Identification of potent human anti-IL-1RI antagonist antibodies.
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.
Fredericks
Zoey L
ZL
Department of Antibody Technologies, Amgen Inc., 51 University Street, Seattle, WA 98101-2936, USA. frederiz@amgen.com
Forte
Carla
C
Capuano
Irene V
IV
Zhou
Hongxing
H
Vanden Bos
Tim
T
Carter
Paul
P
D021581:Active Transport, Cell Nucleus; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry*; D001667:Binding, Competitive; D019921:Biotinylation; D002462:Cell Membrane / Q000378:metabolism; D002467:Cell Nucleus / Q000378:metabolism; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D005434:Flow Cytometry; D006367:HeLa Cells; D006801:Humans; D007074:Immunoglobulin G / Q000737:chemistry; D007135:Immunoglobulin Variable Region / Q000737:chemistry*; D020128:Inhibitory Concentration 50; D007375:Interleukin-1 / Q000378:metabolism; D007700:Kinetics; D008024:Ligands; D051379:Mice; D008969:Molecular Sequence Data; D019151:Peptide Library; D010802:Phylogeny; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D021381:Protein Transport; D011817:Rabbits; D017472:Receptors, Interleukin-1 / Q000037:antagonists & inhibitors / Q000737:chemistry*; D053573:Receptors, Interleukin-1 Type I; D013997:Time Factors
D000906:Antibodies; D007074:Immunoglobulin G; D007135:Immunoglobulin Variable Region; D007375:Interleukin-1; D008024:Ligands; D019151:Peptide Library; D017472:Receptors, Interleukin-1; D053573:Receptors, Interleukin-1 Type I; D004247:DNA