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Work package 7

Vaccine production optimization and development of new research tools

This work package is aimed at investigating how to optimize the biotechnological production process of an identified vaccine candidate at small scales. The propagation of the virus candidate in cell culture will be optimized using design of experiment approach (DoE) for the rational determination of critical factors (Eriksson et al. Design of Experiments: Principles and Applications, Umetrics AB, Sweden, 2000). Our previous results show that by optimizing process factors using the DoE approach, virus replication on production culture cells can be greatly improved. This offers new possibilities for productivity increases and reduction of costs in the production of live attenuated vaccines.

If the novel and effective vaccine strains and/or vectors are to be developed downstream processing of virus based vaccine, giving high yields of pure and concentrated virus, is an important issue. The goal of the WP is to design a fast end effective method for the purification and concentration of virus vaccine strains using a monolith stationary phase, which is based on the knowledge of virus biology. The chromatography on monolith stationary phases has been shown to be particularly suited for viruses (Podgornik i sur., J. Chromatogr, 2013) and it will be thus applied for the development of downstream virus purification and concentration procedures (Forčić et al., J. Chromatogr, 2011). The effectiveness of the chromatographic procedures will be evaluated based on the concentration that can be achieved, the time required, the yield of the total virus quantified by RT-PCR and infective virus based on results of virus titrations and on the purity of virus estimated from the reduction of total protein and cellular DNA content. In addition, for detection and quantification of the non-infective virions and virus aggreagates, a new method based on nanoparticles tracking analysis, NanoSight, will be used.

The results will provide information on predictive capacities of the virus production process in the time unit, as well as the efficiency of the production process. The developed procedures would allow a fast transfer of the process into an GMP environment for the production of preclinical and clinical samples, as well as provide sufficient information for the estimation of regular production sustainability.