Selection and characterization of DNA aptamers against VEGF165 with aptamer blotting method and its application.
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.
Ikebukuro
Kazunori
K
Department of Biotechnology & Life Science, Tokyo University of Agriculture & Technology, 2-24-16 Nakacho, Koganei, Tokyo, 184-8588, Japan. ikebu@cc.tuat.ac.jp
Hasegawa
Hijiri
H
Sode
Koji
K
D052157:Aptamers, Nucleotide / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D001483:Base Sequence; D001667:Binding, Competitive; D014408:Biomarkers, Tumor / Q000378:metabolism*; D006801:Humans; D015336:Molecular Probe Techniques; D008969:Molecular Sequence Data; D009690:Nucleic Acid Conformation; D052156:SELEX Aptamer Technique; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D052157:Aptamers, Nucleotide; D014408:Biomarkers, Tumor; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A