Impact of protein aggregation on the immunogenicity of a human monoclonal antibody following pulmonary administration in mice
Sohaib Mahri1, Céline Cassiers 1, Gracin Sandra2, Francine Uwambayinema5, François Huaux5, Mariam Ibrahim3, Sian Piper4, Antonio Llinas2, Markus Fridén2,6, Rita Vanbever1
1 Université catholique de Louvain (UCLouvain), Louvain Drug Research Institute (LDRI), Advanced Drug Delivery & Biomaterials (ADBB), Brussels 1200, Belgium.
2 AstraZeneca; Pharmaceutical Technology & Development; Inhalation Product Development; Gothenburg, Sweden
3 AstraZeneca; R&D Biopharmaceuticals; Biopharmaceuticals Development; Gaithersburg, US
4 AstraZeneca; R&D Biopharmaceuticals; Bioscience Asthma, Early R&I; Cambridge, UK
5 UCLouvain, Institut de Recherche Expérimentale et Clinique, Louvain Centre for Toxicology and Applied Pharmacology, Brussels, Belgium
6 Uppsala University, Department of Pharmacy, Uppsala, Sweden
Summary
Spray drying is commonly used to produce dry powder formulations for inhalation. However, this process subjects proteins to stresses likely resulting in the formation of protein aggregates. These aggregates can cause a loss of activity and an increased risk of immunogenicity. The impact of aggregates on the immunogenicity of therapeutic proteins delivered via the pulmonary route is not well understood. Therefore, our study aimed to specifically assess the effect of different levels of aggregates on the immunogenicity of the human monoclonal antibody NIP228 after repeated intratracheal administration in mice. NIP228 was labelled with NHS-rhodamine and then aggregate-free rhod-NIP228 solution (No_agg) was spray-dried both with and without 1% polysorbate 80 surfactant, to generate powders with high and low levels of NIP228 aggregates (referred to as Hi_agg and Lo_agg, respectively). Following repeated intratracheal administration of the No_agg, Lo_agg, or Hi_agg in mice, anti-NIP228 antibody (ADA) levels in both sera and bronchoalveolar lavage fluids (BAL) were measured. The incorporation of PS80 significantly reduced the aggregate formation associated with spray drying and increased FPF < 5 μm (78% for Hi_agg vs 87% for Lo_agg ). Interestingly, the level of aggregates did not significantly impact the magnitude of the anti-drug antibody response in sera or BAL and the pulmonary route induced a stronger immune response compared to subcutaneous administration.