title: Staining-free gel electrophoresis-based multiplex enzyme assay using DNA and peptide dual-functionalized gold nanoparticles.
abstract: We report a simple staining-free gel electrophoresis method to simultaneously probe protease and nuclease. Utilizing gold nanoparticles (Au-NPs) dual-functionalized with DNA and peptide, the presence and concentration of nuclease and protease are determined concurrently from the relative position and intensity of the bands in the staining-free gel electrophoresis. The use of Au-NPs eliminates the need for staining processes and enables naked eye detection, while a mononucleotide-mediated approach facilitates the synthesis of DNA/peptide conjugated Au-NPs and simplifies the operation procedures. Multiplex detection and quantification of DNase I and trypsin are successfully demonstrated.
authors.affiliation: Department of Chemical and Biomolecular Engineering, The Hong Kong University of Science and Technology, Kowloon, Hong Kong SAR, China.
authors.forename: Wenting
authors.initials: W
authors.lastname: Zhao
authors.forename: Chunlei
authors.initials: C
authors.lastname: Yao
authors.forename: Xiaoteng
authors.initials: X
authors.lastname: Luo
authors.forename: Li
authors.initials: L
authors.lastname: Lin
authors.forename: I-Ming
authors.initials: IM
authors.lastname: Hsing
mesh_terms: D004247:DNA / Q000737:chemistry* / Q000378:metabolism; D003850:Deoxyribonuclease I / Q000032:analysis / Q000378:metabolism; D004586:Electrophoresis / Q000379:methods*; D057075:Enzyme Assays / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D055767:Immobilized Proteins / Q000737:chemistry / Q000378:metabolism; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism; D014357:Trypsin / Q000032:analysis / Q000378:metabolism
chemical_list: D055767:Immobilized Proteins; D010455:Peptides; D006046:Gold; D004247:DNA; D003850:Deoxyribonuclease I; D014357:Trypsin