Immunogenicity of different stressed IgG monoclonal antibody formulations in immune tolerant transgenic mice
Open Access
- 1 November 2012
- journal article
- Published by Taylor & Francis Ltd in mAbs
- Vol. 4 (6), 740-752
- https://doi.org/10.4161/mabs.22066
Abstract
The presence of protein aggregates in biopharmaceutical formulations is of great concern for safety and efficacy reasons. The aim of this study was to correlate the type and amount of IgG monoclonal antibody aggregates with their immunogenic potential. IgG degradation was obtained by freeze-thawing cycles, pH-shift cycles, heating, shaking and metal-catalyzed oxidation. The size, amount, morphology and type of intermolecular bonds of aggregates, as well as structural changes and epitope integrity were characterized. These formulations were injected in mice transgenic (TG) for human genes for Ig heavy and light chains and their non-transgenic (NTG) counterparts. Anti-drug antibody (ADA) titers were determined by bridging ELISA. Both unstressed IgG and freeze-thawed formulation did not induce measurable ADA levels. A mild antibody response was obtained in a fairly small percentage of mice, when injected with shaken, pH-shifted and heated formulations. The metal-catalyzed oxidized IgG formulation was the most immunogenic one, in both ADA titers and number of responders. The overall titers of NTG responders were significantly higher than the ones produced by TG mice, whereas there was no significant difference between the overall number of TG and NTG responders. This study reinforces the important role of protein aggregates on immunogenicity of therapeutic proteins and provides new insight into the immunogenic potential of different types of IgG aggregates. The results indicate that the quality of the IgG aggregates has more impact on the development of an immune response than their quantity or size.Keywords
This publication has 54 references indexed in Scilit:
- Highly Aggregated Antibody Therapeutics Can Enhance the in Vitro Innate and Late-stage T-cell Immune ResponsesPublished by Elsevier BV ,2012
- Immunogenicity of Therapeutic Proteins: The Use of Animal ModelsPharmaceutical Research, 2011
- Taylor Dispersion Analysis Compared to Dynamic Light Scattering for the Size Analysis of Therapeutic Peptides and Proteins and Their AggregatesPharmaceutical Research, 2011
- Oxidized and Aggregated Recombinant Human Interferon Beta is Immunogenic in Human Interferon Beta Transgenic MicePharmaceutical Research, 2011
- Strategies for the Assessment of Protein Aggregates in Pharmaceutical Biotech Product DevelopmentPharmaceutical Research, 2010
- Aggregated Recombinant Human Interferon Beta Induces Antibodies but No Memory in Immune-Tolerant Transgenic MicePharmaceutical Research, 2010
- Critical Evaluation of Nanoparticle Tracking Analysis (NTA) by NanoSight for the Measurement of Nanoparticles and Protein AggregatesPharmaceutical Research, 2010
- Extrinsic Fluorescent Dyes as Tools for Protein CharacterizationPharmaceutical Research, 2008
- Clinical response to adalimumab: relationship to anti-adalimumab antibodies and serum adalimumab concentrations in rheumatoid arthritisAnnals Of The Rheumatic Diseases, 2007
- Effects of protein aggregates: An immunologic perspectiveThe AAPS Journal, 2006