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Issue 22, 2017
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Aptabody–aptatope interactions in aptablotting assays

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Abstract

We demonstrate an aptablotting assay method that involves direct and indirect aptabody recognition. Nanoscale single-stranded DNA aptamers against GST and DIG-tags are utilized as aptabodies (GST-2 and DIG-1, respectively), and the GST-2 aptabody binding site, or aptatope, as predicted by a MOE-docking simulation of the protein–aptamer complex, shows the interaction of the GST-2 aptabody at the catalytically active region. The aptabody–aptatope interaction was evaluated by an in vitro enzyme inhibitory analysis. The binding capacity of the GST-2 aptabody was assessed by dot-blot, EMSA and SDS-PAGE/electroblot analyses, and the results showed that the aptabodies interact with both the native mono-/dimeric form and the denatured GST form on a membrane. The use of aptabodies can overcome the obstacles of current immunoblot assays, and these molecules are easily assessable via ELISA systems. Moreover, the hybridization of aptabodies and antibodies (hybrid-aptablotting) may have considerable impacts on the design of bioassay platforms.

Graphical abstract: Aptabody–aptatope interactions in aptablotting assays

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Publication details

The article was received on 15 Mar 2017, accepted on 27 Apr 2017 and first published on 03 May 2017


Article type: Paper
DOI: 10.1039/C7NR01827D
Citation: Nanoscale, 2017,9, 7464-7475
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    Aptabody–aptatope interactions in aptablotting assays

    S. S. Sekhon, H. Um, W. Shin, S. Lee, J. Min, J. Ahn and Y. Kim, Nanoscale, 2017, 9, 7464
    DOI: 10.1039/C7NR01827D

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