| 摘要 |
<p>PURPOSE: To obtain a high-quality hepatitis A virus vaccine(HAV) having high qualities of reproducibility, by culturing a HAV, carrying out detergent harvest of the virus, nuclease treatment, concentration, extraction with an organic solvent and purification. CONSTITUTION: A HAV is cultured in a cell sheet of a stationary surface reactor of a large area, a detergent is permeated into the infected cell sheet so as to liberate the HAV from the cell culture product. Then a non-HAV specific nucleic acid is removed from the harvested HAV by nuclease treatment. Then the HAV is concentrated by membrane and dialysis filtration or by capturing by ion exchange, extracted with a solvent such as chloroform-isoamyl alcohol and is purified by ion exchange chromatography and gel filtration chromatography. A HAV having >=95% purity with respect to protein by SD polyacrylamide gel electrophoresis and silver dyeing analysis. The levels of DNA, carbohydrate, RNA and lipid are characterized in terms of vaccine.</p> |
