Automation of Polysaccharide Quantification: A Rapid High-Throughput Assay Enabled by Liquid Handling Technology

Area of Science:

• Analytical Chemistry
• Biochemistry
• Vaccine Development

Background:

• Traditional anthrone assays are common for polysaccharide quantitation but are labor-intensive and unsuitable for high-throughput needs.
• The corrosive nature of the anthrone-sulfuric acid reagent poses challenges for automation.
• Accurate polysaccharide quantification is crucial for vaccine development and quality control.

Purpose of the Study:

• To develop and validate a fully automated, corrosion-mitigated anthrone assay using a liquid handling system (LHS).
• To assess the accuracy, precision, and linearity of the automated assay compared to the manual method.
• To validate the automated assay according to ICH Q2 (R2) guidelines for reliable polysaccharide quantitation.

Main Methods:

• Development of an automated anthrone assay integrated with a liquid handling system (LHS).
• Quantification of 23 pneumococcal (PNU) polysaccharide serotypes using both manual and automated anthrone assays.
• Validation of the automated LHS method according to ICH Q2 (R2) guidelines, assessing accuracy, precision, and linearity.

Main Results:

• The automated anthrone LHS method achieved accuracy between 81-115% and precision with a coefficient of variation <7.0%.
• Linearity was demonstrated in the 30-650 µg/mL range with R² ≥ 0.993.
• The automated assay processes eight samples per hour, supports overnight batch operation, and automates all steps from pipetting to data export.

Conclusions:

• The developed automated anthrone assay is the first fully unattended and corrosion-mitigated method validated under ICH Q2 (R2) for polysaccharide quantitation.
• This automated LHS method provides an efficient, accurate, and precise alternative to manual anthrone assays for high-throughput analysis.
• The system facilitates overnight batch processing, significantly enhancing workflow efficiency in vaccine component analysis.