title: A simple bioconjugate attachment protocol for use in single molecule force spectroscopy experiments based on mixed self-assembled monolayers.
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.
authors.affiliation: Nanoscience Centre, Department of Engineering, Cambridge University, Cambridge, CB3 0FF, UK. mew10@eng.cam.ac.uk.
authors.forename: Simon J
authors.initials: SJ
authors.lastname: Attwood
authors.forename: Anna M C
authors.initials: AM
authors.lastname: Simpson
authors.forename: Rachael
authors.initials: R
authors.lastname: Stone
authors.forename: Samir W
authors.initials: SW
authors.lastname: Hamaia
authors.forename: Debdulal
authors.initials: D
authors.lastname: Roy
authors.forename: Richard W
authors.initials: RW
authors.lastname: Farndale
authors.forename: Myriam
authors.initials: M
authors.lastname: Ouberai
authors.forename: Mark
authors.initials: M
authors.lastname: E Welland
mesh_terms: D001360:Avidin / Q000737:chemistry* / Q000378:metabolism; D001710:Biotin / Q000737:chemistry* / Q000378:metabolism; D006046:Gold / Q000737:chemistry; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D058525:Quartz Crystal Microbalance Techniques; D013499:Surface Properties
chemical_list: D001360:Avidin; D001710:Biotin; D006046:Gold