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Nucleic Acids Research 2005 33(4):e35; doi:10.1093/nar/gni033
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Published online 24 February 2005

© The Author 2005. Published by Oxford University Press. All rights reserved
The online version of this article has been published under an open access model. Users are entitled to use, reproduce, disseminate, or display the open access version of this article for non-commercial purposes provided that: the original authorship is properly and fully attributed; the Journal and Oxford University Press are attributed as the original place of publication with the correct citation details given; if an article is subsequently reproduced or disseminated not in its entirety but only in part or as a derivative work this must be clearly indicated. For commercial re-use, please contact journals.permissions{at}oupjournals.org

Methods Online

Selection of functional human antibodies from retroviral display libraries

Johannes H. Urban, Richard M. Schneider, Marta Compte1, Carsten Finger, Klaus Cichutek, Luis Álvarez-Vallina1 and Christian J. Buchholz*

Medizinische Biotechnologie, Paul-Ehrlich-Institut 63225 Langen, Germany 1 Department of Immunology, Hospital Universitario Clinica Puerta de Hierro Madrid, Spain

*To whom correspondence should be addressed. Tel: +49 6103 774011; Fax: +49 6103 771255; Email: bucch{at}pei.de

Received December 13, 2004. Revised February 3, 2005. Accepted February 3, 2005.

Antibody library technology represents a powerful tool for the discovery and design of antibodies with high affinity and specificity for their targets. To extend the technique to the expression and selection of antibody libraries in an eukaryotic environment, we provide here a proof of concept that retroviruses can be engineered for the display and selection of variable single-chain fragment (scFv) libraries. A retroviral library displaying the repertoire obtained after a single round of selection of a human synthetic scFv phage display library on laminin was generated. For selection, antigen-bound virus was efficiently recovered by an overlay with cells permissive for infection. This approach allowed more than 103-fold enrichment of antigen binders in a single selection cycle. After three selection cycles, several scFvs were recovered showing similar laminin-binding activities but improved expression levels in mammalian cells as compared with a laminin-specific scFv selected by the conventional phage display approach. Thus, translational problems that occur when phage-selected antibodies have to be transferred onto mammalian expression systems to exert their therapeutic potential can be avoided by the use of retroviral display libraries.

The authors wish it to be known that, in their opinion, the first two authors should be regarded as joint First Authors


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