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A Rapid, Small-Volume Approach to Evaluate Protein Aggregation at Air-Water Interfaces
Caitlin V Wood1, Vladimir I Razinkov2, Wei Qi2
1Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE 19716, USA.
Journal of Pharmaceutical Sciences
|December 3, 2020
Summary
This study presents a fast, small-volume method to assess protein aggregation at air-water interfaces. pH-driven electrostatic interactions significantly impact aggregation, while nonionic surfactants mitigate it for monoclonal antibodies.
Area of Science:
- Biopharmaceutical science
- Protein chemistry
- Physical chemistry
Background:
- Protein aggregation is a critical issue in biopharmaceutical development, affecting drug safety and efficacy.
- Understanding aggregation mechanisms under various stress conditions is essential for formulation development.
- Air-water interfaces (AWI) are known to induce protein aggregation, posing a challenge for manufacturing and storage.
Purpose of the Study:
- To introduce a novel, rapid, small-volume method for evaluating protein aggregation propensity at AWI.
- To investigate the influence of pH and temperature on surface-mediated aggregation of model proteins.
- To assess the impact of nonionic surfactants on mitigating AWI-induced protein aggregation.
Main Methods:
- Utilized a microtensiometer to aerate small protein solution volumes with microbubbles for short durations (≤10 seconds).
- Employed backgrounded membrane imaging to capture and analyze sub-visible particles formed at the AWI.
- Tested aggregation behavior of two model monoclonal antibodies (MAbs) and an alpha-crystallin (aCgn) protein across varying pH and temperature conditions.
Main Results:
- Temperature showed a negligible effect on aggregation under the rapid interface turnover conditions of this technique.
- pH-mediated electrostatic protein-protein interactions were identified as a key driver of particle formation at AWI.
- Nonionic surfactants effectively reduced particle formation in MAb solutions but not in the aCgn solution.
Conclusions:
- The developed microtensiometer method offers a rapid and sample-sparing approach for assessing AWI-induced protein aggregation.
- Electrostatic interactions, influenced by pH, play a more significant role in surface-mediated aggregation than temperature under these conditions.
- Surfactant efficacy in mitigating aggregation is protein-specific, with nonionic surfactants proving beneficial for MAbs but not aCgn.
Keywords:
AdsorptionMonoclonal antibody(s)Physical stabilityProtein aggregationProtein formulation(s)
