Paolo Arosio1, Simonetta Rima, Massimo Morbidelli. 1. Department of Chemistry and Applied Biosciences, Institute for Chemical and Bioengineering, ETH Zurich, 8093, Zurich, Switzerland.
Abstract
PURPOSE: To identify the aggregation mechanism and the stability characteristics of three different monoclonal antibodies under acidic conditions. METHODS: The aggregation kinetics is analyzed by a combination of light scattering, size exclusion chromatography and fluorescence techniques and the aggregation data are correlated to protein structure, hydrophobicity, charge and antibody subclass. RESULTS: In the investigated conditions, the antibody aggregation follows a mechanism consisting of two-steps: reversible monomer oligomerization followed by irreversible cluster-cluster aggregation. The kinetics of the two steps is differently affected by the operating conditions: mild destabilizing conditions induce formation of oligomers which are stable within weeks, while stronger denaturing conditions promote aggregation of oligomers to larger aggregates which eventually precipitate. For different antibodies significant differences in both oligomerization and growth rates are found, even for antibodies belonging to the same subclass. For all antibodies the aggregate formation is accompanied by a structure re-organization with an increase in the ordered β-sheet structures. At low pH the aggregation propensity of the investigated antibodies does not correlate with antibody subclass, surface net charge and hydrophobicity of the non-native state. CONCLUSIONS: The aggregation mechanism of three antibodies in acidic conditions as well as differences and analogies in their stability behavior has been characterized.
PURPOSE: To identify the aggregation mechanism and the stability characteristics of three different monoclonal antibodies under acidic conditions. METHODS: The aggregation kinetics is analyzed by a combination of light scattering, size exclusion chromatography and fluorescence techniques and the aggregation data are correlated to protein structure, hydrophobicity, charge and antibody subclass. RESULTS: In the investigated conditions, the antibody aggregation follows a mechanism consisting of two-steps: reversible monomer oligomerization followed by irreversible cluster-cluster aggregation. The kinetics of the two steps is differently affected by the operating conditions: mild destabilizing conditions induce formation of oligomers which are stable within weeks, while stronger denaturing conditions promote aggregation of oligomers to larger aggregates which eventually precipitate. For different antibodies significant differences in both oligomerization and growth rates are found, even for antibodies belonging to the same subclass. For all antibodies the aggregate formation is accompanied by a structure re-organization with an increase in the ordered β-sheet structures. At low pH the aggregation propensity of the investigated antibodies does not correlate with antibody subclass, surface net charge and hydrophobicity of the non-native state. CONCLUSIONS: The aggregation mechanism of three antibodies in acidic conditions as well as differences and analogies in their stability behavior has been characterized.
Authors: Rebecca K Brummitt; Douglas P Nesta; Liuquan Chang; Andrew M Kroetsch; Christopher J Roberts Journal: J Pharm Sci Date: 2011-01-06 Impact factor: 3.534
Authors: Anil K Tyagi; Theodore W Randolph; Aichun Dong; Kevin M Maloney; Carl Hitscherich; John F Carpenter Journal: J Pharm Sci Date: 2009-01 Impact factor: 3.534
Authors: John F Carpenter; Theodore W Randolph; Wim Jiskoot; Daan J A Crommelin; C Russell Middaugh; Gerhard Winter Journal: J Pharm Sci Date: 2010-05 Impact factor: 3.534
Authors: Christopher J Brown; Daniel W Woodall; Tarick J El-Baba; David E Clemmer Journal: J Am Soc Mass Spectrom Date: 2019-07-30 Impact factor: 3.109
Authors: Srivalli N Telikepalli; Ozan S Kumru; Cavan Kalonia; Reza Esfandiary; Sangeeta B Joshi; C Russell Middaugh; David B Volkin Journal: J Pharm Sci Date: 2014-01-22 Impact factor: 3.534
Authors: Yingda Xu; Dongdong Wang; Bruce Mason; Tony Rossomando; Ning Li; Dingjiang Liu; Jason K Cheung; Wei Xu; Smita Raghava; Amit Katiyar; Christine Nowak; Tao Xiang; Diane D Dong; Joanne Sun; Alain Beck; Hongcheng Liu Journal: MAbs Date: 2018-12-17 Impact factor: 5.857
Authors: Maria Monica Castellanos; Jai A Pathak; William Leach; Steven M Bishop; Ralph H Colby Journal: Biophys J Date: 2014-07-15 Impact factor: 4.033