Long-term stability of Vero cell-derived inactivated Japanese encephalitis vaccine prepared using serum-free medium.
We established a method of producing a Vero cell-derived Japanese encephalitis vaccine using serum-free medium, and tested its stability using various stabilizers during the inactivation process and storage at 4 degrees C and 28 degrees C. Similar to previously reported results of cell culture in serum-containing medium, Vero cells were cultured in a serum-free medium multiplied well, and the viral yield was successfully increased to about 10(9)PFU/ml. Following formalin-inactivation and purification via ethanol precipitation and sucrose density ultracentrifugation of the virus solution, the vaccine had the same quality as, and higher immunogenicity, the mouse brain-derived vaccine in current use. Testing of several stabilizers showed that the addition of 0.5% glycine during the virus inactivation process facilitated the maintenance of immunogenicity for a long period of time. Furthermore, the addition of 0.5% glycine and 1.0% sorbitol as vaccine stabilizers after purification led to the maintenance of immunogenicity for 1 year, not dependent on the storage temperature (4 degrees C or 28 degrees C). These results indicate that, in contrast to the current mouse brain-derived vaccine, the Vero cell-derived vaccine can be prepared using serum-free medium containing no animal-derived components, and that the vaccine can be stored at room temperature by adding stabilizers, suggesting the possibility of producing room temperature-stable vaccines.
Department of Research and Development, Research Center for Biologicals, The Kitasato Institute, 6-111 Arai, Kitamoto-shi, Saitama 364-0026, Japan.
Hiroko
H
Toriniwa
Tomoyoshi
T
Komiya
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D018929:Cell Culture Techniques / Q000379:methods*; D002499:Centrifugation, Density Gradient; D005591:Chemical Fractionation; D002522:Chlorocebus aethiops; D016895:Culture Media, Serum-Free; D004355:Drug Stability; D004664:Encephalitis Virus, Japanese / Q000254:growth & development / Q000302:isolation & purification; D005260:Female; D005998:Glycine / Q000494:pharmacology; D022321:Japanese Encephalitis Vaccines / Q000276:immunology*; D051379:Mice; D009500:Neutralization Tests; D013012:Sorbitol / Q000494:pharmacology; D013696:Temperature; D014709:Vero Cells; D038301:Virus Inactivation
D000914:Antibodies, Viral; D016895:Culture Media, Serum-Free; D022321:Japanese Encephalitis Vaccines; D013012:Sorbitol; D005998:Glycine