GOLD SELEX: a novel SELEX approach for the development of high-affinity aptamers against small molecules without residua
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ORIGINAL PAPER
GOLD SELEX: a novel SELEX approach for the development of high-affinity aptamers against small molecules without residual activity Bandhan Chatterjee 1 & Neeti Kalyani 1 & Anjali Anand 1 & Eshan Khan 2 & Soonjyoti Das 1 & Vipul Bansal 3 & Amit Kumar 2 & Tarun Kumar Sharma 1 Received: 22 April 2020 / Accepted: 29 September 2020 # Springer-Verlag GmbH Austria, part of Springer Nature 2020
Abstract GOLD SELEX, a novel SELEX approach has been developed that obviates the need for target immobilization for aptamer development. The approach purely relies on the affinity of the aptamers towards its target, to get detached from the gold nanoparticle (GNP) surface (weak attraction) after binding with its target. Thus, only the completely detached aptamers are selected for the next round of SELEX. This, in-process, also addresses the issue of residual binding and thus improves the sensitivity of the developed aptamers. As a proof of concept for establishing the utility of the approach for small molecules, we have developed aptamers against dichlorvos (DV), a pesticide in just 8 rounds. Using these aptamer candidates, we have developed an aptamer-NanoZyme (GNP having peroxidase mimic activity) based colorimetric assay. The developed aptamer displayed high affinity (Kd in sub micromolar range) and selectivity for DV. The developed assay could detect as low as 15 μM DV. The best-performing aptamer was also able to work in real samples like river water and commercial apple juice. The GOLD SELEX approach developed in this study, we believe, can act as a template for future SELEX strategy development and can replace the conventional SELEX strategy. Keywords Aptamer . GOLD SELEX . Small-molecule SELEX . Residual binding . NanoZyme
Introduction Nucleic acid aptamers are chemical siblings of antibodies [1]. They are typically generated with a process of in-vitro evolution Neeti Kalyani and Anjali Anand contributed equally to this work. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s00604-020-04577-0) contains supplementary material, which is available to authorized users. * Tarun Kumar Sharma [email protected] 1
Aptamer Technology and Diagnostics Laboratory, Multidisciplinary Clinical and Translational Research Group, Translational Health Science and Technology Institute (THSTI), Faridabad, Haryana 121001, India
2
Discipline of Biosciences and Biomedical Engineering, Indian Institute of Technology Indore, Simrol, Indore 453552, India
3
Ian Potter NanoBioSensing Facility, NanoBiotechnology Research Lab (NBRL), School of Science, RMIT University, GPO Box 2476, Melbourne, Victoria 3001, Australia
known as Systematic Evolution of Ligands by EXponential enrichment (SELEX) [2, 3]. In recent years, this process has been successfully employed to generate a variety of aptamers for various analytes ranging from small molecules to proteins to the whole cells [4–6]. Several SELEX-derived small-molecule binding aptamers evinced target-induced structural switching capabilities
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