Key result
Combined anion-exchange and size exclusion chromatography achieves ~90% residual DNA removal in FMD vaccine purification.
Why the study?
Producing purified virus particles with high quality and quantity for vaccine preparation requires a scalable purification procedure in the downstream step.
A combined 2D-AEC×SEC chromatography method with mathematical modeling improves the downstream purification of foot and mouth disease vaccine, achieving high recovery and virus yield.
Supports FMD vaccine process development in preclinical models; leaves open scale-up, validation, and regulatory translation.
The production of purified virus particles with high quality and quantity for vaccine preparation requires scalable purification procedure in downstream step. A purification scheme based on combined strong anion-exchange and size exclusion chromatography (2D -AEC×SEC) is developed for production non-structural protein (NSP) free foot and mouth diseases (FMD) vaccine and the whole procedure is accomplished with 78 % recovery, 85 % virus yield and more than 90 % of residual DNA (rDNA) is removed from the purified vaccine. Due to use AEC as the first column, the injection volume increases four times compare to previous report. Alternatively, a mathematical modeling and simulation approach based on plate model chromatography are developed and matched with the experimental chromatography data to obtain better perception in predicting retention behavior and saving time in downstream scale-up method development. The analysis of purified virus particles by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), dynamic light scattering (DLS) high performance size exclusion chromatography (HP-SEC), matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS), transmission electron microscopy (TEM) and biological test provide to the best quality of purified FMD virus.
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Ghassempour et al. (2020) studied Foot and Mouth Diseases (FMD) vaccine production. Two-Dimensional Anion Exchange coupling Size Exclusion Chromatography (2D-AEC×SEC) was evaluated on Recovery, virus yield, and residual DNA removal. A combined strong anion-exchange and size exclusion chromatography scheme for FMD vaccine purification achieved 78% recovery, 85% virus yield, and >90% residual DNA removal.
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