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Int. J. Mol. Sci. 2012, 13(10), 13521-13541; doi:10.3390/ijms131013521
Article

A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers

1
, 2
, 2
, 2
, 3
, 2
, 1
 and 1,*
1 Nanoscience Centre, Department of Engineering, Cambridge University, Cambridge, CB3 0FF, UK 2 Department of Biochemistry, Cambridge University, Cambridge, CB2 1QW, UK 3 National Physical Laboratory, Teddington, TW11 0LW, UK
* Author to whom correspondence should be addressed.
Received: 22 August 2012 / Revised: 9 September 2012 / Accepted: 29 September 2012 / Published: 19 October 2012
(This article belongs to the Special Issue Advances in Single Molecule Spectroscopy)
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Abstract

Single molecule force spectroscopy is a technique that can be used to probe the interaction force between individual biomolecular species. We focus our attention on the tip and sample coupling chemistry, which is crucial to these experiments. We utilised a novel approach of mixed self-assembled monolayers of alkanethiols in conjunction with a heterobifunctional crosslinker. The effectiveness of the protocol is demonstrated by probing the biotin-avidin interaction. We measured unbinding forces comparable to previously reported values measured at similar loading rates. Specificity tests also demonstrated a significant decrease in recognition after blocking with free avidin.
Keywords: atomic force microscopy; single molecule force spectroscopy; biotin-avidin interaction; mixed self-assembled monolayers atomic force microscopy; single molecule force spectroscopy; biotin-avidin interaction; mixed self-assembled monolayers
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Attwood, S.J.; Simpson, A.M.C.; Stone, R.; Hamaia, S.W.; Roy, D.; Farndale, R.W.; Ouberai, M.; E.Welland, M. A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers. Int. J. Mol. Sci. 2012, 13, 13521-13541.

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