Real-Time Assessment of the Size Changes of Individual Sub-Visible Protein Particles under Buffer Variations: A...

Area of Science:

• Biopharmaceutical Manufacturing
• Protein Aggregation Studies
• Advanced Drug Delivery Systems

Background:

• Protein particles in biological drugs can reduce efficacy and cause adverse effects.
• Controlling protein particle formation in biopharmaceutical manufacturing remains a challenge.
• Novel biologic formats may exhibit increased susceptibility to aggregation.

Purpose of the Study:

• To introduce a microfluidic method for real-time analysis of individual sub-visible protein particles.
• To investigate the impact of modulating intermolecular forces on protein particle behavior.
• To compare the biophysical behavior of protein particles with monomeric protein.

Main Methods:

• Utilized a microfluidic approach for real-time particle analysis.
• Manipulated buffer pH and urea concentration to alter intermolecular forces.
• Observed particle size changes and compared particle behavior to monomeric protein.

Main Results:

• Protein particles exhibited reversible swelling and shrinkage up to 50% with changes in buffer conditions.
• Demonstrated that altered intermolecular distances influence particle size.
• Identified a discrepancy between the biophysical behavior of protein particles and monomeric protein.
• Highlighted limitations of current biophysical methods in predicting particle formation.

Conclusions:

• Studying individual protein particles provides crucial insights into their behavior.
• Particle swelling may indicate manufacturing deviations, impacting drug quality.
• Understanding particle dynamics is essential for robust biopharmaceutical formulation development.