Comprehensive, Efficient and Reliable Characterization of Monoclonal Antibody Aggregation by High Throughput and Quantitative Methods

摘要

High molecular weight aggregates of biotherapeutic proteins may be induced by light, temperature fluctuations, pH changes and other stress conditions such as mechanical forces. Aggregates present in these protein products can range from small (dimers) to large particulates (sub-visible or even visible particles). These size variants can be formed during the process of drug development, storage, shipment or delivery. Close monitoring of these size variants is critical because immunogenic responses and differences in pharmacokinetics or potency could arise due to the existence of these species in the drug products. Different analytical techniques are often utilized to fully characterize protein aggregation. Both UV-Vis spectroscopy and size exclusion chromatography (SEC) coupled with UV and light scattering detection play important roles in characterizing monomers, dimers and higher order aggregates. In this presentation UV-Vis spectroscopy is used as a high throughput screening method to assess level of aggregation in different monoclonal antibody (mAb) samples. These samples are further evaluated by SEC method using innovative, high-porosity 2.7μm silica particles and a unique hydrophilic bonding chemistry which could provide exceptional stability with minimal nonspecific interactions. Dimers and higher order aggregates can be quantified through UV detection. Furthermore, combining the resolving power of SEC with light scattering detection would help with detection of higher molecular weight species. This set of methods are suitable for routine aggregate analysis of mAb products.