title: Functional analysis of the anti-adalimumab response using patient-derived monoclonal antibodies.
abstract: The production of antibodies to adalimumab in autoimmune patients treated with adalimumab is shown to diminish treatment efficacy. We previously showed that these antibodies are almost exclusively neutralizing, indicating a restricted response. Here, we investigated the characteristics of a panel of patient-derived monoclonal antibodies for binding to adalimumab. Single B-cells were isolated from two patients, cultured, and screened for adalimumab specificity. Analysis of variable region sequences of 16 clones suggests that the immune response against adalimumab is broad, involving multiple B-cell clones each using different combinations of V(D)J segments. A strong bias for replacement mutations in the complementarity determining regions was found, indicating an antigen-driven response. We recombinantly expressed 11 different monoclonal antibodies and investigated their affinity and specificity. All clones except one are of high affinity (Kd between 0.6 and 233 pm) and compete with TNF as well as each other for binding to adalimumab. However, binding to a panel of single-point mutants of adalimumab indicates markedly different fine specificities that also result in a differential tendency of each clone to form dimeric and multimeric immune complexes. We conclude that although all anti-adalimumab antibodies compete for binding to TNF, the response is clonally diverse and involves multiple epitopes on adalimumab. These results are important for understanding the relationship between self and non-self or idiotypic determinants on therapeutic antibodies and their potential immunogenicity.
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Pauline A
authors.initials: PA
authors.lastname: van Schouwenburg
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Simone
authors.initials: S
authors.lastname: Kruithof
authors.affiliation: Global Drug Discovery, Global Biologics, Bayer HealthCare AG, 51061 Cologne, Germany, and.
authors.forename: Christian
authors.initials: C
authors.lastname: Votsmeier
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Karin
authors.initials: K
authors.lastname: van Schie
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Margreet H
authors.initials: MH
authors.lastname: Hart
authors.affiliation: Genmab B.V., 3584 BV Utrecht, The Netherlands.
authors.forename: Rob N
authors.initials: RN
authors.lastname: de Jong
authors.affiliation: Genmab B.V., 3584 BV Utrecht, The Netherlands.
authors.forename: Esther E L
authors.initials: EE
authors.lastname: van Buren
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Marieke
authors.initials: M
authors.lastname: van Ham
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Lucien
authors.initials: L
authors.lastname: Aarden
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands, the Jan van Breemen Research Institute/Reade, 1056 AB Amsterdam, The Netherlands.
authors.forename: Gertjan
authors.initials: G
authors.lastname: Wolbink
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Diana
authors.initials: D
authors.lastname: Wouters
authors.affiliation: From the Department of Immunopathology, Sanquin Research and Landsteiner Laboratory Academic Medical Centre, 1105 AZ Amsterdam, The Netherlands, t.rispens@sanquin.nl.
authors.forename: Theo
authors.initials: T
authors.lastname: Rispens
mesh_terms: D000068879:Adalimumab; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology*; D061067:Antibodies, Monoclonal, Humanized / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D057134:Antibodies, Neutralizing / Q000235:genetics / Q000276:immunology; D000918:Antibody Specificity; D057809:HEK293 Cells; D006801:Humans; D007143:Immunoglobulin Heavy Chains / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000276:immunology; D007135:Immunoglobulin Variable Region / Q000276:immunology; D008969:Molecular Sequence Data; D017354:Point Mutation
chemical_list: D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D057134:Antibodies, Neutralizing; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D000068879:Adalimumab
keywords: Adalimumab; Antibody Engineering; Autoimmune Disease; Biotechnology; Humoral Response; Immunogenicity; Monoclonal Antibody; Tumor Necrosis Factor (TNF)