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Purification of Rubella Virus Particles

Orion Laboratories, Helsinki 51 and Department of Virology, University of Helsinki, Helsinki 25, Finland

A procedure for the purification of rubella virus from infected suspensions of BHK 21 cells resulted in preparations containing 1 to 3 x 10¹⁰ p.f.u./ml. In sucrose gradient centrifugation the area of maximal infectivity (buoyant density of 1.175 g./ml.) coincided with peaks in haemagglutinating activity, and in E 260. It also coincided with presence of virus-like particles, and a sharp band visible by naked eye. Specific activities of the order 30 x 10⁶ p.f.u./µg. protein, 10⁶ HAU/µg. protein, and 5 to 10 x 10⁵ p.f.u./HAU were achieved.

Glutaraldehyde-fixed negatively stained rubella virus preparations showed round particles with rough surfaces measuring 50 to 73 nm. (average 61 nm.) in diameter. Unfixed viruses had a greater size variation, 55 to 89 nm. (average 74 nm.) in diameter, apparently due to deformability and fragility of the particles. Spontaneous and deoxycholate-induced breakdown of the particles showed rupture of the envelope but revealed no characteristic inner structure.

Received 25 October 1968; accepted 20 January 1969.