An Enzyme-Linked Aptamer Sorbent Assay to Evaluate Aptamer Binding
Nucleic acid aptamers are a class of alternative ligands increasingly growing in importance in the face of contemporary detection challenges. Aptamers offer multiple advantages over traditional ligands like antibodies; however, their ability to specifically bind target molecules must first be confirmed after their generation. Use of a plate-based enzyme-linked aptamer sorbent assay (ELASA) is a generally rapid way to screen and characterize aptamer binding to protein targets. ELASA involves directly plating a protein target onto a nonspecific (polystyrene) surface and assessing binding of functionalized (biotinylated) aptamers to those plated proteins using an enzyme conjugate that recognizes the aptamers. Here, we describe an ELASA that was designed and used to evaluate and compare binding of ssDNA aptamers against the capsids of different strains of human norovirus.
Key wordsAptamer Plate assay Binding evaluation Enzyme-linked aptamer sorbent assay DNA-protein binding Aptamer binding
This work was supported by the Agriculture and Food Research Initiative Competitive Grant no. 2011-68003-30395 from the US Department of Agriculture, National Institute of Food and Agriculture through the NoroCORE project. The authors would like to thank R. Atmar (Baylor College of Medicine, Houston, TX) for providing the VLPs and S. Rupp for his input on the figures.
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