Cargando…

Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device

This study reports a novel aptamer selection method based on microscale electrophoretic filtration. Aptamers are versatile materials that recognize specific targets and are attractive for their applications in biosensors, diagnosis, and therapy. However, their practical applications remain scarce du...

Descripción completa

Detalles Bibliográficos
Autores principales: Takao, Junku, Nagai, Reina, Endo, Tatsuro, Hisamoto, Hideaki, Sueyoshi, Kenji
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9505737/
https://www.ncbi.nlm.nih.gov/pubmed/36144553
http://dx.doi.org/10.3390/molecules27185818
_version_ 1784796547703111680
author Takao, Junku
Nagai, Reina
Endo, Tatsuro
Hisamoto, Hideaki
Sueyoshi, Kenji
author_facet Takao, Junku
Nagai, Reina
Endo, Tatsuro
Hisamoto, Hideaki
Sueyoshi, Kenji
author_sort Takao, Junku
collection PubMed
description This study reports a novel aptamer selection method based on microscale electrophoretic filtration. Aptamers are versatile materials that recognize specific targets and are attractive for their applications in biosensors, diagnosis, and therapy. However, their practical applications remain scarce due to issues with conventional selection methods, such as complicated operations, low-efficiency separation, and expensive apparatus. To overcome these drawbacks, a selection method based on microscale electrophoretic filtration using a capillary partially filled with hydrogel was developed. The electrophoretic filtration of model target proteins (immunoglobulin E (IgE)) using hydrogel, the electrokinetic injection of DNAs to interact with the trapped proteins, the elimination of DNAs with weak interactions, and the selective acquisition of aptamer candidates with strong interactions were successfully demonstrated, revealing the validity of the proposed concept. Two aptamer candidates for IgE were obtained after three selection cycles, and their affinity for the target was confirmed to be less than 1 nM based on their dissociation constant (K(D)) values. Therefore, the proposed method allows for the selection of aptamers with simple operations, highly effective separation based on electrophoresis and filtration, and a relatively cheap apparatus with disposable devices.
format Online
Article
Text
id pubmed-9505737
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-95057372022-09-24 Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device Takao, Junku Nagai, Reina Endo, Tatsuro Hisamoto, Hideaki Sueyoshi, Kenji Molecules Article This study reports a novel aptamer selection method based on microscale electrophoretic filtration. Aptamers are versatile materials that recognize specific targets and are attractive for their applications in biosensors, diagnosis, and therapy. However, their practical applications remain scarce due to issues with conventional selection methods, such as complicated operations, low-efficiency separation, and expensive apparatus. To overcome these drawbacks, a selection method based on microscale electrophoretic filtration using a capillary partially filled with hydrogel was developed. The electrophoretic filtration of model target proteins (immunoglobulin E (IgE)) using hydrogel, the electrokinetic injection of DNAs to interact with the trapped proteins, the elimination of DNAs with weak interactions, and the selective acquisition of aptamer candidates with strong interactions were successfully demonstrated, revealing the validity of the proposed concept. Two aptamer candidates for IgE were obtained after three selection cycles, and their affinity for the target was confirmed to be less than 1 nM based on their dissociation constant (K(D)) values. Therefore, the proposed method allows for the selection of aptamers with simple operations, highly effective separation based on electrophoresis and filtration, and a relatively cheap apparatus with disposable devices. MDPI 2022-09-08 /pmc/articles/PMC9505737/ /pubmed/36144553 http://dx.doi.org/10.3390/molecules27185818 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Takao, Junku
Nagai, Reina
Endo, Tatsuro
Hisamoto, Hideaki
Sueyoshi, Kenji
Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device
title Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device
title_full Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device
title_fullStr Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device
title_full_unstemmed Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device
title_short Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device
title_sort aptamer selection based on microscale electrophoretic filtration using a hydrogel-plugged capillary device
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9505737/
https://www.ncbi.nlm.nih.gov/pubmed/36144553
http://dx.doi.org/10.3390/molecules27185818
work_keys_str_mv AT takaojunku aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice
AT nagaireina aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice
AT endotatsuro aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice
AT hisamotohideaki aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice
AT sueyoshikenji aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice