Apparatus and method for particle separation in a closed field

• US 5,672,481 A
• Filed: 04/23/1993
• Issued: 09/30/1997
• Est. Priority Date: 10/23/1991
• Status: Expired due to Term

• Current Assignee: Nexell Therapeutics Inc. (Baxter International Inc.)
• Original Assignee: Cellpro, Inc. (Steel Partners, Inc.)
• Inventors: Peterson, Eric, Corpuz, Stanley, Mill, Fred, Thompson, Penny, Maloney, Patrick M., Minshall, Billy W., Heimfeld, Shelly
• Primary Examiner(s): Housel, James C.
• Assistant Examiner(s): FREED, RACHEL
• Application Number:
  US08/052,784
• Time in Patent Office: 1,621 Days
• Field of Search: 435/7.21, 435/2, 435/299, 435/311, 435/961, 435/962, 435/287, 435/316, 436/518, 436/541, 436/176, 436/177, 436/178, 422/44, 422/59, 422/68.1, 210/782, 210/800, 210/103, 604/405, 604/406, 604/409, 604/410, 604/416
• US Class Current: 435/7.21
• CPC Class Codes: A61M 1/0272 Apparatus for treatment of blood or blood constituents prior to or for conservation, e.g. freezing, drying or centrifuging
• CPC Class Codes: A61M 1/0281 Apparatus for treatment of blood or blood constituents prior to transfusion, e.g. washing, filtering or thawing
• CPC Class Codes: A61M 1/3692 Washing or rinsing blood or blood constituents
• CPC Class Codes: B01D 61/147 Microfiltration
• CPC Class Codes: B01D 61/18 Apparatus therefor
• CPC Class Codes: B01D 61/20 Accessories; Auxiliary operations
• CPC Class Codes: B01F 33/45 Magnetic mixers; Mixers with magnetically driven stirrers
• CPC Class Codes: B01F 33/452 using independent floating stirring elements
• CPC Class Codes: B01F 35/20 Measuring; Control or regulation
• CPC Class Codes: B01L 3/50853 with covers or lids
• CPC Class Codes: C12M 47/02 Separating microorganisms from the culture medium; Concentration of biomass separating microorganisms from their culture media C12N1/02
• CPC Class Codes: C12M 47/12 Purification C12M47/04 takes precedence
• CPC Class Codes: G01N 2001/1081 Storing samples under refrigeration
• CPC Class Codes: G01N 2011/0053 using ergometry; measuring power consumption
• CPC Class Codes: G01N 2035/1039 Micropipettes, e.g. microcapillary tubes
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence
• CPC Class Codes: G01N 33/5002 Partitioning blood components
• CPC Class Codes: Y10T 436/2525 Stabilizing or preserving
• CPC Class Codes: Y10T 436/25375 Liberation or purification of sample or separation of material from a sample [e.g., filtering, centrifuging, etc.]
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

A particle separator is provided for collection and manipulation of target particles, e.g., target cells, in a closed sterile field condition. In one embodiment, closed sterile field conditions are maintained from separation through concentration and/or cryo treatment steps and/or transfusion. Preservation of closed sterile field condition are accommodated by using the same integrally coupled rigid-walled vessel for collection and concentration and transfer via integrally coupled conduits to a vessel for cryopreservation and/or transfusion.

In one embodiment, a plurality of valves are responsive to a data processor for controlling the path of fluid flow through the particle separator. A plurality of sensors are provided for providing sensor signals indicative of fluid flowing through the cell separator. A peristaltic pump is responsive to the microprocessor assembly for controlling the speed and direction of fluid flow through the system. A stir plate assembly is responsive to a signal from the data processor for controllably agitating the contents of the cell separator.

Claims

1. A cell separation apparatus having a separation device assembly, the separation device assembly including a separation device for collecting target cells from a sample fluid contained in a sample container, and a fluid control system including a rigid-walled fluid collection vessel for receiving the target cells after being released from the separation device, said cell separation apparatus providing a closed sterile pathway from the sample container to the rigid-walled fluid collection vessel, comprising:

conduits coupling said separation device to said sample container and to a first port of said rigid-walled fluid collection vessel to provide a pathway along which said target cells travel from said sample container to said separation device and from said separation device to said rigid-walled fluid collection vessel in the absence of exposure to the environment, said conduits and said separation device forming a single replaceable item within said separation apparatus;

means responsive to a valve control signal for selectively enabling the fluid coming out of the separation device to flow into the rigid-walled fluid collection vessel; and

means for controlling the operation of the fluid control system, by providing said valve control signal.

2. An apparatus, as claimed in claim 1, wherein said pathway is provided as a closed field sterile pathway.

3. An apparatus, as claimed in claim 1, wherein said separation device is an immunoadsorption column.

4. An apparatus, as claimed in claim 1, wherein said means for controlling comprises a microprocessor.

5. A fluid control system, as claimed in claim 1, further comprising at least a first filter coupled to at least one of said conduits, for filtering the flow through said conduit.

6. A fluid control system, as claimed in claim 5, wherein said first filter is a 0.22 micron filter.

7. A fluid control system, as claimed in claim 5, further comprising a wash solution source, coupled to said separation device wherein said first filter is located between said wash solution source and said separation device.

8. A fluid control system, as claimed in claim 1, further comprising means, between said sample container and to said separation device, for at least partial filteration of fluid which exits said sample container.

9. A fluid control system, as claimed in claim 8, wherein said means for at least partial filteration comprises a means for selectively removing particles ranging from about 5 to about 40 microns.

10. A fluid control system, as claimed in claim 8, wherein said means for at least partly cleaning comprises a pre-column containing a gel.

11. A method for providing separated, concentrated particles from a solution containing said particles, comprising:

providing said solution in a sample container, separated from the environment;

providing a separation device, coupled to said sample container by at least a first conduit, said separation device having an interior separated from the environment;

conveying at least a portion of said solution from said sample container to said interior of said separation device along said first conduit while maintaining said portion of said solution in a closed field condition in the absence of exposure of said portion of said solution to the environment;

retaining at least some of said particles in said interior of said separation device to provide target particles in said separation device;

providing a target vessel, coupled to said interior of said separation device by at least a second conduit, said target vessel having substantially rigid walls and a plurality of ports, each said port being sufficiently sealable from the environment to define a closed-field vessel, wherein at least a first of said plurality of ports of said target vessel is sealed by a filter;

conveying a plurality of said target particles from said interior of said separation device to said target vessel while maintaining said plurality of target particles in a closed sterile field condition in the absence of exposure of said plurality of target particles to the environment; and

concentrating said target particles without removing said target particles from said target vessel, to maintain said plurality of target particles in a closed sterile field condition.

12. A method, as claimed in claim 11, wherein said step of concentrating comprises centrifuging said plurality of target particles.

13. A method, as claimed in claim 11, further comprising expelling at least some of the contents of said collection vessel during said step of conveying a plurality of said target particles.

14. A method, as claimed in claim 13, wherein said step of expelling comprises venting gaseous contents of said collection vessel through a filter.

15. A method, as claimed in claim 14, wherein said filter is about a 0.22 micron filter.

16. A method, as claimed in claim 13, further comprising:

cryo-treating said plurality of target particles without removing said target particles from said collection vessel, to maintain said plurality of target particles in a closed field condition.

17. The method of claim 11 wherein said separation device further comprises:

an interior of said device containing a substrate having an affinity for a target particle;

said interior of said device containing a protein substantially coating at least said substrate and interior surface of said device, prior to coupling said separation device to a particle separation apparatus.

18. The separation device of claim 17, wherein said protein comprises human serum albumen.

19. The separation device of claim 17, wherein said protein is irradiated protein.

20. A method for contamination-free, repeatable particle separation comprising:

a) providing a particle separation device having a separation column receptacle and a plurality of controllable valves;

b) providing a separation device having an affinity-specific substrate, and said separation device having an inlet port and an outlet port;

c) mounting at least first and second conduits, coupled to said inlet and outlet ports, respectively in operable proximity to said controllable valves to permit selective opening and closing of said conduits;

d) coupling said conduits to at least a sample source and a target collection vessel, said collection vessel having substantially rigid walls and a plurality of ports, each said port being sufficiently sealable from the environment to define a closed-field vessel, wherein at least a first of said plurality of ports of said collection vessel is sealed by a filter;

e) controlling said controllable valves to convey contents of said sample source to said separation device wherein at least a first plurality of target particles are retained in said separation device;

f) controlling said controllable valves to convey said first plurality of target particles to said collection vessel;

g) removing said sample source, said conduits, said separation device and said collection vessel;

h) repeating steps b) through g), using a different sample source, conduits, separation device and collection vessel to separate a second plurality of target particles in the absence of contamination by said first plurality of target particles or by the contents of a previous sample source.

21. A method, as claimed in claim 20, wherein said step of providing a separation device includes coating the interior and contents of said separation device with a protein, prior to said step of mounting said separation device.

22. A method, as claimed in claim 21, wherein said step of coating further includes irradiating said protein.

23. A method, as claimed in claim 22, wherein said irradiation comprises irradiating with an electron beam.

24. A method, as claimed in claim 20, wherein said steps of mounting and repeating are performed substantially in the absence of splicing said conduits.