title: Isolation of high-affinity, neutralizing anti-idiotype antibodies by phage and ribosome display for application in immunogenicity and pharmacokinetic analyses.
abstract: Anti-idiotype antibodies against a therapeutic antibody are key reagents for the development of immunogenicity and pharmacokinetic (PK) assays during pre-clinical and clinical development. Here we have used a combination of phage and ribosome display to isolate a panel of monoclonal anti-idiotype antibodies with sub-nanomolar affinity and high specificity to a human anti-IgE monoclonal antibody. Anti-idiotype antibodies were enriched from scFv libraries using phage display, and a biochemical epitope competition assay was used to identify anti-idiotypes which neutralized IgE binding, which was essential for the intended use of the anti-idiotypes as positive controls in neutralizing anti-drug antibody (Nab) assays. The phage display-derived anti-idiotype antibodies were rapidly affinity-matured using a random point mutagenesis approach in ribosome display. Ten anti-idiotype antibodies with improved neutralizing activity relative to the parent antibodies displayed sub-nanomolar affinity for the anti-IgE antibody, representing up to 20-fold improvements in affinity from just two rounds of affinity-based selection. The optimized anti-idiotype antibodies retained the specificity of the parent antibodies, and importantly, were fit for purpose for use in PK and anti-drug antibody (ADA) assays. The approach we describe here for generation of anti-idiotype antibodies to an anti-IgE antibody is generically applicable for the rapid isolation and affinity maturation of anti-idiotype antibodies to any antibody-based drug candidate.
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune Ltd., Milstein Building, Granta Park, Cambridge CB21 6GH, UK. Electronic address: chins@medimmune.com.
authors.forename: Stacey E
authors.initials: SE
authors.lastname: Chin
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune Ltd., Milstein Building, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Franco
authors.initials: F
authors.lastname: Ferraro
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune Ltd., Milstein Building, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Maria
authors.initials: M
authors.lastname: Groves
authors.affiliation: Department of Clinical Pharmacology and DMPK, MedImmune LLC, Mountain View, CA, USA.
authors.forename: Meina
authors.initials: M
authors.lastname: Liang
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune Ltd., Milstein Building, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Tristan J
authors.initials: TJ
authors.lastname: Vaughan
authors.affiliation: Department of Antibody Discovery and Protein Engineering, MedImmune Ltd., Milstein Building, Granta Park, Cambridge CB21 6GH, UK.
authors.forename: Claire L
authors.initials: CL
authors.lastname: Dobson
mesh_terms: D000888:Antibodies, Anti-Idiotypic / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000276:immunology; D057134:Antibodies, Neutralizing / Q000276:immunology*; D000915:Antibody Affinity / Q000276:immunology; D000917:Antibody Formation / Q000276:immunology*; D000918:Antibody Specificity / Q000276:immunology; D001435:Bacteriophages / Q000276:immunology*; D006801:Humans; D007130:Immunoglobulin Idiotypes / Q000276:immunology*; D019151:Peptide Library; D012270:Ribosomes / Q000276:immunology*
chemical_list: D000888:Antibodies, Anti-Idiotypic; D000911:Antibodies, Monoclonal; D057134:Antibodies, Neutralizing; D007130:Immunoglobulin Idiotypes; D019151:Peptide Library; C026408:anti-IgE antibodies
keywords: Affinity maturation; Anti-idiotype; Antibody; Phage display; Ribosome display