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Label-free detection of the aptamer binding on protein patterns using Kelvin probe force microscopy (KPFM)

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Abstract

Anti-lysozyme aptamers are found to preferentially bind to the edge of a tightly packed lysozyme pattern. Such edge-binding is due to the better accessibility and flexibility of the edge lysozyme molecules. Kelvin probe force microscopy (KPFM) was used to study the aptamer–lysozyme binding. Our results show that KPFM is capable of detecting the aptamer–protein binding down to the 30 nm scale. The surface potential of the aptamer–lysozyme complex is approximately 12 mV lower than that of the lysozyme. The surface potential images of the aptamer-bound lysozyme patterns have the characteristic shoulder steps around the pattern edge, which is much wider than that of a clean lysozyme pattern. These results demonstrate the potentials of KPFM as a label-free method for the detection of protein–DNA interactions.

Aptamers preferentially bind on the edge of a protein pattern as revealed by Kelvin force microscopy.

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Acknowledgment

This research is supported by the University of Kentucky faculty start-up grant. We thank Dr. Mehdi M. Yazdanpanah and Dr. Robert W. Cohn from ElectroOptics Research Institute and Nanotechnology Center, University of Louisville for providing us the Ag2Ga needle tip for high-resolution KPFM characterizations.

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Authors and Affiliations

  1. Department of Chemistry, University of Kentucky, Rose Street, Lexington, KY, 40506, USA

    Pei Gao & Yuguang Cai

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  1. Pei Gao
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  2. Yuguang Cai
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Correspondence to Yuguang Cai.

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Gao, P., Cai, Y. Label-free detection of the aptamer binding on protein patterns using Kelvin probe force microscopy (KPFM). Anal Bioanal Chem 394, 207–214 (2009). https://doi.org/10.1007/s00216-008-2577-8

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  • DOI: https://doi.org/10.1007/s00216-008-2577-8

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