Rapid One-Step Selection Method for Generating Nucleic Acid Aptamers: Development of a DNA Aptamer against alpha-Bungarotoxin

Background: Nucleic acids based therapeutic approaches have gained significant interest in recent years towards the development of therapeutics against many diseases. Recently, research on aptamers led to the marketing of Macugen (R), an inhibitor of vascular endothelial growth factor (VEGF) for the treatment of age related macular degeneration (AMD). Aptamer technology may prove useful as a therapeutic alternative against an array of human maladies. Considering the increased interest in aptamer technology globally that rival antibody mediated therapeutic approaches, a simplified selection, possibly in one-step, technique is required for developing aptamers in limited time period

Aptamer sequences obtained from the selected clones.

<p>Only the sequences from the random regions are shown.</p

Aptamer characterization.

<p>A. Binding specificity of the aptamer obtained from clone 24 & 51 by surface plasmon resonance (SPR) technique using Biacore 3000. Various concentrations of α-bungarotoxin (1, 10, 20, 30, and 40 µM) were passed through the aptamer which was immobilized on a streptavidin coated sensor chip. The obtained sensorgrams demonstrate the aptamer binding to α-bungarotoxin; B. Predicted structure of the obtained aptamer using the DNA folding platform from mfold web server <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0041702#pone.0041702-Berezovski1" target="_blank">[26]</a>.</p