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

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 mono...

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Autores principales: Attwood, Simon J., Simpson, Anna M. C., Stone, Rachael, Hamaia, SamirW., Roy, Debdulal, Farndale, RichardW., Ouberai, Myriam, Welland, Mark E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Molecular Diversity Preservation International (MDPI) 2012
Materias:
Article
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497339/
https://www.ncbi.nlm.nih.gov/pubmed/23202965
http://dx.doi.org/10.3390/ijms131013521
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author Attwood, Simon J.
Simpson, Anna M. C.
Stone, Rachael
Hamaia, SamirW.
Roy, Debdulal
Farndale, RichardW.
Ouberai, Myriam
Welland, Mark E.
author_facet Attwood, Simon J.
Simpson, Anna M. C.
Stone, Rachael
Hamaia, SamirW.
Roy, Debdulal
Farndale, RichardW.
Ouberai, Myriam
Welland, Mark E.
author_sort Attwood, Simon J.
collection PubMed
description 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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spelling pubmed-34973392012-11-29 A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers Attwood, Simon J. Simpson, Anna M. C. Stone, Rachael Hamaia, SamirW. Roy, Debdulal Farndale, RichardW. Ouberai, Myriam Welland, Mark E. Int J Mol Sci Article 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. Molecular Diversity Preservation International (MDPI) 2012-10-19 /pmc/articles/PMC3497339/ /pubmed/23202965 http://dx.doi.org/10.3390/ijms131013521 Text en © 2012 by the authors; licensee Molecular Diversity Preservation International, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0 This article is an open-access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0).
spellingShingle Article
Attwood, Simon J.
Simpson, Anna M. C.
Stone, Rachael
Hamaia, SamirW.
Roy, Debdulal
Farndale, RichardW.
Ouberai, Myriam
Welland, Mark E.
A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers
title A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers
title_full A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers
title_fullStr A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers
title_full_unstemmed A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers
title_short A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers
title_sort simple bioconjugate attachment protocol for use in single molecule force spectroscopy experiments based on mixed self-assembled monolayers
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3497339/
https://www.ncbi.nlm.nih.gov/pubmed/23202965
http://dx.doi.org/10.3390/ijms131013521
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