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Selection of DNA Aptamers Against Botulinum Neurotoxin E for Development of Fluorescent Aptasensor
Author(s) -
Ren Shuo,
Shin HyeSoo,
Gedi Vinayakumar,
Dua Pooja,
Lee DongKi,
Kim Soyoun
Publication year - 2017
Publication title -
bulletin of the korean chemical society
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.237
H-Index - 59
ISSN - 1229-5949
DOI - 10.1002/bkcs.11085
Subject(s) - aptamer , systematic evolution of ligands by exponential enrichment , clostridium botulinum , chemistry , nucleic acid , dna , combinatorial chemistry , toxoid , computational biology , microbiology and biotechnology , biochemistry , nanotechnology , toxin , biology , rna , virology , gene , materials science , vaccination , tetanus
Botulinum neurotoxins ( BoNTs ) produced by Clostridium botulinum are the deadliest toxins known to mankind with a potential of possible bioterrorism agents. Due to the high‐potential harm of these toxins, a sensitive and real‐time monitoring capable detection systems are required for the early stage detection. In this study, a novel sol–gel‐based SELEX was used to isolate single‐stranded nucleic acids ( ssDNA ) aptamers against purified BoNT type E toxoid ( BoNTE ) using a chemically synthesized random N40 ssDNA library. The bound aptamer sequences after five rounds were analyzed using next‐generation sequencing and the resulted high‐frequency aptamers were characterized using biolayer interferometry. Among the tested, aptamer BT5 .6 showed low nanomolar binding affinity ( K D , 53.3 nM ) toward BoNTE . As a proof of concept, the aptamer was utilized in a graphene oxide‐based detection platform. In summary, we screened and identified a ssDNA aptamer with high affinity toward BoNTE , which, we believe can be used for development of sensitive detection systems.