Aptabody-aptatope interactions in aptablotting assays

Nanoscale
Simranjeet Singh SekhonYang-Hoon Kim

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.

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Citations

Jan 17, 2020·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Rudi LiuYingfu Li
May 18, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Myeong-Sub SongYang-Hoon Kim
Nov 17, 2020·Frontiers in Chemistry·Yao WangHanyang Yu
Mar 20, 2018·ACS Combinatorial Science·Woo-Ri ShinYang-Hoon Kim
Jul 17, 2018·ACS Combinatorial Science·Se Hee LeeJi-Young Ahn

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