Screening and Improvement of an Anti-VEGF DNA Aptamer
Open Access
- 6 January 2010
- Vol. 15 (1), 215-225
- https://doi.org/10.3390/molecules15010215
Abstract
To obtain an aptamer with a high affinity for vascular endothelial growth factor (VEGF), we focused on the receptor-binding domain (RBD) of VEGF as a target epitope. Three rounds of screening gave Vap7, which bound to the VEGF isoforms VEGF121 and VEGF165 with KD values of 1.0 nM and 20 nM, respectively. Moreover, Vap7 showed specificity within the VEGF family. Secondary structure predictions and circular dicrhoism suggested that Vap7 folds into a G-quadruplex structure. We obtained a mutant aptamer that contains only this region of the aptamer sequence. This truncated mutant (V7t1) bound to both VEGF121 and VEGF165 with KD values of 1.1 nM and 1.4 nM, respectively. Its sequence was 5'-TGTGGGGGTGGACGGGCCGGGTAGA-3', and it appeared to form a G-quadruplex structure. We also produced an aptamer heterodimer consisting of our previously derived aptamer (del5-1), which binds to the heparin-binding domain of VEGF, linked to V7t1. The resulting heterodimer bound strongly to VEGF165 with a KD value of 4.7 × 102 pM.Keywords
This publication has 21 references indexed in Scilit:
- On the Signaling of Electrochemical Aptamer‐Based Sensors: Collision‐ and Folding‐Based MechanismsElectroanalysis, 2009
- Electrochemical Sensing of Aptamer‐Protein Interactions Using a Magnetic Particle Assay and Single‐Use Sensor TechnologyElectroanalysis, 2009
- Biophysical characterization of DNA aptamer interactions with vascular endothelial growth factorPeptide Science, 2008
- Immobilized DNA Switches as Electronic Sensors for Picomolar Detection of Plasma ProteinsJournal of the American Chemical Society, 2008
- Isolation of DNA Aptamers for CDP-Ribitol Synthase, and Characterization of Their Inhibitory and Structural PropertiesChemBioChem, 2007
- Characterization and application of aptamers for Taq DNA polymerase selected using an evolution-mimicking algorithmBiotechnology Letters, 2006
- Direct In Vitro Selection of a 2′-O-Methyl Aptamer to VEGFCell Chemical Biology, 2005
- 2′-Fluoropyrimidine RNA-based Aptamers to the 165-Amino Acid Form of Vascular Endothelial Growth Factor (VEGF165)Published by Elsevier BV ,1998
- In vitro selection of RNA molecules that bind specific ligandsNature, 1990
- Systematic Evolution of Ligands by Exponential Enrichment: RNA Ligands to Bacteriophage T4 DNA PolymeraseScience, 1990