Trehalose Limits Fragment Antibody Aggregation and Influences Charge Variant Formation in Spray-Dried Formulations at...

Area of Science:

• Pharmaceutical Sciences
• Biotechnology
• Materials Science

Background:

• Hot-melt extrusion (HME) is a manufacturing process for sustained-release drug delivery systems, utilizing heat and shear.
• Protein stability in solid-state formulations at elevated temperatures is critical for successful HME of protein-loaded products.

Purpose of the Study:

• To investigate the solid-state stability of a model protein (Fab2) in formulations with trehalose at elevated temperatures.
• To understand the protective mechanism of trehalose against protein degradation and aggregation during thermal processing.

Main Methods:

• Spray-drying of formulations with varying trehalose concentrations.
• Exposure of formulations to temperatures between 90-120 °C.
• Assessment of protein aggregation and chemical degradation rates.
• Analysis of protein secondary structure using circular dichroism (CD).
• Evaluation of particle morphology using scanning electron microscopy (SEM).

Main Results:

• Trehalose demonstrated a concentration-dependent stabilization effect on Fab2, limiting degradation and aggregation.
• The stabilizing effect of trehalose plateaued at a trehalose-to-protein mass ratio of approximately 1.
• CD and SEM analyses provided insights into the mechanism of trehalose-mediated protein stabilization in the solid state.

Conclusions:

• Trehalose effectively stabilizes fragment antibodies in solid-state formulations against thermal degradation.
• The findings support the feasibility of using high-temperature processes like hot-melt extrusion for manufacturing protein-loaded PLGA rods with minimal protein damage.
• Trehalose acts as a crucial excipient for enhancing protein stability during solid-state thermal processing.