Issue 8, 2019

Force spectroscopic detection of peptide cleavage by thrombin exploiting biotin–streptavidin interactions in a bio-sensing context

Abstract

Atomic-force-microscopy-based single-molecule force spectroscopy (AFM-SMFS) has become an important technique as the basis of novel, label-free biosensing strategies. In this work, we explored the possibility of using AFM-SMFS to detect the specific peptide cleavage activity by thrombin proteases. To achieve this aim, an oligopeptide with a sequence that is recognized and cleaved by thrombin was spanned between an AFM tip and a solid surface via a PEG-based linker system including the stable biotin–streptavidin receptor/ligand complex. We found that force spectral signatures associated with the biotin–streptavidin unbinding in the absence of thrombin can be clearly distinguished from the signatures of spectra collected after thrombin-induced peptide cleavage. However, this is possible only in a well-defined window of peak force and tip–sample separation distance values, in which specific streptavidin–biotin interactions do not overlap with non-specific interactions among all the other system components. The results were employed to engineer a sensing architecture capable of detecting the presence of thrombin at concentrations higher than about 0.2 μM.

Graphical abstract: Force spectroscopic detection of peptide cleavage by thrombin exploiting biotin–streptavidin interactions in a bio-sensing context

Supplementary files

Article information

Article type
Paper
Submitted
19 Nov 2018
Accepted
17 Jan 2019
First published
17 Jan 2019

Anal. Methods, 2019,11, 1102-1110

Force spectroscopic detection of peptide cleavage by thrombin exploiting biotin–streptavidin interactions in a bio-sensing context

J. Li, Q. Li, S. Potthoff, G. Wei and C. C. Lucio, Anal. Methods, 2019, 11, 1102 DOI: 10.1039/C8AY02519C

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