Staining‐free gel electrophoresis‐based multiplex enzyme assay using DNA and peptide dual‐functionalized gold nanoparticles

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.

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