rapid one-step selection method for generating nucleic acid aptamers development of a dna aptamer against α-bungarotoxin快速一步选择方法生成核酸寡核苷酸适配子与α-bungarotoxin dna适体的发展.pdfVIP

Rapid One-Step Selection Method for Generating Nucleic Acid Aptamers: Development of a DNA Aptamer against a-Bungarotoxin 1,2 1 1 3 1 Lasse H. Lauridsen , Hadi A. Shamaileh , Stacey L. Edwards , Elena Taran , Rakesh N. Veedu * 1 School of Chemistry and Molecular Biosciences, The University of Queensland, Brisbane, Queensland, Australia, 2 The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, Hørsholm, Denmark, 3 Australian National Fabrication Facility, Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, Brisbane, Queensland, Australia Abstract 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 MacugenH, 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. Principal Findings: Herein, we present a simple one-step selection of DNA aptamers against a-bungarotoxin. A toxin immobilized glass coverslip was subjected to nucleic acid pool binding and extensive washing followed by PCR enrichment of the selected aptamers. One round of selection successfully identified a DNA aptamer sequence with a binding affinity of