Picomolar affinity antibodies from a fully synthetic naive library selected and evolved by ribosome display.
Nat Biotechnol
; 18(12): 1287-92, 2000 Dec.
Article
em En
| MEDLINE
| ID: mdl-11101809
ABSTRACT
Here we applied ribosome display to in vitro selection and evolution of single-chain antibody fragments (scFvs) from a large synthetic library (Human Combinatorial Antibody Library; HuCAL) against bovine insulin. In three independent ribosome display experiments different clusters of closely related scFvs were selected, all of which bound the antigen with high affinity and specificity. All selected scFvs had affinity-matured up to 40-fold compared to their HuCAL progenitors, by accumulating point mutations during the ribosome display cycles. The dissociation constants of the isolated scFvs were as low as 82 pM, which validates the design of the naïve library and the power of this evolutionary method. We have thus mimicked the process of antibody generation and affinity maturation with a synthetic library in a cell-free system in just a few days, obtaining molecules with higher affinities than most natural antibodies.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Ribossomos
/
RNA Mensageiro
/
Fragmentos de Imunoglobulinas
/
Evolução Molecular Direcionada
/
Biblioteca de Peptídeos
/
Afinidade de Anticorpos
Limite:
Animals
Idioma:
En
Revista:
Nat Biotechnol
Assunto da revista:
BIOTECNOLOGIA
Ano de publicação:
2000
Tipo de documento:
Article
País de afiliação:
Suíça