Venezuelan equine encephalomyelitis virus: concentration, partial purification, inactivation and immunogenicity

Venezuelan equine encephalomyelitis (VEE) TC-84 vaccinal virus, from 10-1. quantities of infected duck embryo fibroblast cell culture fluids, was isolated by combined continuous-flow centrifugation with isopycnic banding in sucrose. Most of the recovered infectivity and hemagglutinating activity were in a single band at a buoyant density (rho) of 1.2. About 90% of the total input protein (450-520 mg) was removed with the effluent, whereas most of the remaining 10% also banded at a rho of 1.2. Infectivity was inactivated with formalin at a final concentration of 0.05% at 37 degrees C for 24 hr. Formalin-inactivated virus retained its immunogenicity and induced VEE virus-specific antibody in horses and guinea pigs. The horses and those guinea pigs that received equivalent doses of vaccine survived after a challenge of their immunity with virulent VEE virus.