In this presentation, I will explain the importance of studying the aggregation of partially unfolded antibodies during candidate selection and formulation development. Furthermore, I will present and discuss orthogonal analytical approaches to study this phenomenon.

The propensity of antibodies for non-specific interactions and self-association are some of the most important developability parameters. I will present highlights from ongoing collaborations with research groups at ETH Zürich and University of Cambridge, on the development of new methodologies that can help us to reliably measure and obtain a better understanding of these phenomena. The methodologies include custom-made microfluidics and A4F and they provide important data that complement existing methods.

A key hurdle when developing high concentration biotherapeutics is the manifestation of high viscosity at therapeutic concentrations, which is problematic for both product manufacturing and final administration. Current standard methods, such as capillary rheometry, can be of limited use due to their sample consumption requirements. This presentation explores 3-Dimensional Dynamic Light Scattering optical microrheology as an attractive alternative to current development paradigms while considering its potential applications and practical limitations.

Mutations stimulating hexamerization can potentiate complement activation and clustering of cell surface receptors. Fc-Fc interactions are enhanced selectively at the cell surface and not in solution (controlled oligomerization). Case studies will be discussed with special emphasis on development of assays demonstrating hexamerization potential, functional and biophysical behavior during long-term stability, and impact of formulation.

Stability data are key aspects of drug development and although being relatively simple on one hand they are very time-consuming on the other hand. Achieving robust and reliable long-term stability predictions from accelerated stability studies would be a holy grail of biologics development. In this presentation, I will demonstrate accurate long-term stability predictions of multiple quality attributes for some proteins from short-term accelerated stability studies by using simple kinetic models.

Appropriate selection of conjugation sites and conjugation technologies is now widely accepted as crucial for the success of antibody-drug conjugates (ADCs). Herein, we present ADCs conjugated by different conjugation methods to different conjugation positions being systematically characterized by multiple in vitro assays as well as in vivo pharmacokinetic (PK) analyses in transgenic Tg276 mice.

Subcutaneous administration of therapeutic proteins has gained increasing importance and allows many benefits for patients. However, since application volumes are limited compared to IV, much higher protein concentrations must be achieved, which promote protein-protein interactions and possibly protein aggregation. Recent developments target excipients that prevent this or completely different technical solutions, namely concentrated suspensions instead of solutions. SubQ application should be in mind already in the design phase of drugs.

Polysorbates commonly used as surfactants in biotherapeutic formulations are known to be prone to degradation, mainly by oxidation or enzymatic hydrolysis. Results of a study that examined the oxidation of polysorbate 80 (PS80) and of a monoclonal antibody (mAb) are presented. In particular, the impact of different conditions and factors was evaluated, focusing on different formulation components and on the presence of iron contaminants.

• Root causes for polysorbate oxidation
• Mitigation stratégies against polysorbate oxidation
• Alternative approaches and surfactants

Development of co-formulated protein products is one of the enhanced formulation strategies to obtain a better product aiming to overcome resistance, eliminate co-administration errors, reduce cost in clinics and provide patients with convenience. Potential protein-protein interactions in the co-formulated products, however, cause potential risks in product stability, biological activities, lead to pharmacokinetic interactions, and impact clinical safety. To ensure the safety and efficacy of the co-formulated products, a risk assessment and phase-appropriate approach for protein-protein interaction characterization are recommended, especially on their impact on the critical quality attributes of the protein components during co-formulated protein product development.