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Membrane-based actuation for high-speed single molecule force spectroscopy studies using AFM

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Abstract

Atomic force microscopy (AFM)-based dynamic force spectroscopy of single molecular interactions involves characterizing unbinding/unfolding force distributions over a range of pulling speeds. Owing to their size and stiffness, AFM cantilevers are adversely affected by hydrodynamic forces, especially at pulling speeds >10 μm/s, when the viscous drag becomes comparable to the unbinding/unfolding forces. To circumvent these adverse effects, we have fabricated polymer-based membranes capable of actuating commercial AFM cantilevers at speeds ≥100 μm/s with minimal viscous drag effects. We have used FLUENT®, a computational fluid dynamics (CFD) software, to simulate high-speed pulling and fast actuation of AFM cantilevers and membranes in different experimental configurations. The simulation results support the experimental findings on a variety of commercial AFM cantilevers and predict significant reduction in drag forces when membrane actuators are used. Unbinding force experiments involving human antibodies using these membranes demonstrate that it is possible to achieve bond loading rates ≥106 pN/s, an order of magnitude greater than that reported with commercial AFM cantilevers and systems.

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Acknowledgments

We thank Dr. Fang Kong and Jiangguo Lin for help with data collection. We also thank Dr. Peter Kottke for helpful discussions involving FLUENT®. This work was supported by the National Institutes of Health (AI060799).

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Author notes

  1. Krishna Sarangapani

    Present address: Department of Physiology & Biophysics, University of Washington, Seattle, WA, USA

Authors and Affiliations

  1. Woodruff School of Mechanical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332, USA

    Krishna Sarangapani, Hamdi Torun, Ofer Finkler & Levent Degertekin

  2. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, Atlanta, GA, 30332, USA

    Cheng Zhu

  3. 801, Ferst Drive, Atlanta, GA, 30332, USA

    Levent Degertekin

Authors
  1. Krishna Sarangapani
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  2. Hamdi Torun
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  3. Ofer Finkler
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  4. Cheng Zhu
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  5. Levent Degertekin
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Corresponding author

Correspondence to Levent Degertekin.

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Sarangapani, K., Torun, H., Finkler, O. et al. Membrane-based actuation for high-speed single molecule force spectroscopy studies using AFM. Eur Biophys J 39, 1219–1227 (2010). https://doi.org/10.1007/s00249-009-0575-1

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  • DOI: https://doi.org/10.1007/s00249-009-0575-1

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