4. Conclusion
In the present study, based on the previous report for the production of NSP free of FMD vaccine by combination 2D-SECĂ—SEC, we proposed a new platform by replacing SEC in the capturing step with strong anion exchange media in order to compensate low sample loading and achieve higher speed and capacity. Alternatively, mathematical modeling and simulation were applied to predict the retention behavior of active ingredient in vaccine and critical impurities such as rDNA or NSPs. Thus, the results confirmed:
1- Strong anion exchange chromatography in combination with high resolution SEC media such as Sup-200 has acceptable performance for production NSP free FMD vaccine from the perspective of purity and virus recovery, higher than 89.8 % and 77.91% respectively.
2- Capability of mathematical modeling and simulation as a complementary methodology to experimental data will provide a powerful approach to predict retention behavior of large biomolecules such as FMD and rDNA in complex chromatographic phenomena such as IEX for development downstream processing of biotherapeutic such as virus vaccine.
3- Analysis of purified FMDV by SDS-PAGE and HP-SEC confirmed the satisfactory purity for final product.
4- Application of MALDI-TOF mass spectrometry in direct analysis of FMDV particles provide more evidence regarding to purity and structural information of purified FMDV.