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International journal of molecular sciences
Oct 19, 2012;13 (10): 13521-41.

A simple bioconjugate attachment protocol for use in single molecule force spectroscopy experiments based on mixed self-assembled monolayers.

Simon J Attwood, Anna M C Simpson, Rachael Stone, Samir W Hamaia, Debdulal Roy, Richard W Farndale, Myriam Ouberai, Mark E Welland
Author Information
  1. Simon J Attwood: Nanoscience Centre, Department of Engineering, Cambridge University, Cambridge, CB3 0FF, UK. mew10@eng.cam.ac.uk.
PMID: 23202965 DOI: 10.3390/ijms131013521

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.

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Grants

  1. G0500707/Medical Research Council

MeSH Term

Avidin
Biotin
Gold
Microscopy, Atomic Force
Protein Binding
Quartz Crystal Microbalance Techniques
Surface Properties

Chemicals

Avidin
Biotin
Gold
Journal Article Research Support, Non-U.S. Gov't
Article has an altmetric score of 3

Word Cloud

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