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

Berlex Biosciences, Richmond, California

Noboru Satozawa

Berlex Biosciences, Richmond, California

Kirk Mclean

Berlex Biosciences, Richmond, California

David Vogel

Berlex Biosciences, Richmond, California

Ronald R. Cobb

Berlex Biosciences, Richmond, California

Bing Liu

Berlex Biosciences, Richmond, California

Mithra Mahmoudi

Berlex Biosciences, Richmond, California

Silke Finster

Schering AG, Berlin, Germany

Brent Larsen

Berlex Biosciences, Richmond, California

Ying Zhu

Berlex Biosciences, Richmond, California

Hongxing Zhou

Amgen Inc., Seattle, Washington

Beate Müller-Tiemann

Schering AG, Berlin, Germany

Felipe Monteclaro

Berlex Biosciences, Richmond, California

Xiao-Yan Zhao

Berlex Biosciences, Richmond, California

David R. Light

Berlex Biosciences, Richmond, California

A therapeutic antibody candidate (AT-19) isolated using multivalent phage display binds native tomoregulin (TR) as a mul-timer 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

This version was published on December 1, 2006

Journal of Biomolecular Screening, Vol. 11, No. 8, 985-995 (2006)
DOI: 10.1177/1087057106293841


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