There is a growing industry and regulatory need to detect host cell proteins (HCPs) in the production of protein biopharmaceuticals, as certain HCPs can impact product stability, safely and efficacy, even at low levels. We developed an HCP profiling workflow using an Evosep ONE LC system coupled to an Orbitrap Fusion Lumos mass spectrometer, which led to HCP detection and quantification in a significantly shorter turnaround time. The workflow can be readily implemented and adapted for different purposes to guide biopharmaceutical process development and enable better risk assessment of HCPs in drug substances.

Compendial methods to analyze subvisible particles in biologics often encounter difficulties when applied to proteinaceous drugs. This presentation discusses how to combine AI, computational statistics and high-throughput microscopy to characterize particles (e.g., protein aggregates) in biologics. By analyzing morphological and textural features particles in images, this method delivers quantitative, actionable information for formulation development, container qualification, and fill-finish quality control. Four case studies are discussed.

The therapeutic proteins are manufactured through upstream/downstream process, fill/finish, and delivered to patients. Impurities in therapeutic protein drugs can originate from raw material, bioprocess, product itself, or dosing regimen. This presentation will highlight potential impurities of therapeutic proteins and share the best practice of analytical procedures and characterization technologies to monitor and control impurities. The impurity comments/questions from health authority after IND/IMPD is submitted will be exemplified as case studies.

Virus Like Particles can be used to economically predict viral clearance outcomes during process development and characterization in a BSL-1 setting. This presentation will first review the physicochemical characteristics of a non-infectious Minute Virus of Mice particle and a CHO-endogenous Retrovirus Like Particle. Then, application data will be shown from a series of comparative spiking studies including an AEX DOE study, a HTS resin selection study, and an AAV process study.

In this presentation, we will discuss an approach to perform drug product process development studies to de-risk DP manufacture at commercial manufacturing site. We also discuss a QbD approach for technology transfer.

Real-time analytical techniques provide an opportunity for more effective monitoring and control of protein aggregation in bioprocessing as well as a better understanding of the mechanisms of protein aggregation. The objective of this presentation is to share these opportunities as well as challenges in the implementation of novel technologies that can aid in the detection of protein aggregates in real-time.

Variability throughout bioprocess manufacturing is magnified by impurities introduced by the raw materials and medium mixing protocols. Analytical tools such as the Raman spectrophotometry and focused beam reflectance measurement (FBRM) will allow us to capture the intricate media chemistry that occurs due to the changes introduced by medium raw material impurity or mixing parameters. In this presentation, we will share our strategy and case study on using analytical tools in the characterization of medium preparation.

The aggregation of biotherapeutics is an undesirable phenomenon for various reasons, including reduced efficacy as well as immunogenicity. Nevertheless, the aggregation process is often difficult to understand and more importantly, to predict for newly developed molecules. Here we present a method for studying the surface-mediated stress that leads to aggregation, in a very controlled manner. The method was applied to a library of antibodies to select the best candidates.

Polysorbate 80 (PS80), a nonionic surfacatant used in pharmaceutical formulation, is known to be incompatible with m-cresol, an antimicrobial agent. This incompatibility results in increased turbidity caused by unsteady micelle aggregation or coalescence that can progress over weeks. PS80/m-cresol solution instability is influenced by storage temperature, ionic strength and component concentration. We use small-angle neutron scattering (SANS) to reveal the cause of aggregation, the coalescence mechanism and aggregate structure. From our SANS results, we find that PS80/m-cresol solution phase instability progresses through multiple growth stages that can be explained using a single kinetic model.

Free fatty acid particles associated with enzymatic polysorbate degradation in liquid drug products is of increasing concern to the biopharmaceutical industry. There are currently several gaps in the industry's understanding of how to mitigate this issue. The talk will focus on formulation related considerations and risks associated with polysorbate degradation as well as strategies on how to manage polysorbate degradation and reduce particle risk.