Method for reproducible large volume production and purification of Rauscher murine leukemia virus

Abstract

Rauscher murine leukemia virus was produced in roller bottle cultures of chronically infected JLS V9 cells. Virus from this culture fluid was concentrated and purified by two semi isopycnic bandings in sucrose gradients. Virus material obtained from young, nonconfluent cultures (early harvest virus) yielded products characteristically containing endogenous ribonucleic acid dependent deoxyribonucleic acid polymerase with high specific activity (400 to 1,000 pmol of [3H]thymidine 5' triphosphate incorporated per milligram of protein per hour). Fluids obtained from older confluent cultures (late harvest virus) yielded products with endogenous ribonucleic acid dependent deoxyribonucleic acid polymerase with little or no specific activity (200 pmol or less of [3H]thymidine 5' triphosphate incorporated per milligram of protein per hour), but with higher virus particle counts and greater amounts of protein and gs antigen than the early harvest products.