Stresses, Stabilization, and Recent Insights in Freezing of Biologics

Freeze-thawing is a critical unit operation during manufacturing of biopharmaceuticals. A significant fraction of biologics drug substances are stored frozen. While the number of frozen drug products are fewer than drug substances, a frozen drug product provides a path forward in early stages of biologics development. There is increasing diversity in the modalities that are categorized as biologics and includes cells, DNA/RNA-based therapeutics, proteins, vaccines, and viruses. Several detailed commentaries are available on the fundamental characteristics of freezing, the resulting stresses, and their consequences on the stability of many modalities, chiefly, proteins. The present chapter provides an overview of the fundamental aspects of freezing and the consequences of the developing stresses on protein stability. The chapter also discusses recent advances in experimental and modeling approaches to investigate freezing-related processes and induced damage. Lastly, the commentary summarizes critical formulation and process considerations (best practices) in the development of biologics drug products (mAbs and beyond), along with the future opportunities that exist in bridging the knowledge gaps in freezing and stabilization of proteins.