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Updated: Jul 10, 2026

A Flow Cytometry-Based Cell Surface Protein Binding Assay for Assessing Selectivity and Specificity of an Anticancer Aptamer
Published on: September 13, 2022
Selection and characterization of DNA aptamers against VEGF165 with aptamer blotting method and its application
Kazunori Ikebukuro1, Hijiri Hasegawa, Koji Sode
1Department of Biotechnology & Life Science, Tokyo University of Agriculture & Technology, 2-24-16 Nakacho, Koganei, Tokyo, 184-8588, Japan. ikebu@cc.tuat.ac.jp
Abstract:
We have isolated two DNA aptamers against a tumor marker protein, human vascular endothelial growth factor (VEGF165). To select aptamers that have high specificity for VEGF165, we used another protein as a competitor in the SELEX process. In addition, we evaluated the screened library with aptamer blotting. Isolated aptamers bound to VEGF165 with Kd in a few hundred-nanomolar ranges, and did not bind to a competitor. This selection method allowed us to select the specific aptamers against a target protein efficiently. Furthermore, isolated aptamers did not bind to VEGF121. These specific DNA aptamers against VEGF165 would be a useful sensing element for cancer diagnosis.

