Size-exclusion chromatography with multi-angle light scattering for elucidating protein aggregation mechanisms

Area of Science:

• Biochemistry
• Analytical Chemistry
• Protein Science

Background:

• Protein aggregation is a critical process in various biological and pharmaceutical contexts.
• Understanding the mechanisms of irreversible protein aggregation is essential for drug development and disease research.

Purpose of the Study:

• To review the application of size exclusion chromatography with inline multi-angle light scattering (SEC-MALS) for protein systems.
• To elucidate mechanistic details of net-irreversible aggregation processes using SEC-MALS.
• To provide guidance on the application, troubleshooting, and data interpretation of SEC-MALS for protein aggregation studies.

Main Methods:

• Size Exclusion Chromatography (SEC) for separating molecules by size.
• Multi-Angle Light Scattering (MALS) for determining molar mass and size distributions.
• Integration of SEC with MALS (SEC-MALS) for comprehensive analysis of protein aggregates.

Main Results:

• SEC-MALS is a suitable technique for interrogating aggregating protein systems.
• Illustrative examples demonstrate the use of SEC-MALS to test mass-action kinetic models of protein aggregation.
• The chapter addresses limitations and potential errors in SEC-MALS data interpretation for aggregation studies.

Conclusions:

• SEC-MALS provides valuable insights into the mechanisms of irreversible protein aggregation.
• Proper application and interpretation of SEC-MALS data are crucial for accurate analysis of protein aggregation.
• This review serves as a guide for researchers utilizing SEC-MALS in protein aggregation studies.