Some comments concerning purification of bovine enterovirus.

There are various purification procedures for enteroviruses, depending upon the purpose of the experiment. HeLa S3 cells were grown at 37°C in a Roux bottle for virus multiplication. For the MZ-468 strain of bovine enterovirus, about 11 Roux bottles were sufficient for purification. 10 % polyethylene glycol 6000 was selected for the first virus precipitation step. This purification process gave a recovery of 43 % . A sucrose density gradient (SDG) was successfully formed with a hand -made apparatus, and was fractionated with a hand-made collecting system using a peristalic pump. CsCl bouyant density gradient centrifugation was superior to SDG centrifugation. Even after one cycle of purification with CsCl, a comparatively pure virus preparation was obtained. The purity was checked by electron microscopy and SDS-PAGE analysis of virion proteins. Infectious virion RNA was obtained from the virions which were purified according to this method.