Cell-ELA-based determination of binding affinity of DNA aptamer against U87-EGFRvIII cell / 生物工程学报
Chinese Journal of Biotechnology
; (12): 664-671, 2013.
Article
en Zh
| WPRIM
| ID: wpr-233211
Biblioteca responsable:
WPRO
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.
Texto completo:
1
Índice:
WPRIM
Asunto principal:
Patología
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Unión Proteica
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Neoplasias Encefálicas
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Línea Celular Tumoral
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Aptámeros de Nucleótidos
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Técnica SELEX de Producción de Aptámeros
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Receptores ErbB
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Genética
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Glioma
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Metabolismo
Límite:
Humans
Idioma:
Zh
Revista:
Chinese Journal of Biotechnology
Año:
2013
Tipo del documento:
Article