Use of a novel mutagenesis strategy, optimized residue substitution, to decrease the off-rate of an anti-gp120 antibody.
Mol Immunol
; 32(14-15): 1065-72, 1995 Oct.
Article
en En
| MEDLINE
| ID: mdl-8544856
ABSTRACT
We have developed a novel strategy to decrease the antibodyantigen off-rate which we call optimized residue substitution. This strategy employs alanine substitution to first identify residues non-optimal for binding, as evidenced by a decrease in off-rate upon alanine replacement. These positions are then individually randomized to all amino acids, and the best replacement for each position determined. Finally, a construct which combines all optimized substitutions is generated and evaluated. We applied this strategy to the heavy chain CDR3 of P5Q, a scFv antibody which recognizes an epitope on the V3 loop of HIV gp120. We identified two amino acid substitutions that together decrease the off-rate by nearly ten-fold. The contributions by the two substitutions were near additive, indicative of independent affects on binding. We suggest that this strategy can be generalized to strengthen proteinligand and proteinprotein interactions in other systems.
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Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
Anticuerpos Anti-VIH
/
Proteína gp120 de Envoltorio del VIH
/
Mutagénesis Insercional
/
Afinidad de Anticuerpos
Tipo de estudio:
Clinical_trials
/
Prognostic_studies
Idioma:
En
Revista:
Mol Immunol
Año:
1995
Tipo del documento:
Article
País de afiliación:
Estados Unidos