Plug flow tubular bioreactor, system containing the same and method for production of virus

Abstract

The present invention relates in a first aspect to a plug flow tubular bioreactor having an integral or multi-part tube adapted for cultivation, and, optionally, infection, viral transduction, or transfection of eukaryotic cells. In a further aspect, the present invention relates to a plug flow tubular bioreactor system comprising the plug flow tubular bioreactor according to the present invention. Further, the present invention relates to a method for preparing virus particles, vectors, cells or other molecules including toxic molecules using the plug flow tubular bioreactor or the system according to the present invention. Finally, the present invention relates to the use of a plug flow tubular bioreactor for the preparation of virus particles, vectors including viral vectors, cells including modified cells or other molecules including toxic molecules.

1 . A plug flow tubular bioreactor having an integral or multi-part tube with at least one inlet and at least one outlet whereby the at least one inlet is present at a first end of the tube and the at least one outlet is present at a second end of the tube, wherein the tubular bioreactor is adapted for cultivation, and, optionally, infection, viral transduction, or transfection of eukaryotic cells, and wherein a ratio of a length to a diameter of the tube is at least 5000:1. 
     
     
         2 . A plug flow tubular bioreactor according to  claim 1  further comprising one or more additional inlets and outlets positioned along the length of the tube of said tubular bioreactor for supplying one or more of air, nutrients, and medium. 
     
     
         3 . A plug flow tubular bioreactor according to  claim 1  wherein the tube is at least partly of a hollow fiber. 
     
     
         4 . A plug flow tubular bioreactor according to  claim 1  wherein the tubular bioreactor is configured for single use. 
     
     
         5 . The plug flow tubular bioreactor according to  claim 1  further comprising at least one inlet port for connecting the at least one inlet of the bioreactor to a feed line for introducing cells and, optionally, means for infection, viral transduction, or transfection of said cells, and at least one discharge port for connecting the at least one outlet of the bioreactor with a discharge line. 
     
     
         6 . A plug flow tubular bioreactor system, comprising:
 a plug flow tubular bioreactor according to  claim 1 ;   a module for holding the eukaryotic cells to be cultured and i) infected, virally transduced and or transfected and/or ii) induced or activated for expression of molecules including toxic molecules in the tubular bioreactor;   a module for admixing the eukaryotic cells and i) means for infection, viral transduction, or transfection of the eukaryotic cells and/or ii) means for induction or activation of expression of molecules including toxic molecules; and   at least one pump for allowing plug flow of said mixture through the tubular bioreactor.   
     
     
         7 . The plug flow tubular bioreactor system according to  claim 6  further comprising a control module for control of at least the flow of a) the mixture of cells through the plug flow tubular bioreactor and admixing performed using the module for admixing, and optionally b) the temperature of the bioreactor. 
     
     
         8 . The plug flow tubular bioreactor system according to  claim 6  further comprising means for introducing at least one of air, nutrients, medium, pH-adjusting components, inducers of production into a feed line connected to the tube and/or into the tube. 
     
     
         9 . The plug flow tubular bioreactor system according to  claim 6  wherein the module for holding the cells to be cultured in the tubular bioreactor is a cell bioreactor allowing cultivation of eukaryotic cells. 
     
     
         10 . The plug flow tubular bioreactor system according to  claim 6  further comprising a container containing a virus stock or nucleic acid based compounds. 
     
     
         11 . The plug flow tubular bioreactor system according to  claim 6  further comprising a medium stock reservoir, and/or at least one further pump for introducing medium stock into the plug flow tubular bioreactor. 
     
     
         12 . The plug flow tubular bioreactor system according to  claim 6  further comprising means for controlling at least the pH and/or oxygen concentration of a medium containing the cells and i) the means for infection, viral transduction or transfection or ii) the means allowing induction or activation of expression of molecules including toxic molecules when introduced into the plug flow tubular bioreactor. 
     
     
         13 .- 18 . (canceled) 
     
     
         19 . The plug flow tubular bioreactor of  claim 1  wherein the tube is of silicone-based material. 
     
     
         20 . The plug flow tubular bioreactor system of  claim 6  wherein the plug flow tubular bioreactor is in a coiled configuration. 
     
     
         21 . The plug flow tubular bioreactor system of  claim 6  further comprising one or more sensors determining pH and/or oxygen in the plug flow tubular bioreactor.