Single Cycle Selection of CD63-targeting Aptamers Using a Microscale Electrophoretic Filtration Device

Aptamers, which show properties of specific binding to their target molecules, are expected to become an alternative to antibodies for use as molecular recognition probes instead of antibodies. However, long and laborious selection procedures have hindered their application. To overcome the drawback...

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Bibliographic Details
Published inBunseki kagaku Vol. 72; no. 3; pp. 111 - 116
Main Authors TOBITA, Arisa, TAKAO, Junku, ENDO, Tatsuro, HISAMOTO, Hideaki, SUEYOSHI, Kenji
Format Journal Article
LanguageEnglish
Japanese
Published Tokyo Japan Science and Technology Agency 01.01.2023
Subjects
Antibodies
Binding
Electrophoresis
Elution
Filtration
Gene sequencing
Hydrogels
Polyacrylamide
Titration calorimetry
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Summary:Aptamers, which show properties of specific binding to their target molecules, are expected to become an alternative to antibodies for use as molecular recognition probes instead of antibodies. However, long and laborious selection procedures have hindered their application. To overcome the drawbacks, rapid and simple selection methods using a microscale electrophoretic filtration device have been developed. In the first study, the aptamers for immunoglobulin E were successfully selected by three cycle selection based on the developed method. To realize more rapid and simple selection, aptamer selection by single cycle procedure was demonstrated. It was confirmed that CD63 as a model target was successfully trapped by the capillary device partially plugged with a polyacrylamide hydrogel. A DNA library containing various sequences were then electrokinetically introduced into the device after trapping CD63, a large volume of DNAs bound with the trapped CD63. After washing and elution schemes, DNAs located in the eluents were enhanced by PCR, and then their sequences were analyzed by next generation sequencer. As a result, one of the obtained aptamer candidates shows similar motifs to the previously reported aptamer. Isothermal titration calorimetry verified the binding ability and specificity of the obtained candidate to CD63. Consequently, it was clarified that the developed method and device is applicable to the rapid and simple selection of aptamers using the single cycle procedure.
ISSN:0525-1931
DOI:10.2116/bunsekikagaku.72.111