Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometry
Research on the effective attachment of aptamers to beads, which is essential for using aptamers, has made relatively little progress. Here, we demonstrate a new method based on flow cytometry to determine the optimal aptamer-to-bead ratio for aptamer immobilization. The fluorescence intensity incre...
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| Main Authors: | , , , , , |
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| Format: | Article |
| Language: | English |
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Wiley
2023-01-01
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| Series: | Scientifica |
| Online Access: | http://dx.doi.org/10.1155/2023/5842652 |
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| _version_ | 1832559689948200960 |
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| author | Sun Young Lee Eun-Ok Kim Daehyuk Jang Soonjae Hwang Ki-Jong Rhee Miyong Yun |
| author_facet | Sun Young Lee Eun-Ok Kim Daehyuk Jang Soonjae Hwang Ki-Jong Rhee Miyong Yun |
| author_sort | Sun Young Lee |
| collection | DOAJ |
| description | Research on the effective attachment of aptamers to beads, which is essential for using aptamers, has made relatively little progress. Here, we demonstrate a new method based on flow cytometry to determine the optimal aptamer-to-bead ratio for aptamer immobilization. The fluorescence intensity increased with a gradual two-fold increase in the aptamer fluorescence concentration, peaked at an aptamer-to-bead ratio of 2.56 × 105, and tended to decrease at higher ratios. A similar pattern was observed in an additional analysis using fluorescence microscopy. However, measurement of the free aptamer concentration after the aptamer-bead conjugation reaction revealed a large aptamer loss compared to the 1.28 × 105 aptamer-bead ratio. In addition, the binding efficiency of the aptamer/bead to the target was highest at the aptamer-to-bead ratio of 1.28 × 105. Taken together, our data suggest that the proposed method is the best and easiest for determining the optimal aptamer-to-bead ratio. |
| format | Article |
| id | doaj-art-4e217d480cdd42339351ffa96dc5b3ce |
| institution | Kabale University |
| issn | 2090-908X |
| language | English |
| publishDate | 2023-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Scientifica |
| spelling | doaj-art-4e217d480cdd42339351ffa96dc5b3ce2025-02-03T01:29:28ZengWileyScientifica2090-908X2023-01-01202310.1155/2023/5842652Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow CytometrySun Young Lee0Eun-Ok Kim1Daehyuk Jang2Soonjae Hwang3Ki-Jong Rhee4Miyong Yun5Lab of Functional AptamerMedical Science Research CenterLab of Functional AptamerDepartment of BiochemistryDepartment of Biomedical Laboratory ScienceLab of Functional AptamerResearch on the effective attachment of aptamers to beads, which is essential for using aptamers, has made relatively little progress. Here, we demonstrate a new method based on flow cytometry to determine the optimal aptamer-to-bead ratio for aptamer immobilization. The fluorescence intensity increased with a gradual two-fold increase in the aptamer fluorescence concentration, peaked at an aptamer-to-bead ratio of 2.56 × 105, and tended to decrease at higher ratios. A similar pattern was observed in an additional analysis using fluorescence microscopy. However, measurement of the free aptamer concentration after the aptamer-bead conjugation reaction revealed a large aptamer loss compared to the 1.28 × 105 aptamer-bead ratio. In addition, the binding efficiency of the aptamer/bead to the target was highest at the aptamer-to-bead ratio of 1.28 × 105. Taken together, our data suggest that the proposed method is the best and easiest for determining the optimal aptamer-to-bead ratio.http://dx.doi.org/10.1155/2023/5842652 |
| spellingShingle | Sun Young Lee Eun-Ok Kim Daehyuk Jang Soonjae Hwang Ki-Jong Rhee Miyong Yun Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometry Scientifica |
| title | Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometry |
| title_full | Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometry |
| title_fullStr | Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometry |
| title_full_unstemmed | Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometry |
| title_short | Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometry |
| title_sort | method to determine the optimal aptamer to bead ratio by using flow cytometry |
| url | http://dx.doi.org/10.1155/2023/5842652 |
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