PRODUCTION AND PURIFICATION OF MILLIGRAM AMOUNTS OF FOOT-AND-MOUTH DISEASE VIRUS FROM BABY HAMSTER KIDNEY CELL CULTURES.

Abstract

A stable line of baby hamster kidney cells for use in the production of, and subsequent purification of, foot-and-mouth disease virus (FMDV) was grown in large quantities on the cylindrical surfaces of 2-liter Baxter bottles. The bottles, in round wire cages, were rotated on a three-tiered roller mill. The cells retained their rapid growth characteristics and susceptibility to FMDV in a tris(hydroxymethyl)aminomethane buffer-containing medium which was especially formulated for large-scale work. This medium, without being changed, sustained cell growth for 6 to 7 days to yield confluent layers containing 500 to 750 million cells per bottle. In small-scale virus-growth experiments, harvested fluids contained about 10(3.8) to 10(8.8) plaque-forming units (PFU) per ml. This corresponded to a yield of 30 to 50 PFU per cell. In production runs with 190 cultures, the infectious fluids usually contained 10(7.9) to 10(9.2) PFU per ml, and the mass of essentially pure virus obtained therefrom ranged from 7 to 17 mg concomitant with cumulative infectivity recoveries of about 20%.