Cell-ELA-based determination of binding affinity of DNA aptamer against U87-EGFRvIII cell.
- Author:
Yan TAN
1
;
Huiyu LIANG
;
Xidong WU
;
Yubo GAO
;
Xingmei ZHANG
Author Information
1. Department ofNeurobiology, School of Basic Medical Sciences, Southern Medical University, Guangzhou 510515, Guangdong, China.
- Publication Type:Journal Article
- MeSH:
Aptamers, Nucleotide;
metabolism;
Brain Neoplasms;
metabolism;
Cell Line, Tumor;
Glioma;
metabolism;
pathology;
Humans;
Mutation;
Protein Binding;
Receptor, Epidermal Growth Factor;
genetics;
metabolism;
SELEX Aptamer Technique;
methods
- From:
Chinese Journal of Biotechnology
2013;29(5):664-671
- CountryChina
- Language:Chinese
-
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.
