Antibody-ribosome-mRNA (ARM) complexes as efficient selection particles for in vitro display and evolution of antibody combining sites

被引：197

作者：

He, MY ^{[1
]}

Taussig, MJ ^{[1
]}

机构：

[1] Therapeut Ltd, Babraham Inst, Lab Mol Recognit & Translocus, Cambridge CB2 4AT, England

来源：

NUCLEIC ACIDS RESEARCH | 1997年 / 25卷 / 24期

关键词：

D O I：

10.1093/nar/25.24.5132

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a rapid, eukaryotic, in vitro method for selection and evolution of antibody combining sites using antibody-ribosome-mRNA (ARM) complexes as selection particles, ARMs carrying single-chain (V-H/K) binding fragments specific for progesterone were selected using antigen-coupled magnetic beads; selection simultaneously captured the genetic information as mRNA, making it possible to generate and amplify cDNA by single-step RT-PCR on the ribosome-bound mRNA for further manipulation, Using mutant libraries, antigen-binding ARMs were enriched by a factor of 10(4)-10(5)-fold in a single cycle, with further enrichment in repeated cycles, While demonstrated here for antibodies, the method has the potential to be applied equally for selection of receptors or peptides from libraries.

引用

页码：5132 / 5134

页数：3

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