A comparison of four methods used to concentrate Rous sarcoma virus from tissue culture fluids

Abstract

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.