Abstract
Peptide-based electrochemical biosensors have proven to be powerful sensing strategies for the detection of proteases and offer a highly versatile system, as simple tuning of the immobilised substrate peptide leads to a new sensing platform. The most common immobilisation strategy of peptides onto an electrode surface is via a self-assembled monolayer (SAM) through a thiol group that can be readily chemically incorporated in the target peptide. However, the successful application of these platforms in complex biological samples can be frustrated by the lack of stability of the peptide-based SAM, that can lead to false positives and limited shelf-life. Herein, we investigated the stability of a peptide-based electrochemical platform endowed with a single or a tribranched thiol group for attaching onto the electrode surface. Side-by-side comparison demonstrated that the tripod anchor generated a highly robust peptide-based electrochemical biosensor that showed improved stability when compared to the monoanchor analogue, simplifying data analysis and interpretation, while showing efficient protease detection and similar sensing capabilities.
Highlights
AN Self-assembled monolayers (SAMs) have been used for the fabrication of a vast range of M devices encompassing organic electronics [1], molecular motors [2], sensors and actuators [3]
D In our recent work, we reported an electrochemical peptide-based biosensor for trypsin which E consisted of a short peptide sequence, labelled with methylene blue (MB) as a redox reporter T and immobilised onto a gold electrode surface using cysteine as the thiol-containing anchor P [30]
We report on AC the sensing abilities of the new electrochemical biosensor, its stability under a range of conditions, while comparing its analytical performance to that of the benchmark monodentate probe as a control
Summary
AN Self-assembled monolayers (SAMs) have been used for the fabrication of a vast range of M devices encompassing organic electronics [1], molecular motors [2], sensors and actuators [3]. Among the different types of SAMs, thiols on gold, which typically T comprise a sulfhydryl anchoring group attached to a spacer followed by a terminal cargo such P as a specific strand of DNA, an aptamer or peptide, have received much attention. In part, this E is due to their ease of preparation, driven by the readily formed bond between the sulfhydryl C group and the gold surface [6, 7]. Trithiol based-anchors have proven to be suitable for robust bioconjugation onto gold surfaces and have been employed for attaching DNA strands onto
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