Systems and Methods for Virus Propagation in Cell Cultures for Vaccine Manufacture

摘要

The present invention provides a closed system to propagate virus-infected cells without the effect of shear force, while providing quicker access to nutrients than is available conventionally. This system design allows for a high density of infected cell growth to increase the virus yields and to maintain homogeneity of the contents of the main container. The system further provides a nuclease to degrade the cellular DNA prior for purification of the virus or viral components. As the system is designed for maximum containment at low risk, the live virus can be a hazardous virus such as a Bio-safety Level 3 (BSL 3), BSL 4 or BSL5 virus.

主权项

1. A method of producing bulk quantities of inactivated virus comprising:
providing a closed-loop cell culture system that includes:
one or more incubation vessels each of which contains a matrix material for supporting a population of adherent cells;a media tank containing cell culture media connected to the one or more incubation vessels, wherein the cell culture media flows from the media tank to the one or more incubation vessels and back to the media tank; anda valve downstream of the one or more incubation vessels that, when activated, closes a connection to the media tank and opens a connection to a column containing an affinity matrix, wherein the column possesses an input end connected to the one or more incubation vessels and an output end connected to a collection vessel; contacting the population of adherent cells to the matrix material contained within the one or more incubation vessels; flowing the cell culture medium within the closed-loop cell culture system for a first period of time; contacting the population of adherent cells with live virus creating virus infected cells; culturing the virus-infected cells for a second period of time to generate replicated virus; adding an agent to the virus-infected cells after the second period of time to promote isolation of replicated virus from the virus-infected cells; activating the valve such that replicated virus flows to the column under conditions that promote the selective retention of replicated virus by the affinity matrix; eluting the replicated virus from the affinity matrix and collecting virus in the collection vessel; and inactivating the eluted virus.