Method for preparing lyophilized hepatitis A attenuated live vaccine using cell factory
A technology of live attenuated vaccines and hepatitis A, which is applied to medical preparations containing active ingredients, antiviral agents, pharmaceutical formulas, etc. It can solve the problems of unsatisfactory effect and long proliferation cycle of hepatitis virus, and achieve easy scale-up Easy to apply, conducive to attachment and growth, and guaranteed sterility
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Examples
Embodiment 1
[0035] The CELL FACTORIES 10 layers and 36 blocks produced by NUNC Company of Denmark were selected, and the preparation process of freeze-dried live attenuated hepatitis A vaccine was prepared by single-layer infection method.
[0036] 1.2BS cell preparation
[0037] Wash the surface of the selected cells with Hank's solution, then digest the cells with 0.25% trypsin, mix the digested cells with MEM culture medium containing 10% calf serum to make a cell suspension, and divide according to an appropriate ratio Seeds (1:5), aseptically added to the cell factory, 200ml / layer, placed at 37°C±0.5°C for 7 days.
[0038] 2. Single layer infection:
[0039] Carefully check the cells to confirm that the cells are monolayer and free of pollution, discard the old solution, wash the cell surface gently with Hank's solution, control it, add the virus seed dilution (0.03MOI), culture at 35°C±0.5°C for 3.5 hours, replenish Add maintenance solution (MEM culture solution containing 2% calf...
Embodiment 2
[0053] The Cell STACK, 10 layers, 36 pieces produced by the American CORNING company was selected, and the preparation process of the freeze-dried hepatitis A live attenuated vaccine was prepared by a single-layer infection method.
[0054] 1.2BS cell preparation
[0055] Wash the surface of the selected cells with Hank's solution, then digest the cells with 0.25% trypsin, mix the digested cells with MEM culture medium containing 10% calf serum to make a cell suspension, and divide according to an appropriate ratio Seeds (1:5), aseptically added to the cell factory, 200ml / layer, placed at 37°C±0.5°C for 7 days.
[0056] 2. Single layer infection:
[0057] Confirm that the cells are monolayer and free of pollution, discard the old solution, gently wash the cell surface with Hank's solution, control it, add virus seed dilution (0.04MOI), culture at 35°C±0.5°C for 3.5 hours, add maintenance solution ( MEM culture fluid containing 2% calf serum) to 200ml / layer, cultured at 35°C±...
Embodiment 3
[0071] Choose CELLFACTORIES produced by NUNC Company, 10 layers and 12 pieces, and Cell STACK produced by American CORNING Company, 40 layers and 6 pieces, and adopt the suspension infection method to prepare the preparation process of freeze-dried hepatitis A live attenuated vaccine.
[0072] 1. Suspension infection
[0073] Select the cell species, discard the culture medium in the cell factory, gently wash the cell surface with Hank's solution, control it, add digestive juice to digest, and when the cell surface is loose, add the mixed suspension of cells and virus species (0.05MOI) , sort according to the appropriate ratio (1:5), place at 35°C±0.5°C for 2.5 hours to absorb, add growth medium (MEM culture solution containing 15% calf serum) to 220ml / layer, place at 35°C±0.5°C for cultivation, The medium was changed after 7 days.
[0074] 2. Replace maintenance fluid
[0075] The liquid in the cell factory was discarded, and 220ml / layer of maintenance solution was added, a...
PUM
Abstract
Description
Claims
Application Information
- R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com