Process for producing poxviruses and poxvirus compositions
A composition, a poxvirus technology, applied in the preparation of poxviruses, the use in the preparation of medicines, and the field of extracellular enveloped viruses
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[0109] 1. Preparation of a composition comprising IMV and EEV.
[0110] A. Preparation of CEFs.
[0111] 66 SPF eggs were incubated in 2% formaldehyde solution for 60 seconds. After rinsing with 70% ethanol, the eggs were opened, and the embryos were removed and dissected. The resulting tissue was then digested with dispase (Ul / ml) and triple select (Ul / ml) at 36.5°C for 120 minutes.
[0112] The mixture was filtered to remove undigested tissue and CEFs were collected by centrifugation (2300 rpm, 15 minutes).
[0113] B. CEF culture and infection.
[0114] CEFs were incubated at 36.5° C. for 2 days in 55 1 VP-SFM (invitrogen). The cell culture medium was discarded, and poxvirus (0.05 MOI) was added to 55 l of Eagle basal medium (Invitrogen). The infected packaging cells were then incubated for three days.
[0115] C. Poxvirus purification.
[0116] Collect packaging cells and cell culture fluid. Mixture then at Silverson Homogenize in a L4RL4R high-speed homogenizer ...
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