A Rapid, Small-Volume Approach to Evaluate Protein Aggregation at Air-Water Interfaces

J Pharm Sci. 2021 Mar;110(3):1083-1092. doi: 10.1016/j.xphs.2020.11.024. Epub 2020 Nov 30.

Abstract

Non-native protein aggregation is a common concern for biopharmaceuticals. A given protein may aggregate through a variety of mechanisms that depend on solution and physico-chemical stress conditions. A thorough evaluation of aggregation behavior for a protein under all conditions of interest is necessary to ensure drug safety and efficacy. This work introduces a rapid, small-volume approach to evaluate protein aggregation propensity upon exposure to air-water interfaces (AWI). A microtensiometer apparatus is used to aerate a small volume of a protein solution with microbubbles for short periods of time (≤10 s). Sub-visible particles that form are captured and analyzed using backgrounded membrane imaging. This allows one to capture all particles in the solution while being sample sparing. The surface-mediated aggregation of two model monoclonal antibodies (MAbs) and a globular protein (aCgn) was tested as a function of pH and temperature. Temperature had a negligible effect under the rapid interface turnover time scales with this technique. Electrostatic protein-protein interactions, mediated by pH changes, were more influential for particle formation via AWI. Nonionic surfactants substantially reduced particle formation for all MAb solutions, but not aCgn. The results are contrasted with expectations when exposing samples to much larger air-water interfacial stress.

Keywords: Adsorption; Monoclonal antibody(s); Physical stability; Protein aggregation; Protein formulation(s).

Publication types

  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Antibodies, Monoclonal
  • Protein Aggregates*
  • Water*

Substances

  • Antibodies, Monoclonal
  • Protein Aggregates
  • Water