A General Small-Angle X-ray Scattering-Based Screening Protocol for Studying Physical Stability of Protein...

Area of Science:

• Biophysics
• Pharmaceutical Sciences
• Materials Science

Background:

• Selecting stable protein drug formulations is crucial for efficacy and preventing degradation or aggregation.
• Current methods for formulation screening, while useful, may have limitations in sensitivity or compatibility.

Purpose of the Study:

• To design and evaluate a general workflow for screening protein formulations using Small-Angle X-ray Scattering (SAXS).
• To assess SAXS as a complementary technique to established methods like Dynamic Light Scattering (DLS) for high-throughput analysis.

Main Methods:

• Developed an automated SAXS pipeline incorporating sample handling, temperature control, and rapid data analysis.
• Utilized SAXS in conjunction with turbidity analysis, Dynamic Light Scattering (DLS), and SDS-PAGE for comprehensive parameter acquisition.
• Tested the workflow with model proteins and biopharmaceuticals across various buffer, excipient, and concentration conditions.

Main Results:

• SAXS provides a more sensitive measure of protein particle interactions, including aggregation and repulsion, compared to DLS.
• The SAXS workflow demonstrated compatibility with a wider range of formulation components and protein concentrations.
• In situ SAXS proved to be a sensitive indicator of long-term protein stability.

Conclusions:

• The developed SAXS workflow enables high-throughput screening of protein formulations.
• SAXS is a valuable, complementary technique to DLS for biopharmaceutical research and industry.
• This approach can be integrated into existing physicochemical analysis pipelines for enhanced protein stability assessment.