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Two-Dimensional Anion Exchange coupling Size Exclusion Chromatography Combined with Mathematical Modeling for Downstream Processing of Foot and Mouth Diseases Vaccine
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  • Alireza Ghassempour,
  • Seyed Mohammad Javad Hossienizadeh,
  • Mohsen Bagheri,
  • Mahdi Alizadeh,
  • Masoud Rahimi,
  • Mahmoud Azimi,
  • Morteza Kamalzade,
  • Ali Es-haghi
Alireza Ghassempour
Department of Phytochemistry,
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Seyed Mohammad Javad Hossienizadeh
Shahid Beheshti University
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Mohsen Bagheri
Shahid Beheshti University
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Mahdi Alizadeh
Shahid Beheshti University
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Masoud Rahimi
Shahid Beheshti University
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Mahmoud Azimi
Razi Vaccine & Serum Research Institute
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Morteza Kamalzade
Razi Vaccine & Serum Research Institute
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Ali Es-haghi
Razi Vaccine & Serum Research Institute
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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 -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.

Peer review status:POSTED

11 Sep 2020Submitted to Biotechnology and Bioengineering
11 Sep 2020Assigned to Editor
11 Sep 2020Submission Checks Completed