Purification of Japanese encephalitis virus
AssigneeCenter for Disease Control
InventorsMing-Yi Liau, Ruwen Jou, Aih-Jing Chiou
Life of this patent
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Abstract
This invention provides a process for large-scale purification of living Japanese encephalitis virus (JEV) suitable for use in vaccine preparation from a JEV source, including a JEV-infected cell culture and a JEV-infected mouse brain. The process includes the following steps: (a) obtaining a sample from a JEV-infected mouse brain or a JEV-infected cell culture, (b) subjecting the sample to a preliminary separation to remove cell and cell debris from the sample of step (a), (c) concentrating the sample from step (b) by ultra-filtration to remove substances having molecular weight below 100 kDa, and (d) subjecting the sample concentrate to gel filtration to obtain a substantially pure fraction of JEV.
Claims17
An essentially pure live Japanese encephalitis virus preparation which is purified by the process comprising:
- a. obtaining a composition suitable for purification from a Japanese encephalitis virus-infected cell culture;
- b. removing cells and cell debris from said composition by microfiltration;
- c. conducting ultrafiltration directly on the product of step (b);
- d. subjecting the product of step (c) directly to gel filtration to obtain said pure live Japanese encephalitis virus preparation.
A process for large-scale purification of essentially pure live Japanese encephalitis virus preparation for use in vaccine preparation comprising:
- a. obtaining a composition suitable for purification from a Japanese encephalitis virus-infected cell culture or Japanese encephalitis virus-infected mouse brain tissue;
- b. removing cells and cell debris from said composition by microfiltration;
- c. conducting ultrafiltration directly on the product of step (b);
- d. subjecting the product of step (c) directly to gel filtration to obtain said pure live Japanese encephalitis virus preparation.
A method for preparing an essentially pure Japanese encephalitis virus vaccine comprising:
- a. providing an essentially pure live Japanese encephalitis virus preparation which is purified by the process comprising:
- 1. obtaining a composition suitable for purification from a Japanese encephalitis virus-infected cell culture or Japanese encephalitis virus-infected mouse brain tissue;
- 2. removing cells and cell debris from said composition by microfiltration;
- 3. conducting ultrafiltration directly on the product of step (2);
- 4. subjecting the product of step (3) directly to gel filtration to obtain said pure live Japanese encephalitis virus preparation;
- and
- b. inactivating said live Japanese encephalitis virus preparation thereby producing a Japanese encephalitis virus vaccine.
Dependencies are inferred from the claim text.