Oligonucleotide-Directed Mutagenesis of Antibody Combining Sites
作者:
SharonJacqueline,
Ying Y.Chiou,
SompuramSeshi Reddy,
期刊:
International Reviews of Immunology
(Taylor Available online 1993)
卷期:
Volume 10,
issue 2-3
页码: 113-127
ISSN:0883-0185
年代: 1993
DOI:10.3109/08830189309061689
出版商: Taylor&Francis
关键词: p-azophenylarsonate;transfection;antibody specificity;antibody affinity;idiotype;structure-function
数据来源: Taylor
摘要:
We review here our attempts to achieve a better understanding of the structure—function relationship of antibody combining sites, and to gain insights into the engineering of antibodies with desired specificity and affinity. We have focused on a model system—antibodies to the hapten p-azophenylarsonate (Ars) derived from A/J mice. Oligonucleotide-directed mutagenesis was used to alter the sequence of the variable region genes of such anti-Ars antibodies. Mutant antibodies were generated in hybridoma cells following transfection of the altered genes, and the effects of the primary structure changes on antibody specificity, affinity, and idiotypic expression were assessed. These studies suggest that an antibody combining site with basic specificity for an antigen could be created by introducing a set of a few amino acid residues in the complementarity determining regions, and that the affinity of such a site could be improved one substitution at a time in a sequential manner.
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