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...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2022-09-01
|
| Series: | Molecules |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1420-3049/27/18/5818 |
| _version_ | 1797484373408219136 |
|---|---|
| author | Junku Takao Reina Nagai Tatsuro Endo Hideaki Hisamoto Kenji Sueyoshi |
| author_facet | Junku Takao Reina Nagai Tatsuro Endo Hideaki Hisamoto Kenji Sueyoshi |
| author_sort | Junku Takao |
| collection | DOAJ |
| 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 (<i>K</i><sub>D</sub>) 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. |
| first_indexed | 2024-03-09T23:03:31Z |
| format | Article |
| id | doaj.art-fbf2d39fb20e48ce9153c037be1949fc |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-09T23:03:31Z |
| publishDate | 2022-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-fbf2d39fb20e48ce9153c037be1949fc2023-11-23T17:59:24ZengMDPI AGMolecules1420-30492022-09-012718581810.3390/molecules27185818Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary DeviceJunku Takao0Reina Nagai1Tatsuro Endo2Hideaki Hisamoto3Kenji Sueyoshi4Department of Applied Chemistry, Graduate School of Engineering, Osaka Metropolitan University, Osaka 545-0051, JapanDepartment of Applied Chemistry, Graduate School of Engineering, Osaka Metropolitan University, Osaka 545-0051, JapanDepartment of Applied Chemistry, Graduate School of Engineering, Osaka Metropolitan University, Osaka 545-0051, JapanDepartment of Applied Chemistry, Graduate School of Engineering, Osaka Metropolitan University, Osaka 545-0051, JapanDepartment of Applied Chemistry, Graduate School of Engineering, Osaka Metropolitan University, Osaka 545-0051, JapanThis 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 (<i>K</i><sub>D</sub>) 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.https://www.mdpi.com/1420-3049/27/18/5818aptamer selectionmicroscale electrophoretic filtrationsystematic evolution of ligands by exponential enrichment (SELEX) |
| spellingShingle | Junku Takao Reina Nagai Tatsuro Endo Hideaki Hisamoto Kenji Sueyoshi Aptamer Selection Based on Microscale Electrophoretic Filtration Using a Hydrogel-Plugged Capillary Device Molecules aptamer selection microscale electrophoretic filtration systematic evolution of ligands by exponential enrichment (SELEX) |
| 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 | aptamer selection microscale electrophoretic filtration systematic evolution of ligands by exponential enrichment (SELEX) |
| url | https://www.mdpi.com/1420-3049/27/18/5818 |
| work_keys_str_mv | AT junkutakao aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice AT reinanagai aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice AT tatsuroendo aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice AT hideakihisamoto aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice AT kenjisueyoshi aptamerselectionbasedonmicroscaleelectrophoreticfiltrationusingahydrogelpluggedcapillarydevice |