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.