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Title: Free-standing three-dimensional graphene scaffolds for protease functional assay
Authors: Ng, Zhi Kai
Yan, Evelias
Goyal, Garima
Gudlur, Sushanth
Kanagavel, Deepankumar
Yıldız, Ümit Hakan
Tsang, Siu Hon
Alagappan, Palaniappan
Teo, Edwin Hang Tong
Keywords: Free standing graphene scaffold
Functional assay
Graphene foam
Publisher: Elsevier
Abstract: Three-dimensional graphene scaffolds (3d-GS) of high porosity possessing good fluorescence quenching properties are potential candidates for the development of optical biosensors. Herein, we demonstrate the feasibility of utilising intact and free-standing 3d-GS for sensitive detection of proteases, a class of disease diagnosis biomarkers of significant interest. Recombinant OmpT was employed as a model protease for validating the proposed methodology. A short (15-residue) peptide sequence encoding a specific recognition site for OmpT was end-labelled with a fluorescent dye (5-FAM) whose fluorescence is quenched when the peptide is anchored to 3d-GS. However, in the presence of OmpT, the peptide is cleaved and released from 3d-GS, resulting in a significant recovery in fluorescence. The functional assay described herein involves a single step fabrication process of anchoring the peptide to 3d-GS. The integrity of the 3d-GS is hypothesised to overcome the concern of dynamic re-quenching associated with the typical homogeneous assays based on graphene, yielding a limit of detection (LOD) of ∼ 140 nM, which is over an order higher than homogeneous assays performed using the same composition of graphene in powdered form. To the best of our knowledge, this is the first report on utilising free-standing 3d-GS for facile assaying of proteases. © 2023 Elsevier B.V.
ISSN: 0925-4005
Appears in Collections:Chemistry / Kimya
Scopus İndeksli Yayınlar Koleksiyonu / Scopus Indexed Publications Collection
WoS İndeksli Yayınlar Koleksiyonu / WoS Indexed Publications Collection

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