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Antibody recognition imaging by force microscopy

Nature Biotechnology volume 17, pages 901–905 (1999)Cite this article

Abstract

We have developed a method that combines dynamic force microscopy with the simultaneous molecular recognition of an antigen by an antibody, during imaging. A magnetically oscillated atomic force microscopy tip carrying a tethered antibody was scanned over a surface to which lysozyme was bound. By oscillating the probe at an amplitude of only a few nanometers, the antibody was kept in close proximity to the surface, allowing fast and efficient antigen recognition and gentle interaction between tip and sample. Antigenic sites were evident from reduction of the oscillation amplitude, as a result of antibody–antigen recognition during the lateral scan. Lysozyme molecules bound to the surface were recognized by the antibody on the scanning tip with a few nanometers lateral resolution. In principle, any ligand can be tethered to the tip; thus, this technique could potentially be used for nanometer-scale epitope mapping of biomolecules and localizing receptor sites during biological processes.

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Figure 1: (A) Multilayer topography image. Image size was 500 nm.
Figure 2: (A) AFM tip–lysozyme interaction during topography imaging.
Figure 3: (A) Time course of antibody state.

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Acknowledgements

We are grateful to Drs. Hermann J. Gruber and Werner Baumgartner for enlightening discussions. This work was supported by the Austrian Science Foundation projects P12801/2-MED (A.R., D.B., H.S., and P.H.), the Austrian Ministry of Science project GZ200.026/2-Pr/4/98 (A.R., D.B., H.S., and P.H.), the EC-BIOTECHNOLOGY program project ERBBIO4CT960592 (A.R., D.B., H.S., and P.H.), the National Institutes of Health (S.J.S.-G.), and the National Science Foundation project BIR 9513233 (S.M.L.).

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Authors and Affiliations

  1. Institute for Biophysics, University of Linz, Linz, A-4040, Austria

    Anneliese Raab, Dirk Badt, Hansgeorg Schindler & Peter Hinterdorfer

  2. Molecular Imaging Corporation, Phoenix, 85044, 9830A 51st St., Suite A124, AZ

    Wenhai Han

  3. Division of Basic Sciences Frederick Cancer Research and Development Center, Basic Research Laboratory, NCI, NIH, Frederick , 21702-1201, MD

    Sandra J. Smith-Gill

  4. Department of Physics and Astronomy, Arizona State University, Tempe, 85287-1504, AZ

    Stuart M. Lindsay

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  1. Anneliese Raab
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  2. Wenhai Han
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  4. Sandra J. Smith-Gill
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  6. Hansgeorg Schindler
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  7. Peter Hinterdorfer
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Correspondence to Peter Hinterdorfer.

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Raab, A., Han, W., Badt, D. et al. Antibody recognition imaging by force microscopy. Nat Biotechnol 17, 901–905 (1999). https://doi.org/10.1038/12898

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