Sequential processing of biological fluids

Abstract

A process for the sequential processing of opaque and transparent biological fluids such as whole blood, apheresis blood, bone marrow blood, umbilical cord blood, buffy coat or cultured cells by processing steps in a hollow cylindrical centrifugal processing chamber ( 300 ) which is part of a disposable set. At least three different procedures selected from washing, incubation, transduction, separation, density gradient separation, dilution and volume adjustment are each carried out once or repeated a number of times according to a given processing profile in the processing chamber. Each procedure involves an input into the processing chamber, an operation in the processing chamber and an output from the processing chamber by displacement of a piston ( 310 ). The at least three different procedures are sequentially chained one after the other to constitute an overall sequential operation in the processing chamber and its disposable set. A first application is incubation for binding magnetic beads with human blood cells or stem cells. A second application is transduction by which foreign genetic material is inserted into human blood cells or stem cells by a virus. A third application is reconditioning biological fluids to achieve reproducible concentration and volumes of blood cells or stem cells.

1 . A process for the sequential processing of opaque and transparent biological fluids such as whole blood, apheresis blood, bone marrow blood, umbilical cord blood, buffy coat or cultured cells by processing steps selected from washing, incubation, transduction, separation, density gradient separation, dilution and volume adjustment in a hollow cylindrical centrifugal processing chamber for processing or mixing the biological fluids and/or mixing of the biological fluids with reagents and/or diluents, wherein the centrifugal processing chamber contains within its generally cylindrical wall an axially movable member such as a piston which is axially movable to vary the volume of a separation space within the processing chamber to intake or to output biological fluids into or from the separation space, wherein the processing chamber is part of a disposable set containing the biological fluids and reagents and/or diluents, the disposable set constituting an enclosed sterile environment, characterized in that, using a single processing chamber as part of a single disposable set, at least three different procedures selected from washing (including pre-washing and post-washing as separate procedures), incubation, transduction, separation, density gradient separation, dilution and volume adjustment are carried out; each said procedure being carried out once or repeated a number of times according to a given processing profile in the processing chamber of the single disposable set; each said procedure involving an input into the processing chamber produced by axially moving the movable member, an operation in the processing chamber and an output from the processing chamber produced by axially moving the movable member; the at least three different procedures being sequentially chained one after the other in the processing chamber to constitute a chained sequence of the different procedures as an overall sequential operation in the processing chamber and its single disposable set, all of the biological fluids, reagents and/or diluents necessary for carrying out all of said at least three different procedures being contained in the enclosed sterile environment of the single disposable set used for the overall sequential operation. 
     
     
         2 . A process as claimed in  claim 1 , wherein the chained sequence constituting an overall sequential operation in the processing chamber comprises one of the following sequences of procedures (i), (ii) and (iii):
 (i) pre-washing; addition of reagents and incubation; post-washing,   (ii) washing; dilution; centrifugal separation; dilution; volume adjustment of the product, and   (iii) preliminary thawing; washing; sampling and dilution; volume adjustment/extraction.   
     
     
         3 . A process as claimed in  claim 2 , wherein the following procedures are carried out sequentially: pre-washing; addition of reagents and incubation; post-washing. 
     
     
         4 . A process as claimed in  claim 3 , which is for binding magnetic beads with human blood cells or stem cells. 
     
     
         5 . A process as claimed in  claim 2 , wherein the following procedures are carried out sequentially: washing; dilution; centrifugal separation; dilution; volume adjustment of the product. 
     
     
         6 . A process as claimed in  claim 5 , which is for virus transduction with lymphocytes T-cells or other human cells. 
     
     
         7 . A process as claimed in  claim 2 , wherein the following procedures are carried out sequentially: preliminary thawing; washing; sampling and dilution; volume adjustment/extraction. 
     
     
         8 . A process as claimed in  claim 7 , which is for recondition dosing of multiple doses of cellular products. 
     
     
         9 . A process as claimed in  claim 1 , wherein the position of the movable member is monitored by an infrared sensor. 
     
     
         10 . A process as claimed in  claim 1 , wherein the volume of liquids treated in the processing chamber of one disposable set for one overall sequential operation is from four to sixteen times the maximum treatment volume of the processing chamber. 
     
     
         11 . A process as claimed in  claim 1 , wherein the processing chamber is associated with two external pinch valves controlling tubing of the disposable set for selecting two biological additives for switching bags of the disposable set without user intervention.