Rigid chamber for cell separation from a flexible dissposable bag

Abstract

Method consists of placing a flexible container within a rigid frame and expanding the container by pneumatic or hydraulic pressure such that the walls of the container conform to the inside walls of the rigid frame thus forming a well-defined chamber. The system has the capability of reducing the volume of the chamber by adjusting the distance between the walls of the rigid container. The methods and systems so described are applicable to closed sterile systems that employ immunomagnetic isolation or purging of components from blood products. By providing a fixed volume and at least one surface upon which targeted entities can be magnetically deposited, target cells in the case of positive isolations can be magnetically held, flushed with wash buffers over them to remove entrapped cells and finally the recovery of product of extremely high purities and at high yields.

We claim: 
     
         1 . A cell separation system, comprising:
 a rigid frame having oppositely facing wall members defining a confinement gap;   a flexible container disposed in said confinement gap, said flexible container having a first end and a second end and a pair of expansible panels joined along a periphery thereof such that facing interior surfaces of said panels form a collection space with an opening at each of said ends, and said flexible panels being expansible under the influence of positive pressure applied within said collection space to urge said panels into engagement with said oppositely facing wall members within said confinement gap and thereby render said container substantially inflexible;   a valved inlet port at said first end and a valved outlet port at said second end that allow controlled fluid flow through said container and that allow the positive pressure within the container to be maintained during fluid flow; and   a pressure source operatively connected to said container for applying positive pressure within said collection space;   wherein at least one of said wall members includes a magnet component effective to generate a magnetic field gradient in said collection space; and   wherein the system is arranged to magnetically separate labeled target cells from a mixed cell population within the container by and under the influence of the magnetic field gradient generated by the magnet component and to apply said positive pressure to expand said flexible container and maintain said positive pressure throughout magnetic cell separation of the labeled target cells from the mixed cell population so as to maintain said container inflexible throughout a cell separation process.   
     
     
         2 . The system according to  claim 1 , further comprising a pressure relief valve operatively connected to said container and effective to maintain the pressure within said container at a predetermined pressure. 
     
     
         3 . The system according to  claim 1 , further comprising a pressure gauge configured to monitor pressure inside said container. 
     
     
         4 . The system according to  claim 1 , wherein one of said oppositely facing wall members of said rigid frame is movable in relation to the other, whereby said confinement gap is adjustable. 
     
     
         5 . The system according to  claim 1 , wherein both of said oppositely facing wall members of said rigid frame are movable in relation to each other, whereby said confinement gap is adjustable. 
     
     
         6 . The system according to  claim 1 , wherein said magnet component is reversibly disengageable from said at least one oppositely facing wall member such that said magnetic field gradient in said collection space is able to be reversibly attenuated. 
     
     
         7 . The system according to  claim 1 , wherein each of said oppositely facing wall members of said rigid frame comprises a magnet component effective to generate a magnetic field gradient in said collection space. 
     
     
         8 . The system according to  claim 7 , wherein said magnet component is reversibly disengageable from each of said oppositely facing wall members such that said magnetic field gradient in said collection space is able to be reversibly attenuated. 
     
     
         9 . A method of separating magnetically labeled target cells from a mixed cell population with the cell separating system according to  claim 1 , comprising the steps of:
 placing the flexible container into the confinement gap of the rigid frame;   expanding the flexible container by application of positive pressure such that exterior surfaces of said panels are urged into engagement with the oppositely facing wall members within the confinement gap, thereby rendering the container substantially inflexible; and   separating magnetically labeled target cells from a mixed cell population within the container by and under the influence of the magnetic field gradient generated by the magnet component, the positive pressure applied to expand the flexible container being maintained throughout the separating step so as to maintain the container inflexible throughout the separating step.   
     
     
         10 . The method according to  claim 9 , wherein the flexible container is a sterile, disposable bag adapted for aseptically processing biological entities. 
     
     
         11 . The method according to  claim 9 , wherein the container is expanded by applying hydraulic pressure. 
     
     
         12 . The method according to  claim 9 , wherein the oppositely facing wall members of said rigid frame are generally planar and substantially parallel to one another. 
     
     
         13 . The method according to  claim 9 , further comprising the step of adjusting a distance from which the oppositely facing wall members are spaced within the confinement gap. 
     
     
         14 . The method according to  claim 9 , further comprising the steps of:
 before said separating step, introducing the mixed cell population and at least one magnetic labeling agent into the collection space of the container, the at least one magnetic labeling agent being effective to selectively bind to and magnetically label the target cells; and   after said separating step, recovering the magnetically labeled target cells.   
     
     
         15 . The method according to  claim 14 , wherein, during said separating step, a wash fluid is passed through the container at least once before said recovering step. 
     
     
         16 . The method according to  claim 14 , wherein the mixed cell population and the at least one magnetic labeling agent are simultaneously introduced into the collection space. 
     
     
         17 . The method according to  claim 14 , wherein the mixed cell population and the at least one magnetic labeling agent are sequentially introduced into said collection space. 
     
     
         18 . The method according to  claim 9 , further comprising the step of disengaging and reversing the magnet component relative to the at least one of said wall members such that the magnetic field gradient in the collection space is reversibly attenuated. 
     
     
         19 . The method according to  claim 9 , wherein each of the oppositely facing wall members of said rigid frame comprises a magnet component effective to generate the magnetic field gradient in the collection space, and further comprising the step of disengaging and reversing the magnet component on each of the oppositely facing wall members such that the magnetic field gradient in the collection space is reversibly attenuated.