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Obligate multivalent recognition of cell surface tomoregulin following selection from a multivalent phage antibody library.
- Source :
-
Journal of biomolecular screening [J Biomol Screen] 2006 Dec; Vol. 11 (8), pp. 985-95. Date of Electronic Publication: 2006 Nov 07. - Publication Year :
- 2006
-
Abstract
- 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.
- Subjects :
- Animals
Antibodies
Bacteriophages genetics
CHO Cells
Cell Line
Cricetinae
Cricetulus
Enzyme-Linked Immunosorbent Assay methods
Humans
Immunoglobulin G metabolism
Sequence Analysis, Protein
Transfection
Viral Load
Antigens, Surface immunology
Bacteriophages immunology
Immunoglobulin Variable Region immunology
Membrane Proteins pharmacology
Neoplasm Proteins pharmacology
Peptide Library
Subjects
Details
- Language :
- English
- ISSN :
- 1087-0571
- Volume :
- 11
- Issue :
- 8
- Database :
- MEDLINE
- Journal :
- Journal of biomolecular screening
- Publication Type :
- Academic Journal
- Accession number :
- 17092910
- Full Text :
- https://doi.org/10.1177/1087057106293841