　simply　changing　the　types　of　peptide　sequences　for　the　different　proteases.　In　this　work,　GO-based　platform　has　been　successfully　applied　in　the　sensitive　detection　of　matrix　metalloproteinase　(MMP)　and　thrombin　activity.　Meanwhile,　we　also　utilized　this　platform　to　monitor　the　protease　inhibitor.　The　proposed　GO-based　platform　is　anticipated　to　find　applications　in　the　diagnosis　of　protease-related　diseases　and　screening　of　potential　drugs　with　high　sensitivity　in　a　high-throughput　way.　(C)　2011　Elsevier　B.V.　All　rights　reserved.