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Journal of Biomolecular Screening
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1087057106293841v1
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Article

Obligate Multivalent Recognition of Cell Surface Tomoregulin following Selection from a Multivalent Phage Antibody Library

Tara Heitner1, Noboru Satozawa1, Kirk Mclean1, David Vogel1, Ronald R. Cobb1, Bing Liu1, Mithra Mahmoudi1, Silke Finster2, Brent Larsen1, Ying Zhu1, Hongxing Zhou3, Beate Müller-Tiemann2, Felipe Monteclaro1, Xiao-Yan Zhao1, David R. Light1*

1 Berlex Biosciences, Richmond, California
2 Schering AG, Berlin, Germany
3 Amgen Inc., Seattle, Washington

* To whom correspondence should be addressed. E-mail: david_light{at}berlex.com.


  Abstract

A therapeutic antibody candidate (AT-19) isolated using multivalent phage display binds native tomoregulin (TR) as a multimer not as a monomer. This report raises the importance of screening and selecting phage antibodies on native antigen and reemphasizes the possibility that potentially valuable antibodies are discarded when a monomeric phage display system is used for screening. A detailed live cell panning selection and screening method to isolate multivalently active antibodies is described. AT-19 is a fully human antibody recognizing the cell surface protein TR, a proposed prostate cancer target for therapeutic antibody internalization. AT-19 was isolated from a multivalent single-chain variable fragment (scFv) antibody library rescued with hyperphage. The required multivalency for isolation of AT-19 is supported by fluorescence activated cell sorting data demonstrating binding of the multivalent AT-19 phage particles at high phage concentrations and failure of monovalent particles to bind. Pure monomeric scFv AT-19 does not bind native receptor on cells, whereas dimeric scFv or immunoglobulin G binds with nanomolar affinity. The isolation of AT-19 antibody with obligate bivalent binding activity to native TR is attributed to the use of a multivalent display of scFv on phage and the method for selecting and screening by alternate use of 2 recombinant cell lines.

Key Words: antibody phage display, single-chain Fv, scFv, hyperphage, cell panning

First published on November 7, 2006, doi:10.1177/1087057106293841

Journal of Biomolecular Screening 2006;11:985.

A more recent version of this article appeared on December 1, 2006

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D. W. Schneider, T. Heitner, B. Alicke, D. R. Light, K. McLean, N. Satozawa, G. Parry, J. Yoo, J. S. Lewis, and R. Parry
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