Simple and efficient ultrafiltration method for purification of rotavirus VP6 oligomeric proteins.

ABSTRACT

Bacterial endotoxins, DNA, live viruses, and viral proteins derived from bacterial and baculovirus (BV) expression vectors employed in recombinant protein production contaminate the final product. Density gradient centrifugation is commonly used to partially purify oligomeric proteins, but impurities from the expression system still remain. We describe a simple and rapid ultrafiltration method for final purification of rotavirus VP6 oligomeric nanotubes and nanospheres. Contamination originating from the BV vector used in VP6 production was undetectable. The method is highly efficient, fast and inexpensive and can be used for a small-scale laboratory purification of VP6 protein to replace technically demanding multi-step chromatographic procedures.