Affinity maturation by phage display.
Methods Mol Biol
; 525: 309-22, xv, 2009.
Article
en En
| MEDLINE
| ID: mdl-19252854
ABSTRACT
Antibodies are indispensable tools for research, diagnostics, and therapy. However, sometimes antibodies with the most favourable specificity profile lack sufficient affinity for a desired application. Here, we describe a method to increase the affinity of recombinant scFv antibody fragments based on random mutagenesis and phage display under stringent conditions. Random mutations are inserted by performing several rounds of error-prone PCR. After construction of a mutated antibody gene library, affinity selection is performed by panning with washing conditions optimized for off-rate-dependent selection. Alternatively, panning in solution with competition can be used to enrich binders with improved binding properties.
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Biblioteca de Péptidos
/
Biología Molecular
/
Afinidad de Anticuerpos
Límite:
Animals
Idioma:
En
Revista:
Methods Mol Biol
Asunto de la revista:
BIOLOGIA MOLECULAR
Año:
2009
Tipo del documento:
Article
País de afiliación:
Alemania