Method to Determine the Optimal Aptamer-to-Bead Ratio by Using Flow Cytometry.

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 × 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

SUBMITTER: Lee SY 

PROVIDER: S-EPMC10353897 | biostudies-literature | 2023

REPOSITORIES: biostudies-literature