Predicting Agitation Stability of Monoclonal Antibodies during Developability Assessment

Area of Science:

• Biopharmaceutical Development
• Protein Aggregation Science
• Antibody Engineering

Background:

• Developability assessment is crucial for selecting antibody drug candidates.
• Predicting agitation-induced aggregation from standard developability parameters remains a challenge.

Purpose of the Study:

• To investigate if key biophysical parameters predict agitation-induced aggregation of monoclonal antibodies (mAbs).
• To identify reliable methods for assessing antibody stability under agitation stress.

Main Methods:

• Generated a benchmark dataset of ten approved mAbs characterized for aggregation upon agitation at an air-liquid interface.
• Utilized flow imaging microscopy and machine learning to analyze aggregate morphology.
• Applied fluorescence, light scattering, chromatography, and drop shape analysis to determine developability and surface parameters.

Main Results:

• Agitation-induced aggregation varied significantly among mAbs, primarily due to antibody identity.
• Standard developability parameters (Tm, Tnr, Tagg, kD, HIC retention, refolding yield) did not correlate with aggregation.
• Surface properties (surface pressure, elastic modulus) showed a good correlation with the concentration of agitated-induced aggregates.

Conclusions:

• Standard developability assays have limitations in predicting mAb interfacial stability.
• Antibody behavior at air-liquid interfaces is critical for aggregation during agitation.
• Surface property analysis offers a more promising approach for predicting aggregation susceptibility.