Identification of potent human anti-IL-1RI antagonist antibodies |
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Authors: | Fredericks Zoey L Forte Carla Capuano Irene V Zhou Hongxing Vanden Bos Tim Carter Paul |
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Affiliation: | Department of Antibody Technologies, Amgen Inc., 51 University Street, Seattle, WA 98101-2936, USA. frederiz@amgen.com |
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Abstract: | 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-1R(I) using phage libraries that display single-chain variable fragment (scFv) antibody fragments. Panning libraries against human IL-1R(I) generated 39 unique scFv-phage whose binding to IL-1R(I) was competed by IL-1 ligands. Fifteen of these scFv-phage, identified using IL-1R(I)-binding assays and dissociation rate ranking, were reformatted as scFv-Fc and IgG(4) 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-kappaB nuclear translocation induced by IL-1. Reformatting these clones as IgG(4) molecules increased their inhibition potency by =24-fold. C10 IgG(4) is the most potent antagonist of IL-1alpha (26 nM IC(50)) and IL-1beta (18 nM IC(50)) in the NF-kappaB bioassay, although less potent than IL-1ra ( approximately 0.4 nM IC(50)). C10 is the highest affinity clone for human IL-1R(I) (K(D) approximately 60 nM). Flow cytometry indicates that several lead clones bind cell-surface cynomolgus or murine IL-1R(I), 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. |
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Keywords: | antibody/IL-1/interleukin-1 receptor/phage display/rheumatoid arthritis/scFv |
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