Two-Dimensional Anion Exchange coupling Size Exclusion Chromatography
Combined with Mathematical Modeling for Downstream Processing of Foot
and Mouth Diseases Vaccine
Abstract
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 -AECĂ—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.