engleski

Purification of measles virus on monolithic columns

Measles virus is used for production of measles vaccine and is found very suitable as a gene vector due to its safety and immunogenicity and also has the ability to specifically kill cancer cells. Most prominent problem with the measles virus as a gene vector or oncolytic virotherapeutic is the large amount of virus needed. Production of measles virus encompasses virus multiplication in live cells followed by formulation of the final product which has to fulfill regulative requirements regarding the proper concentration and purity to ensure safety. Therefore, removal of contaminants i.e. purification which is enabling concentrating of the virus at the same time would be highly beneficial. Traditional purification methods such as ultracentrifugation or ultrafiltration are time consuming and are also not satisfying large scale production needs. In this work purification was based on chromatography using monolith stationary phases which are considered as the fourth– generation chromatography material, featured by very high porosity, high binding capacity and convective based transport which are making them especially well-suited for separation of biomolecules. We tested ion exchangers with small (d =1.35 µm) and large pores (d = 6 µm), at flow rates up to 15 mL/min, using various buffers. Concentration factor up to 30 was achieved but recovery was only up to 25 % although minimal loss in the flow through fraction was found. Possible impact of chromatography on the virus integrity will be investigated in future experiments as well as different monolith chemistries.

measles; monoliths; virus purification

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