Jump to main content
Jump to site search


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

Author affiliations

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

Back to tab navigation

Supplementary files

Publication details

The article was received on 19 Nov 2018, accepted on 17 Jan 2019 and first published on 17 Jan 2019


Article type: Paper
DOI: 10.1039/C8AY02519C
Citation: Anal. Methods, 2019, Advance Article

  •   Request permissions

    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, Advance Article , DOI: 10.1039/C8AY02519C

Search articles by author

Spotlight

Advertisements