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Interfacial Stress and Container Failure During Freezing of Bulk Protein Solutions Can Be Prevented by Local Heating

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Abstract

Bottles and carboys are used for frozen storage and transport of biopharmaceutical formulations under a wide range of conditions. The quality of freezing and thawing in these systems has been questioned due to the formation of heterogeneous ice structures and deformation of containers. This work shows that during freezing of bulk protein solutions, the liquid at the air–liquid interface freezes first, forming an ice crust and enclosing the liquid phase. As the enclosed liquid freezes, internal pressure rises, pushing the liquid phase through the porous ice crust towards the air interface, leading to interfacial stress and protein aggregation. The aggregation of bovine serum albumin was more intense in the foam-like ice mound that was formed at the top, where bubbles were entrapped. This was characterized experimentally with the assistance of magnetic resonance imaging (MRI). An isothermal cover is proposed to prevent the early freezing of the liquid at the air interface, attenuating substantially interfacial stress to proteins and releasing hydrostatic pressure, preserving the shape and integrity of the containers.

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Acknowledgments

The authors would like to thank Cellon (Luxembourg) for providing the PharmaTainers.

Funding

This work was supported by the Portuguese national funds from Portugal 2020, Lisboa 2020, and European Union through Fundo Europeu do Desenvolvimento Regional (Project 17653, Cryocube).

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Authors and Affiliations

  1. SmartFreeZ, Ed. Inovação II, Incubadora Taguspark, Porto Salvo, Portugal

    Andreia Duarte & Pedro Rego

  2. Instituto Politécnico de Lisboa, Escola Superior de Tecnologia da Saúde de Lisboa (ESTeSL), Lisbon, Portugal

    Aida Ferreira & Paulo Dias

  3. CeFEMA, Instituto Superior Técnico, Universidade de Lisboa, Lisbon, Portugal

    Vítor Geraldes

  4. CQE, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais-1, 1049-001, Lisbon, Portugal

    Miguel A. Rodrigues

Authors
  1. Andreia Duarte
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  2. Pedro Rego
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  3. Aida Ferreira
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  4. Paulo Dias
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  5. Vítor Geraldes
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  6. Miguel A. Rodrigues
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Corresponding author

Correspondence to Miguel A. Rodrigues.

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SmartFreeZ has filed for intellectual property rights on devices related to the science reported in the manuscript.

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Duarte, A., Rego, P., Ferreira, A. et al. Interfacial Stress and Container Failure During Freezing of Bulk Protein Solutions Can Be Prevented by Local Heating. AAPS PharmSciTech 21, 251 (2020). https://doi.org/10.1208/s12249-020-01794-x

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