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A Comparison of Four Methods Used to Concentrate Rous Sarcoma Virus from Tissue Culture Fluids

A. Y. Elliott^*,,

Donna Ritzi

^* Department of Urologic Surgery
Department of Microbiology, University of Minnesota Health Sciences, 412 Union Street Southeast, Minneapolis, Minnesota, U.S.A. 55455

Three methods of pelleting, pelleting followed by Pronase treatment, polyethylene glycol (PEG)-Pronase, and diaflo ultrafiltration (diafiltration) were used to concentrate RSV (RAV-1) from tissue culture fluids. Sucrose-gradient fractions containing virus preparations which had been concentrated by diafiltration or pelleting were heavily contaminated with amorphous debris. This debris was not present in similar, gradient-purified preparations that had been concentrated by the PEG-Pronase or pellet-Pronase methods. Maximum recovery of radiolabelled virus particles and virion-associated RNA-dependent DNA polymerase activity was obtained in gradient fractions containing virus concentrates prepared by the pellet-Pronase and PEG-Pronase methods. Although there were slight differences in recovery by these two methods, the advantages of the PEG-Pronase method make it the preferred method, especially when large volumes of tissue culture fluids are used.

Received 3 March 1976; accepted 2 July 1976.