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Nucleic Acids Research, Vol 25, Issue 24 5132-5134, Copyright © 1997 by Oxford University Press

ARTICLES

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

M He and MJ Taussig
Laboratory of Molecular Recognition and Translocus Therapeutics Ltd, The Babraham Institute, Babraham, Cambridge CB2 4AT, UK. mingyue.he@bbsrc.ac.uk

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 (VH/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.
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