Dunaway Adam B, Sullivan Richard S, Siegel Katherine J, Milam Valeria T
School of Materials Science and Engineering, Georgia Institute of Technology, 771 Ferst Dr. NW, Atlanta, Georgia 30332.
School of Materials Science and Engineering, Georgia Institute of Technology, 771 Ferst Dr. NW, Atlanta, Georgia 30332; Wallace C. Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 313 Ferst Dr., Atlanta, Georgia 30332; and Petit Institute of Bioengineering and Bioscience, Georgia Institute of Technology, 315 Ferst Dr., Atlanta, Georgia 30332.
Biointerphases. 2015 Mar 18;10(1):019015. doi: 10.1116/1.4915107.
In the current study, the authors quantify the binding activity of particle-immobilized DNA aptamers to their nucleotide and non-nucleotide targets. For the purposes of this work, DNA and vascular endothelial growth factor (VEGF) binding analysis was carried out for VEGF-binding aptamers and compared to that of an ampicillin-binding aptamer as well as a non-aptamer DNA probe. Binding analysis followed incubation of one target type, coincubation of both DNA and VEGF targets, and serial incubations of each target type. Moreover, recovery of aptamer binding activity following displacement of the DNA target from aptamer:DNA duplexes was also explored. Flow cytometry served as the quantitative tool to directly monitor binding events of both the DNA target and protein target to the various aptamer and non-aptamer functionalized particles. The current work demonstrates how processing steps such as annealing and binding history of particle-immobilized aptamers can affect subsequent binding activity. To this end, the authors demonstrate the ability to fully recover DNA target binding activity capabilities and to partially recover protein target binding activity.
