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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Bibliographic Details
Published in:Scientifica (Cairo) 2023, Vol.2023, p.5842652-8
Main Authors: Lee, Sun Young, Kim, Eun-Ok, Jang, Daehyuk, Hwang, Soonjae, Rhee, Ki-Jong, Yun, Miyong
Format: Article
Language:English
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Summary: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.
ISSN:2090-908X
2090-908X
DOI:10.1155/2023/5842652