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A Simple Bioconjugate Attachment Protocol for Use in Single Molecule Force Spectroscopy Experiments Based on Mixed Self-Assembled Monolayers |
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| Authors: | Simon J Attwood Anna M C Simpson Rachael Stone SamirW Hamaia Debdulal Roy RichardW Farndale Myriam Ouberai Mark E Welland |
| Affiliation: | 1.Nanoscience Centre, Department of Engineering, Cambridge University, Cambridge, CB3 0FF, UK; E-Mails: (S.J.A.); (M.O.);2.Department of Biochemistry, Cambridge University, Cambridge, CB2 1QW, UK; E-Mails: (A.M.C.S.); (R.S.); (S.W.H.); (R.W.F.);3.National Physical Laboratory, Teddington, TW11 0LW, UK; E-Mail: |
| 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 |