Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometryopen access
- Authors
- Lee, Sun Young; Kim, Eun-Ok; Jang, Daehyuk; Hwang, Soonjae; Rhee, Ki-Jong; Yun, Miyong
- Issue Date
- Jul-2023
- Publisher
- HINDAWI LTD
- Citation
- SCIENTIFICA, v.2023
- Journal Title
- SCIENTIFICA
- Volume
- 2023
- URI
- https://scholarworks.bwise.kr/gachon/handle/2020.sw.gachon/88770
- DOI
- 10.1155/2023/5842652
- ISSN
- 2090-908X
- Abstract
- 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 x 10(5), 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 x 10(5) 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 x 10(5). Taken together, our data suggest that the proposed method is the best and easiest for determining the optimal aptamer-to-bead ratio.
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