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Molecules 2016, 21(4), 421; doi:10.3390/molecules21040421

Methods for Improving Aptamer Binding Affinity

Department of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan
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Academic Editors: Alain O. A. Miller and Jean Jacques Vanden Eynde
Received: 21 December 2015 / Revised: 6 March 2016 / Accepted: 22 March 2016 / Published: 28 March 2016
(This article belongs to the Special Issue Aptamers: Past, Present, and Future)
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Abstract

Aptamers are single stranded oligonucleotides that bind a wide range of biological targets. Although aptamers can be isolated from pools of random sequence oligonucleotides using affinity-based selection, aptamers with high affinities are not always obtained. Therefore, further refinement of aptamers is required to achieve desired binding affinities. The optimization of primary sequences and stabilization of aptamer conformations are the main approaches to refining the binding properties of aptamers. In particular, sequence optimization using combined in silico sequence recombinations and in vitro functional evaluations is effective for the improvement of binding affinities, however, the binding affinities of aptamers are limited by the low hydrophobicity of nucleic acids. Accordingly, introduction of hydrophobic moieties into aptamers expands the diversity of interactions between aptamers and targets. Moreover, construction of multivalent aptamers by connecting aptamers that recognize distinct epitopes is an attractive approach to substantial increases in binding affinity. In addition, binding affinities can be tuned by optimizing the scaffolds of multivalent constructs. In this review, we summarize the various techniques for improving the binding affinities of aptamers. View Full-Text
Keywords: aptamer; binding affinity; aptamer sequence; unnatural nucleotide; multivalent binding aptamer; binding affinity; aptamer sequence; unnatural nucleotide; multivalent binding
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Hasegawa, H.; Savory, N.; Abe, K.; Ikebukuro, K. Methods for Improving Aptamer Binding Affinity. Molecules 2016, 21, 421.

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