title: Use of In Vitro Assays to Assess Immunogenicity Risk of Antibody-Based Biotherapeutics.
abstract: An In Vitro Comparative Immunogenicity Assessment (IVCIA) assay was evaluated as a tool for predicting the potential relative immunogenicity of biotherapeutic attributes. Peripheral blood mononuclear cells from up to 50 healthy naïve human donors were monitored up to 8 days for T-cell proliferation, the number of IL-2 or IFN-γ secreting cells, and the concentration of a panel of secreted cytokines. The response in the assay to 10 monoclonal antibodies was found to be in agreement with the clinical immunogenicity, suggesting that the assay might be applied to immunogenicity risk assessment of antibody biotherapeutic attributes. However, the response in the assay is a measure of T-cell functional activity and the alignment with clinical immunogenicity depends on several other factors. The assay was sensitive to sequence variants and could differentiate single point mutations of the same biotherapeutic. Nine mAbs that were highly aggregated by stirring induced a higher response in the assay than the original mAbs before stirring stress, in a manner that did not match the relative T-cell response of the original mAbs. In contrast, mAbs that were glycated by different sugars (galactose, glucose, and mannose) showed little to no increase in response in the assay above the response to the original mAbs before glycation treatment. The assay was also used successfully to assess similarity between multiple lots of the same mAb, both from the same manufacturer and from different manufacturers (biosimilars). A strategy for using the IVCIA assay for immunogenicity risk assessment during the entire lifespan development of biopharmaceuticals is proposed.
authors: Department of Attribute Sciences, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Marisa K
authors: MK
authors: Joubert
authors: Department of Clinical Immunology, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Meghana
authors: M
authors: Deshpande
authors: Department of Attribute Sciences, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Jane
authors: J
authors: Yang
authors: Antitope Limited, Babraham Research Campus, Cambridge, United Kingdom.
authors: Helen
authors: H
authors: Reynolds
authors: Antitope Limited, Babraham Research Campus, Cambridge, United Kingdom.
authors: Christine
authors: C
authors: Bryson
authors: Antitope Limited, Babraham Research Campus, Cambridge, United Kingdom.
authors: Mark
authors: M
authors: Fogg
authors: Antitope Limited, Babraham Research Campus, Cambridge, United Kingdom.
authors: Matthew P
authors: MP
authors: Baker
authors: Department of Clinical Immunology, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Jonathan
authors: J
authors: Herskovitz
authors: Department of Clinical Immunology, Amgen Inc., Seattle, Washington, United States of America.
authors: Theresa J
authors: TJ
authors: Goletz
authors: Department of Medical Sciences, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Lei
authors: L
authors: Zhou
authors: Department of Clinical Immunology, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Michael
authors: M
authors: Moxness
authors: Department of Attribute Sciences, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Gregory C
authors: GC
authors: Flynn
authors: Department of Attribute Sciences, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Linda O
authors: LO
authors: Narhi
authors: Department of Clinical Immunology, Amgen Inc., Thousand Oaks, California, United States of America.
authors: Vibha
authors: V
authors: Jawa
mesh_terms: D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology* / Q000378:metabolism; D059451:Biosimilar Pharmaceuticals; D049109:Cell Proliferation; D002478:Cells, Cultured; D016207:Cytokines / Q000032:analysis; D004797:Enzyme-Linked Immunosorbent Assay; D058501:Enzyme-Linked Immunospot Assay; D006031:Glycosylation; D006801:Humans; D007371:Interferon-gamma / Q000032:analysis; D007376:Interleukin-2 / Q000032:analysis; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D008213:Lymphocyte Activation; D017354:Point Mutation; D018570:Risk Assessment; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D059451:Biosimilar Pharmaceuticals; D016207:Cytokines; D007376:Interleukin-2; D007371:Interferon-gamma