Single use centrifuge system for highly concentrated and/or turbid feeds

Abstract

A method and apparatus for cell harvest of production scale quantities of cell cultures using single use components includes a flexible membrane mounted on a rigid frame and is supported within a multiple use rigid centrifuge bowl, such single use components including a core with an increased diameter and an internal truncated cone shape in order to permit the system to maintain a sufficiently high angular velocity to create a settling velocity suited to efficiently processing highly concentrated cell culture streams. Features which minimize feed turbidity, and others which permit the continuous or semi-continuous discharge of cell concentrate, increase the overall production rate over the rate which can be achieved using current intermittent processing methods for large cell culture volumes. Injection of a diluent during the cell concentrate removal process permits more complete removal of viscous cell concentrates.

What is claimed is: 
     
         1 . Apparatus comprising:
 a single use structure for a centrifuge system configured to separate cells in suspension in a cell culture batch, comprising:   a rotatable upper flange,   a feed tube, wherein the feed tube is configured to be rotationally fixed during cell suspension processing,
 wherein the feed tube extends along a vertical axis, 
   a centrate discharge passage, wherein the discharge passage extends along the vertical axis,
 wherein the discharge passage comprises a discharge tube that is fluidly connected with a centrate outlet, 
   a rotatable core,
 wherein the core is disposed coaxially with respect to the feed tube, 
 wherein the core is configured to receive cell suspension fed through the feed tube, 
 wherein the upper flange is configured to rotate during cell suspension processing,
 wherein during upper flange rotation, the core rotates relative to the feed tube and the discharge tube, 
 
 wherein the core includes a core interior,
 wherein the feed tube axially extends into the core interior, 
 wherein the core interior includes an upper portion and a lower portion, 
 
   a seal arrangement disposed annularly about the feed tube,
 wherein the seal arrangement provides abutting engagement between the relatively rotationally movable portions of the single use structure, 
 wherein the seal arrangement is configured to be air tight during cell suspension processing, 
 wherein the seal arrangement is configured to be maintained in air contact in the core interior and isolated from contact with any of the cell culture batch during cell suspension processing. 
   
     
     
         2 . The apparatus according to  claim 1  and further comprising:
 a liquid sealing flange in the upper portion,
 wherein the sealing flange projects radially outward in a direction away from the axis, 
 wherein feed cell suspension flow is allowed to enter the core interior below the sealing flange, 
 wherein during cell suspension processing, angular momentum of liquid located in the upper portion above the sealing flange is greater than angular momentum of feed cell suspension entering the core interior below the sealing flange,
 wherein the greater angular momentum generally prevents liquid flow to above the sealing flange. 
 
 
 
     
     
         3 . The apparatus according to  claim 1  and further comprising:
 a rotationally fixed centripetal pump, wherein the centripetal pump is positioned in the upper portion and is in operatively supported connection with the discharge tube, 
 wherein the centripetal pump includes an inlet and an outlet, wherein the inlet is in the core interior vertically below the seal arrangement and in contact with generally cell free centrate during cell suspension processing, and wherein the outlet is in fluid communication with the centrate outlet. 
 
     
     
         4 . The apparatus according to  claim 3   wherein the centrate discharge tube extends annularly outward of and coaxial with the feed tube,
 wherein the centripetal pump is in operative supported connection with the discharge tube. 
   
     
     
         5 . The apparatus according to  claim 4   wherein the centripetal pump comprises a pair of spaced paring discs.   
     
     
         6 . The apparatus according to  claim 5   wherein the feed tube includes an inner feed tube and an outer feed tube,
 wherein the inner feed tube axially extends in the outer feed tube, 
   wherein the paring discs comprise a first paring disc and a second paring disc,
 wherein the first paring disc is attached to the outer feed tube,
 wherein the second paring disc is attached to the centrate discharge tube. 
 
   
     
     
         7 . The apparatus according to  claim 6 ,
 wherein both the first paring disc and the second paring disc extend radially outward away from the axis,
 wherein air isolates from contact with any of the cell culture batch during cell suspension processing, both a lower inner radial portion of the first paring disc wherein the first paring disc is attached to the outer feed tube, and an upper radial portion of the second paring disc where the second paring disc is attached to the centrate discharge tube. 
   
     
     
         8 . The apparatus according to  claim 4  and further comprising
 a rotatable flexible liner, wherein the liner is operatively sealed to the upper flange and extends below the upper flange during cell suspension processing, 
 wherein at least a portion of the liner is radially disposed from the core,
 wherein the core interior is in fluid connection with an area exterior of the core, and wherein the area is fluidly bounded by the liner. 
 
 
     
     
         9 . The apparatus according to  claim 8  wherein the inlet of the centripetal pump is disposed radially outward relative to the axis and away from the centrate discharge tube. 
     
     
         10 . The apparatus according to  claim 8   wherein the seal arrangement includes at least one annular hermetic contact seal.   
     
     
         11 . The apparatus according to  claim 2   and further comprising
 a rotatable flexible liner, wherein the liner is operatively sealed to the upper flange and extends below the upper flange during cell suspension processing, wherein at least a portion of the liner is radially disposed from the core,
 wherein the core interior is in fluid connection with an area exterior of the core, and wherein the area is fluidly bounded by the liner. 
 
   
     
     
         12 . The apparatus according to  claim 11  wherein the upper portion is configured to provide during cell suspension processing, a difference in angular momentum of liquid above and below the sealing flange, wherein a non-contact seal is provided vertically between the sealing flange and the seal arrangement. 
     
     
         13 . The apparatus according to  claim 12  wherein the upper portion includes accelerator fins therein,
 wherein the accelerator fins protrude radially inward in the upper portion, 
 wherein the accelerator fins are disposed above the sealing flange, 
 wherein the accelerator fins are configured to cause during cell suspension processing, the angular momentum of liquid located in the upper portion above the sealing flange, to be greater than angular momentum of feed cell suspension entering below the sealing flange which provides the non-contact seal. 
 
     
     
         14 . The apparatus according to  claim 1  wherein the feed tube comprises an inner feed tube and an outer feed tube, and wherein the inner feed tube axially extends in the outer feed tube. 
     
     
         15 . The apparatus according to  claim 14  wherein the core is configured to receive cell suspension fed through the inner feed tube. 
     
     
         16 . The apparatus according to  claim 6  wherein the feed tube comprises an inner feed tube and an outer feed tube, and wherein the inner feed tube axially extends in the outer feed tube,
 wherein the sealing flange is located at a lower axial end portion of the outer feed tube. 
 
     
     
         17 . The apparatus according to  claim 15  wherein the inner feed tube is axially movable relative to the outer feed tube, and wherein the inner feed tube is usable to remove cell concentrate from a lower area of the single use structure. 
     
     
         18 . The apparatus according to  claim 8  and further comprising:
 a rotatable lower flange, wherein during cell suspension processing the lower flange is axially disposed from the upper flange, 
 wherein the flexible liner is in fluid tight engagement with the upper flange and the lower flange. 
 
     
     
         19 . Apparatus comprising:
 a single use structure for a centrifuge system configured to separate cells in suspension in a cell culture batch, including:
 a rotatable upper flange configured to rotate during cell suspension processing; 
 at least one rotationally fixed feed tube, wherein the feed tube extends along an axis that during cell suspension processing is oriented vertically; 
 a centrate discharge passage extending along the axis, and connected with a centrate outlet; 
 a rotatable core operatively attached to the upper flange and disposed coaxially with respect to the at least one feed tube, wherein the core is configured to receive cell suspension fed through the at least one feed tube, 
 wherein the core includes a core interior, wherein the core has an upper portion and a lower portion and wherein the at least one feed tube extends in the core interior; 
 at least one air tight seal disposed annularly about the at least one feed tube, wherein the at least one seal is in abutting engagement with relatively rotationally movable portions of the single use structure, wherein the at least one seal is configured such that during cell suspension processing the at least one seal is maintained in air contact and isolated from contact with any of the cell culture batch in the interior area. 
   
     
     
         20 . The apparatus according to  claim 19  and further comprising:
 at least one centripetal pump, wherein the at least one centripetal pump is rotationally fixed during cell suspension processing, 
 wherein the at least one centripetal pump includes an inlet and an outlet, 
 wherein the inlet is in fluid connection with liquid in the core interior vertically below air in contact with the at least one seal, 
 and wherein the outlet is in fluid connection with the centrate discharge passage.