STUDIES ON SOME CHEMICAL PROPERTIES OF ARBOR GROUP VIRUSES
PARTIAL PURIFICATION OF JAPANESE B ENCEPHALITIS VIRUS
1962 Volume 12 Issue 6 Pages 225-231
Details
1962 Volume 12 Issue 6 Pages 225-231
Japanese B encephalitis virus is one of the major members of the group B arbor (arthropod-borne) viruses, and it has a great significance in Japan. A number of studies on Japanese B encephalitis virus have been undertaken since long ago, but many physico-chemical problems still remain unsolved in comparison with some other animal viruses.
This paper reports the experimental results which tried to partially purify Japanese B encephalitis virus particle by means of three biochemical techniques.
Results obtained are as follows:
1) Japanese B encephalitis virus (G1 strain) used was grown in trypsinized hamster-kidney cell cultures. The culture medium consisted of 0.5% lactalbumin hydrolysate, 10% bovine serum, 100u/ml penicillin, and 100γ/ml streptomycin in Hanks' balanced salt solution. The crude virus concentration in culture fluid was about 105.0per 0.02ml in mouse-intracerebral LD50, and it was obtained always four days after the inoculation.
2) Culture fluid harvested from the virus-infected cultures, showing characteristic cellular degeneration, was centrifuged at 3, 000r.p.m./min. for 10 minutes to remove crude cell debrises. The supernatant fluid was loaded onto a column containing ECTEOLA-Cellulose and eluted with 0.07M phosphate buffer solution (pH 7.2). Some supernatant fluid was treated with fluorocarbon. By either procedure 95-100% of the virus was practically recovered, and also more than 90% of the nitrogen contained in the original virus material, as measured by micro-Kjeldahl method, was removed. Appreciable partial purification in terms of protein removal by cholesterol column chromatography was also performed.
3) It was finally concluded from the above results that the procedures of ECTEOLA-Cellulose column chromatography or fluorocarbon deproteinization were the simple and convenient biochemical method which can be successfully applicable to the partial purification of Japanese B encephalitis virus particles.
