Immobilization-Free Determination of Dissociation Constants Independent of Ligand Size Using MicroScale Thermophoresis.
Methods Mol Biol
; 2570: 129-140, 2023.
Article
em En
| MEDLINE
| ID: mdl-36156779
ABSTRACT
The quantitative characterization of aptamer-ligand interactions is an important step in the aptamer development process. However, certain pitfalls impede KD determination, especially when working with small molecule ligands. These include altered binding behavior caused by ligand immobilization. Further, the compulsory requirement for major differences in size between the bound and unbound state makes small molecule ligands ineligible for separation-based methods. MicroScale Thermophoresis circumvents such limitations as binding is accurately quantified with both binding partners free in solution and independent of ligand size. In this chapter, we present a protocol for the characterization of a DNA aptamer binding to its small molecule ligand daunorubicin.
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Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Aptâmeros de Nucleotídeos
Idioma:
En
Revista:
Methods Mol Biol
Assunto da revista:
BIOLOGIA MOLECULAR
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
Alemanha