Protein Aggregation and Performance Optimization Based on Microconformational Changes of Aromatic Hydrophobic Regions

Area of Science:

• Biochemistry
• Protein Science
• Pharmaceutical Development

Background:

• Protein aggregation is a critical issue in biopharmaceutical manufacturing.
• Understanding protein microconformation's role in aggregation is essential.

Purpose of the Study:

• To investigate how microconformational changes in aromatic hydrophobic regions affect protein aggregation.
• To explore the impact of amine and guanidine additives on protein structure and aggregation.

Main Methods:

• Steady-state fluorescence spectroscopy to analyze tryptophan residue microconformation.
• Proteolysis coupled with electrophoresis and molecular dynamics simulations.
• Assessing aggregation degrees of globular proteins under mechanical stress.

Main Results:

• Additives altered aromatic hydrophobic regions into three states: solvent exposure, core burial, or no change.
• Protein aggregation correlated strongly with microconformational changes.
• Aggregation increased with solvent exposure and decreased with core burial of hydrophobic regions.

Conclusions:

• Microconformation of aromatic hydrophobic regions significantly impacts protein aggregation.
• Controlling these microconformations offers a strategy to manage protein aggregation.
• Findings support future preformulation studies for therapeutic proteins.