Submicron Aggregation of Chemically Denatured Monoclonal Antibody

Molecular Pharmaceutics
Jacob B RoweYing Wang

Abstract

Isothermal chemical denaturation (ICD) has been widely used to evaluate the conformational stability of therapeutic proteins such as monoclonal antibodies. However, the chemical unfolding pathway and the subsequent aggregation of antibodies are not yet well-understood. In the present work, we conducted a systematic study on an ICD-induced aggregation of a pharmaceutical monoclonal antibody. Using dynamic light scattering, we monitored formation and growth of submicron aggregates in various buffers. Our experiments revealed a nucleation-controlled submicron aggregation of the antibody in the presence of chemical denaturant. After the unfolded protein reached a steady state, we reduced the denaturant concentration by dilution or dialysis to trigger further aggregation after ICD. In this way, we studied the pH effect on aggregation of the stressed protein after removal of denaturant. The ICD-dilution experiment provides a practical means for studying the propensity of unfolded proteins to form aggregates under various formulation conditions. This unique method allows us to control the degree of protein unfolding and the initiation of post-ICD aggregation.

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Citations

Dec 1, 2019·Biotechnology and Bioengineering·Ruben WälchliMassimo Morbidelli
Mar 22, 2020·European Journal of Pharmaceutics and Biopharmaceutics : Official Journal of Arbeitsgemeinschaft Für Pharmazeutische Verfahrenstechnik E.V·Ruben WälchliPaolo Arosio
Aug 30, 2019·Journal of Pharmaceutical Sciences·Yoann Le BasleValérie Sautou
Jun 14, 2021·Biophysical Chemistry·Gulsum MericChristopher J Roberts
Aug 25, 2020·ACS Medicinal Chemistry Letters·Daiki FujimuraHisataka Kobayashi
Oct 27, 2021·Molecular Pharmaceutics·Diana C GomesChristopher J Roberts

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