OligoHydrogelArray (OHA) for Parallelized Solid‐Phase Extraction of Oligonucleotides
Abstract Immobilization of oligonucleotides on solid surfaces is an important step in many experimental workflows in biology, such as gene expression analysis, genotyping, and drug discoveries. Capturing oligonucleotides in a highly efficient miniaturized format is still challenging. In this work, p...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley-VCH
2023-07-01
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| Series: | Advanced Materials Interfaces |
| Subjects: | |
| Online Access: | https://doi.org/10.1002/admi.202300227 |
| _version_ | 1797771372043173888 |
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| author | Michelle J. Iwohn Maximilian Seifermann Patrick Reiser Julius Höpfner Razan El Khaled El Faraj Stefan Heißler Anna A. Popova Pavel A. Levkin |
| author_facet | Michelle J. Iwohn Maximilian Seifermann Patrick Reiser Julius Höpfner Razan El Khaled El Faraj Stefan Heißler Anna A. Popova Pavel A. Levkin |
| author_sort | Michelle J. Iwohn |
| collection | DOAJ |
| description | Abstract Immobilization of oligonucleotides on solid surfaces is an important step in many experimental workflows in biology, such as gene expression analysis, genotyping, and drug discoveries. Capturing oligonucleotides in a highly efficient miniaturized format is still challenging. In this work, preparation of functionalized miniaturized acrylamide hydrogels on a droplet microarray (DMA) chip is reported. By further modification of these hydrogels through the Cu‐catalyzed alkyne‐azide cycloaddition, oligonucleotides are attached covalently, creating the OligoHydrogelArray (OHA). OHA enables the hybridization of oligonucleotides with defined sequences in a parallelized, miniaturized, and high‐throughput manner with an overall loading capacity and extraction efficiency suitable for most biological and chemical applications. This system enables the parallelized solid‐phase extraction of target oligonucleotides out of a mixture for purification and enrichment of DNA/RNA/PNA for application in fields such as transcriptomics. In the current study, are establish the chemical procedure for reproducible preparation of OHA and determine key characteristics, such as height, diameter, and shape of miniaturized gel pads as well as the loading capacity of oligonucleotides within the material. Finally, the ability to extract complementary oligonucleotide strands out of solution through hybridization is demonstrated to set the course for future applications. |
| first_indexed | 2024-03-12T21:36:34Z |
| format | Article |
| id | doaj.art-86fd5797d1954aeda5421c27ceb8fafd |
| institution | Directory Open Access Journal |
| issn | 2196-7350 |
| language | English |
| last_indexed | 2024-03-12T21:36:34Z |
| publishDate | 2023-07-01 |
| publisher | Wiley-VCH |
| record_format | Article |
| series | Advanced Materials Interfaces |
| spelling | doaj.art-86fd5797d1954aeda5421c27ceb8fafd2023-07-27T06:27:26ZengWiley-VCHAdvanced Materials Interfaces2196-73502023-07-011021n/an/a10.1002/admi.202300227OligoHydrogelArray (OHA) for Parallelized Solid‐Phase Extraction of OligonucleotidesMichelle J. Iwohn0Maximilian Seifermann1Patrick Reiser2Julius Höpfner3Razan El Khaled El Faraj4Stefan Heißler5Anna A. Popova6Pavel A. Levkin7Institute of Biological and Chemical Systems‐Functional Molecular Systems Karlsruhe Institute of Technology Hermann‐von‐Helmholtz‐Platz 1 76344 Eggenstein‐Leopoldshafen GermanyInstitute of Biological and Chemical Systems‐Functional Molecular Systems Karlsruhe Institute of Technology Hermann‐von‐Helmholtz‐Platz 1 76344 Eggenstein‐Leopoldshafen GermanyInstitute of Nanotechnology Karlsruhe Institute of Technology Hermann‐von‐Helmholtz‐Platz 1 76344 Eggenstein‐Leopoldshafen GermanyInstitute of Biological and Chemical Systems‐Functional Molecular Systems Karlsruhe Institute of Technology Hermann‐von‐Helmholtz‐Platz 1 76344 Eggenstein‐Leopoldshafen GermanyInstitute of Biological and Chemical Systems‐Functional Molecular Systems Karlsruhe Institute of Technology Hermann‐von‐Helmholtz‐Platz 1 76344 Eggenstein‐Leopoldshafen GermanyInstitute of Functional Interfaces Karlsruhe Institute of Technology Hermann‐von‐Helmholtz‐Platz 1 76344 Eggenstein‐Leopoldshafen GermanyInstitute of Biological and Chemical Systems‐Functional Molecular Systems Karlsruhe Institute of Technology Hermann‐von‐Helmholtz‐Platz 1 76344 Eggenstein‐Leopoldshafen GermanyInstitute of Biological and Chemical Systems‐Functional Molecular Systems Karlsruhe Institute of Technology Hermann‐von‐Helmholtz‐Platz 1 76344 Eggenstein‐Leopoldshafen GermanyAbstract Immobilization of oligonucleotides on solid surfaces is an important step in many experimental workflows in biology, such as gene expression analysis, genotyping, and drug discoveries. Capturing oligonucleotides in a highly efficient miniaturized format is still challenging. In this work, preparation of functionalized miniaturized acrylamide hydrogels on a droplet microarray (DMA) chip is reported. By further modification of these hydrogels through the Cu‐catalyzed alkyne‐azide cycloaddition, oligonucleotides are attached covalently, creating the OligoHydrogelArray (OHA). OHA enables the hybridization of oligonucleotides with defined sequences in a parallelized, miniaturized, and high‐throughput manner with an overall loading capacity and extraction efficiency suitable for most biological and chemical applications. This system enables the parallelized solid‐phase extraction of target oligonucleotides out of a mixture for purification and enrichment of DNA/RNA/PNA for application in fields such as transcriptomics. In the current study, are establish the chemical procedure for reproducible preparation of OHA and determine key characteristics, such as height, diameter, and shape of miniaturized gel pads as well as the loading capacity of oligonucleotides within the material. Finally, the ability to extract complementary oligonucleotide strands out of solution through hybridization is demonstrated to set the course for future applications.https://doi.org/10.1002/admi.202300227droplet microarrayshigh‐throughputminiaturized hydrogelsoligonucleotidesparallelizationsolid‐phase extraction |
| spellingShingle | Michelle J. Iwohn Maximilian Seifermann Patrick Reiser Julius Höpfner Razan El Khaled El Faraj Stefan Heißler Anna A. Popova Pavel A. Levkin OligoHydrogelArray (OHA) for Parallelized Solid‐Phase Extraction of Oligonucleotides Advanced Materials Interfaces droplet microarrays high‐throughput miniaturized hydrogels oligonucleotides parallelization solid‐phase extraction |
| title | OligoHydrogelArray (OHA) for Parallelized Solid‐Phase Extraction of Oligonucleotides |
| title_full | OligoHydrogelArray (OHA) for Parallelized Solid‐Phase Extraction of Oligonucleotides |
| title_fullStr | OligoHydrogelArray (OHA) for Parallelized Solid‐Phase Extraction of Oligonucleotides |
| title_full_unstemmed | OligoHydrogelArray (OHA) for Parallelized Solid‐Phase Extraction of Oligonucleotides |
| title_short | OligoHydrogelArray (OHA) for Parallelized Solid‐Phase Extraction of Oligonucleotides |
| title_sort | oligohydrogelarray oha for parallelized solid phase extraction of oligonucleotides |
| topic | droplet microarrays high‐throughput miniaturized hydrogels oligonucleotides parallelization solid‐phase extraction |
| url | https://doi.org/10.1002/admi.202300227 |
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