A comparison of four methods used to concentrate Rous sarcoma virus from tissue culture fluids.
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.
D001358:Avian Sarcoma Viruses / Q000201:enzymology / Q000302:isolation & purification*; D002499:Centrifugation, Density Gradient; D003470:Culture Media; D011092:Polyethylene Glycols; D011402:Pronase; D012194:RNA-Directed DNA Polymerase / Q000378:metabolism; D014462:Ultrafiltration; D014776:Virus Cultivation / Q000379:methods*
D003470:Culture Media; D011092:Polyethylene Glycols; D012194:RNA-Directed DNA Polymerase; D011402:Pronase