Purification of Japanese encephalitis virus vaccine by zonal centrifugation

Abstract

A large-scale procedure for purification and concentration of Japanese encephalitis virus vaccine by a continuous-flow isopycnic banding technique in a sucrose density gradient solution, using a K-III zonal centrifuge rotor, is presented. The quality of zonal-purified vaccine was compared with commercial and Japanese National Institutes of Health reference vaccines for antigenicity, immunodiffusion, and allergic encephalitogenicity tests to show its high purity.

Downstream processing of cell culture-derived virus particles

“…The latter is based on the buoyant density of the particle and the rate zonal separation on the particle size and mass. Examples of virus particles that were purified by gradient centrifugation techniques are feline leukemia virus [48], Gibbon ape lymphoma virus [49], hepatitis B virus [50], influenza virus [51], Japanese encephalitis virus [52], mammalian type C virus [53], mouse mammary tumor virus [54], mouse oncorna virus [55], mumps virus [56], murine leukemia virus [57], tick-borne encephalitis virus [58], turkey coronavirus [25], rabies virus [59] and Vaccinia virus [60]. An alternative centrifugation method is the differential centrifugation where pelleting (Epstein-Barr virus [61] and hepatitis A virus [62]) or simple sedimentation onto a sucrose cushion (Moloney murine sarcoma virus [63]) is done once or repeatedly at different centrifugation speeds.…”

Large-scale purification of Japanese encephalitis virus from infected mouse brain for preparation of vaccine

“…Various procedures for separation of brain components from viral suspension have been described, including protamine sulfate treatment (1, 2), fluorocarbon treatment (15), aluminium phosphate gel chromatography (13), ethanol precipitation (11), and sucrose gradient centrifugation (12). In spite of such studies, difficulty in minimizing the amount of host protein still remains in large-scale purification procedures.…”