title: The immunogenicity of antibody aggregates in a novel transgenic mouse model.
abstract: OBJECTIVE
  Protein aggregates have been discussed as a potential risk factor related to immunogenicity. Here we developed a novel human IgG transgenic (tg) mouse system expressing a mini-repertoire of human IgG1 antibodies (Abs) for the assessment of immunogenic properties of human mAb preparations.
  
  
  METHODS
  Transgenic mice were generated using germline versions of the human Ig heavy chain γ1 (IgH-γ1), and the human Ig light chain (IgL) κ and λ genes. Only the soluble form of human IgH-γ1 was used to avoid expression of the membrane Ig-H chain and concomitant allelic exclusion of endogenous murine Ig genes. IgG1 aggregates were generated by different stress conditions such as process-related, low pH and exposure to artificial light.
  
  
  RESULTS
  The expression of human Ig proteins induced immunological tolerance to a broad range of human IgG1 molecules in the tg mice. Immunization with IgG1 aggregates demonstrated that soluble oligomers induced by significant light-exposure and carrying neo-epitopes induced a strong immune response in tg mice. In contrast, Ab aggregates alone and monomers with neo-epitopes were not immunogenic.
  
  
  CONCLUSIONS
  This mouse model is able to recognize immunogenic modifications of human IgG1. While the degree of stress-induced aggregation varies for different mAbs, our findings using a particular mAb (mAb1) demonstrate that non-covalently modified aggregates do not break tolerance, contrary to widely held opinion. The immunogenic potential of soluble aggregates of human IgG strongly depends on the presence of neo-epitopes resulting from harsh stress conditions, i.e. extensive exposure to artificial light.
authors: Roche Pharmaceutical Research and Early Development, Pharmaceutical Sciences, Roche Innovation Center, Grenzacherstrasse 124, 4070, Basel, Switzerland.
authors: Juliana
authors: J
authors: Bessa
authors: Sabine
authors: S
authors: Boeckle
authors: Hermann
authors: H
authors: Beck
authors: Thomas
authors: T
authors: Buckel
authors: Sonja
authors: S
authors: Schlicht
authors: Martin
authors: M
authors: Ebeling
authors: Anna
authors: A
authors: Kiialainen
authors: Atanas
authors: A
authors: Koulov
authors: Björn
authors: B
authors: Boll
authors: Thomas
authors: T
authors: Weiser
authors: Thomas
authors: T
authors: Singer
authors: Antonius G
authors: AG
authors: Rolink
authors: Antonio
authors: A
authors: Iglesias
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000235:genetics / Q000276:immunology*; D000917:Antibody Formation; D001483:Base Sequence; D005434:Flow Cytometry; D006801:Humans; D007108:Immune Tolerance; D007074:Immunoglobulin G / Q000235:genetics / Q000276:immunology*; D007143:Immunoglobulin Heavy Chains / Q000235:genetics / Q000276:immunology; D007147:Immunoglobulin Light Chains / Q000235:genetics / Q000276:immunology; D008822:Mice, Transgenic / Q000235:genetics / Q000276:immunology*; D008969:Molecular Sequence Data; D066329:Protein Aggregates / Q000235:genetics / Q000276:immunology*; D013315:Stress, Psychological / Q000276:immunology; D019076:Transgenes
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D066329:Protein Aggregates
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