Development and validation of a molecular size distribution method for polysaccharide vaccines

Area of Science:

• Analytical Chemistry
• Pharmaceutical Sciences
• Biotechnology

Background:

• Accurate molecular size distribution of vaccines is crucial for manufacturing.
• The reference method using TSK G5000PWXL columns shows variability due to high molecular weight compound elution.
• This variability impacts the reliability of vaccine quality control.

Purpose of the Study:

• To develop and validate an alternative method for determining vaccine molecular size distribution.
• To overcome limitations of the reference method, specifically partial elution in the void volume.
• To establish a robust quality control method for polysaccharide-containing vaccines.

Main Methods:

• High-performance size exclusion chromatography (HPSEC) with refractive index detection.
• Utilized a new chromatographic column with a wider separation range.
• Generated a dextran calibration curve for molecular weight cut-off determination.
• Method validation followed International Conference on Harmonisation (ICH) guidelines.

Main Results:

• The new method demonstrated comparable sensitivity to the reference method for detecting product degradation.
• All validation parameters, including precision (intra-assay < 5%, inter-assay < 10%) and recovery (>70%), met ICH requirements.
• Robustness testing confirmed the method's reliability under varied conditions.
• The alternative method exhibited reduced intra- and inter-assay variability compared to the reference.

Conclusions:

• The developed HPSEC method is a suitable quality control tool for vaccine release and stability monitoring.
• This alternative method provides more consistent and reliable molecular size distribution data.
• The findings support the adoption of this improved method for polysaccharide vaccine analysis.