title: Role of cosolutes in the aggregation kinetics of monoclonal antibodies.
abstract: We propose a general strategy based on kinetic analysis to investigate how cosolutes affect the aggregation behavior of therapeutic proteins. We apply this approach to study the impact of NaCl and sorbitol on the aggregation kinetics of two monoclonal antibodies, an IgG1 and an IgG2. By using a combination of size exclusion chromatography and light scattering techniques, we study the impact of the cosolutes on the monomer depletion, as well as on the formation of dimers, trimers, and larger aggregates. We analyze these macroscopic effects in the frame of a kinetic model based on Smoluchowski's population balance equations modified to account for nucleation events. By comparing experimental data with model simulations, we discriminate the effect of cosolutes on the elementary steps which contribute to the global aggregation process. In the case of the IgG1, it is found that NaCl accelerates the kinetics of aggregation by promoting specifically aggregation events, while sorbitol delays the kinetics of aggregation by specifically inhibiting protein unfolding. In the case of the IgG2, whose monomer depletion kinetics is limited by dimer formation, NaCl and sorbitol are found respectively to accelerate and inhibit conformational changes and aggregation events to the same extent.
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zurich , Zurich, Switzerland.
authors.forename: Lucrèce
authors.initials: L
authors.lastname: Nicoud
authors.forename: Margaux
authors.initials: M
authors.lastname: Sozo
authors.forename: Paolo
authors.initials: P
authors.lastname: Arosio
authors.forename: Andrew
authors.initials: A
authors.lastname: Yates
authors.forename: Edith
authors.initials: E
authors.lastname: Norrant
authors.forename: Massimo
authors.initials: M
authors.lastname: Morbidelli
mesh_terms: D000911:Antibodies, Monoclonal / Q000737:chemistry*; D002850:Chromatography, Gel; D003198:Computer Simulation; D007074:Immunoglobulin G / Q000737:chemistry*; D007700:Kinetics; D008027:Light; D008956:Models, Chemical; D011487:Protein Conformation; D055503:Protein Multimerization; D055550:Protein Stability; D012542:Scattering, Radiation; D012965:Sodium Chloride / Q000737:chemistry*; D012996:Solutions; D013012:Sorbitol / Q000737:chemistry*; D055672:Static Electricity
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D012996:Solutions; D012965:Sodium Chloride; D013012:Sorbitol