Production and Purification of Large Amounts of Rous Sarcoma Virus

Abstract

Procedures are described for production and purification of large amounts of Rous sarcoma virus. The virus was produced by Rous sarcoma virus-transformed chicken embryo fibroblasts in roller culture which produced up to 6 mg of virus per day per liter of supernatant fluid. Various methods of concentrating virus were evaluated; pelleting yielded the best results in terms of recovery of infectious virus. Purification was achieved by means of successive velocity and equilibrium density centrifugation by using sucrose solutions made in low-salt buffer. A rapid method for the optical density measurement of virus concentration was also developed.