Quantitative analysis of the number of antigens immobilized on a glass surface by AFM

Ultramicroscopy
Hyonchol KimAtsushi Ikai

Abstract

To develop force measurements using an atomic force microscope (AFM) in a quantitative manner, it is necessary to estimate the number density of target molecules on a sample surface, and for this, the sensitivity of detection should be known. In this study, the AFM was used as a mechanical detector and an antigen and its antibody were used as a model to evaluate the sensitivity of detection. Antigens were immobilized on a glass surface and number density was estimated by monitoring optical absorbance due to product formation by the reaction of crosslinkers. The concentration of antigen was controlled by mixing control peptides. A microbead was used as a probe and antibodies were immobilized on the bead. AFM force measurements were then made for a range of number densities in the order of 10-10(6) antigen molecules per square micrometer of surface and were compared to evaluate the sensitivity of detection. Our result establishes the reliability of estimating a number of molecules like receptors on the cell surface, and indicates that the AFM is useful as a mechanical detector with high sensitivity.

References

May 1, 1996·Biophysical Journal·U DammerH J Güntherodt
Oct 3, 1999·Neuroscience Letters·S TakigamiM Ichikawa
Sep 14, 2000·Proceedings of the National Academy of Sciences of the United States of America·C GergelyJ Hemmerlé
May 2, 2002·Proceedings of the National Academy of Sciences of the United States of America·Mike McElfreshRobert E Rudd
Jul 12, 2002·The EMBO Journal·Daniel J MüllerGina E Sosinsky

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Citations

Mar 15, 2008·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·Atsushi Ikai
Oct 10, 2007·Colloids and Surfaces. B, Biointerfaces·Hyonchol KimAtsushi Ikai

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