Selection of DNA aptamers against VEGF165 using a protein competitor and the aptamer blotting method.
Two DNA aptamers against a tumor marker protein, human vascular endothelial growth factor (VEGF(165)) were identified. In the screening process, another protein was used as the competitor to isolate those aptamers that have high specificity for the target. In addition, we evaluated the affinities of the enriched library by means of aptamer blotting. The isolated aptamers bound to VEGF(165) with a K(d) value in the range of a few hundred nanomoles, and did not bind to the competitor. This selection method enabled us to efficiently select the specific aptamers against the target protein. These specific aptamers would be useful sensor elements for cancer diagnosis.
Hasegawa
Hijiri
H
Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, Japan.
Sode
Koji
K
Ikebukuro
Kazunori
K
D052157:Aptamers, Nucleotide / Q000302:isolation & purification* / Q000378:metabolism*; D001483:Base Sequence; D001667:Binding, Competitive; D014408:Biomarkers, Tumor / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000378:metabolism; D015723:Gene Library; D006801:Humans; D015336:Molecular Probe Techniques; D009690:Nucleic Acid Conformation; D052156:SELEX Aptamer Technique / Q000379:methods*; D020349:Surface Plasmon Resonance; D042461:Vascular Endothelial Growth Factor A / Q000378:metabolism*
D052157:Aptamers, Nucleotide; D014408:Biomarkers, Tumor; D004800:Enzymes, Immobilized; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A