title: [Cell-ELA-based determination of binding affinity of DNA aptamer against U87-EGFRvIII cell].
abstract: A15, a DNA aptamer with binding specificity for U87 glioma cells stably overexpressing the epidermal growth factor receptor variant III (U87-EGFRvIII), was generated by cell systematic evolution of ligands by exponential enrichment (cell-SELEX) using a random nucleotide library. Subsequently, we established a cell enzyme-linked assay (cell-ELA) to detect the affinity of A15 compared to an EGFR antibody. We used A15 as a detection probe and cultured U87-EGFRvIII cells as targets. Our data indicate that the equilibrium dissociation constants (K(d)) for A15 were below 100 nmol/L and had similar affinity compared to an EGFR antibody for U87-EGFRvIII. We demonstrated that the cell-ELA was a useful method to determine the equilibrium dissociation constants (K(d)) of aptamers generated by cell-SELEX.
authors.affiliation: Department ofNeurobiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Tan
authors.forename: Huiyu
authors.initials: H
authors.lastname: Liang
authors.forename: Xidong
authors.initials: X
authors.lastname: Wu
authors.forename: Yubo
authors.initials: Y
authors.lastname: Gao
authors.forename: Xingmei
authors.initials: X
authors.lastname: Zhang
mesh_terms: D052157:Aptamers, Nucleotide / Q000378:metabolism*; D001932:Brain Neoplasms / Q000378:metabolism; D045744:Cell Line, Tumor; D066246:ErbB Receptors / Q000235:genetics* / Q000378:metabolism; D005910:Glioma / Q000378:metabolism / Q000473:pathology*; D006801:Humans; D009154:Mutation; D011485:Protein Binding; D052156:SELEX Aptamer Technique / Q000379:methods
chemical_list: D052157:Aptamers, Nucleotide; D066246:ErbB Receptors