Skip Navigation

This Article
Right arrow Full Text Freely available
Right arrow FREE Full Text (PDF) Freely available
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in ISI Web of Science
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Search for citing articles in:
ISI Web of Science (7)
Right arrowRequest Permissions
Google Scholar
Right arrow Articles by Fredericks, Z. L.
Right arrow Articles by Carter, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Fredericks, Z. L.
Right arrow Articles by Carter, P.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

Protein Engineering, Design and Selection vol. 17 no. 1 pp. 95-106, 2004
© 2004 Oxford University Press

Identification of potent human anti-IL-1RI antagonist antibodies

Zoey L. Fredericks1,3, Carla Forte1, Irene V. Capuano1, Hongxing Zhou1, Tim Vanden Bos1 and Paul Carter2

1Department of Antibody Technologies, Amgen Inc., 51 University Street, Seattle, WA 98101-2936 and 2Seattle Genetics Inc., 21823 30th Drive S.E., Bothell, WA 98021, USA

3 To whom correspondence should be addressed. e-mail: frederiz{at}amgen.com

Interleukin-1 (IL-1) blockade by IL-1 receptor antagonist benefits some arthritis patients by reducing joint damage. This fact inspired us to develop antagonist human therapeutic antibodies against IL-1RI using phage libraries that display single-chain variable fragment (scFv) antibody fragments. Panning libraries against human IL-1RI generated 39 unique scFv-phage whose binding to IL-1RI was competed by IL-1 ligands. Fifteen of these scFv-phage, identified using IL-1RI-binding assays and dissociation rate ranking, were reformatted as scFv-Fc and IgG4 molecules. The ease of producing antibodies in the scFv-Fc format permitted rapid identification of four lead clones (C10, C13, C14, C15) that inhibit NF-{kappa}B nuclear translocation induced by IL-1. Reformatting these clones as IgG4 molecules increased their inhibition potency by <=24-fold. C10 IgG4 is the most potent antagonist of IL-1{alpha} (26 nM IC50) and IL-1ß (18 nM IC50) in the NF-{kappa}B bioassay, although less potent than IL-1ra (~0.4 nM IC50). C10 is the highest affinity clone for human IL-1RI (KD ~60 nM). Flow cytometry indicates that several lead clones bind cell-surface cynomolgus or murine IL-1RI, characteristics advantageous for preclinical toxicology and efficacy studies. This study demonstrates the utility of scFv-Fc fusion proteins for rapid screening of clones derived from phage libraries to identify antibody leads with therapeutic potential.

Received October 3, 2003; revised October 23, 2003; accepted October 28, 2003 Edited by Greg Winter


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?


This article has been cited by other articles:


Home page
 J. Biol. Chem.Home page
L. Galibert, G. S. Diemer, Z. Liu, R. S. Johnson, J. L. Smith, T. Walzer, M. R. Comeau, C. T. Rauch, M. F. Wolfson, R. A. Sorensen, et al.
Nectin-like Protein 2 Defines a Subset of T-cell Zone Dendritic Cells and Is a Ligand for Class-I-restricted T-cell-associated Molecule
J. Biol. Chem., June 10, 2005; 280(23): 21955 - 21964.
[Abstract] [Full Text] [PDF]


Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.