Sample processing system and methods

• US 8,727,132 B2
• Filed: 12/08/2008
• Issued: 05/20/2014
• Est. Priority Date: 12/07/2007
• Status: Active Grant

• Current Assignee: Miltenyi Biotec B.V. & Co. KG (Miltenyi GmbH)
• Original Assignee: Miltenyi Biotec GmbH
• Inventors: Miltenyi, Stefan, Schimmelpfennig, Winfried, Lantow, Holger, Biehl, Martin, Kabaha, Eiad, Schulz, Jrgen, Neuschfer, Elmar Niklas
• Primary Examiner(s): BOWERS, NATHAN ANDREW
• Application Number:
  US12/745,688
• Publication Number:
  US 20110003380A1
• Time in Patent Office: 1,989 Days
• US Class Current: 210/360.1
• CPC Class Codes: A61K 39/4611 T-cells, e.g. tumor infiltrating lymphocytes [TIL], lymphokine-activated killer cells [LAK] or regulatory T cells [Treg]
• CPC Class Codes: A61K 39/4615 Dendritic cells
• CPC Class Codes: A61K 39/4621 immunosuppressive or immunotolerising
• CPC Class Codes: A61K 39/4622 Antigen presenting cells
• CPC Class Codes: A61K 39/46433 Antigens related to auto-immune diseases; Preparations to induce self-tolerance
• CPC Class Codes: A61K 39/46434 Antigens related to induction of tolerance to non-self
• CPC Class Codes: A61K 39/4644 Cancer antigens
• CPC Class Codes: A61K 39/464838 Viral antigens
• CPC Class Codes: A61M 1/0209 Multiple bag systems for separating or storing blood components
• CPC Class Codes: A61M 1/3618 Magnetic separation
• CPC Class Codes: A61M 1/3623 Means for actively controlling temperature of blood
• CPC Class Codes: A61M 1/3633 Blood component filters, e.g. leukocyte filters
• CPC Class Codes: A61M 1/3679 by absorption A61M1/3675 takes precedence
• CPC Class Codes: A61M 1/3692 Washing or rinsing blood or blood constituents
• CPC Class Codes: A61M 1/3693 using separation based on different densities of components, e.g. centrifuging
• CPC Class Codes: A61M 1/3696 with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: A61M 2202/0071 product to be retained or harvested, e.g. by pheresis
• CPC Class Codes: A61M 2202/0413 Blood
• CPC Class Codes: A61M 2202/0427 Platelets; Thrombocytes
• CPC Class Codes: A61M 2202/10 Bone-marrow
• CPC Class Codes: B01D 15/3809 : of the antigen-antibody type, e.g. protein A, G, L chromatography
• CPC Class Codes: B01D 15/3885 : Using electrical or magnetic means
• CPC Class Codes: B01D 21/26 : Separation of sediment aided by centrifugal force or centripetal forcecentrifuges B04B; cyclones B04C
• CPC Class Codes: B01D 21/262 : by using a centrifuge
• CPC Class Codes: B01L 3/5021 : Test tubes specially adapted for centrifugation purposes centrifuges B04B5/04
• CPC Class Codes: B03C 1/002 : High gradient magnetic separation acting directly on the substance being separated B03C1/025; acting on the medium containing the substance being separated B03C1/32
• CPC Class Codes: B03C 1/027 : with reciprocating canisters
• CPC Class Codes: B03C 1/032 : Matrix cleaning systems
• CPC Class Codes: B03C 1/0332 : using permanent magnets
• CPC Class Codes: B03C 1/0335 : using coils
• CPC Class Codes: B03C 1/034 : characterised by the matrix elements
• CPC Class Codes: B03C 1/288 : disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 2201/18 : Magnetic separation whereby the particles are suspended in a liquid
• CPC Class Codes: B03C 2201/26 : for use in medical or biological applications
• CPC Class Codes: B04B 1/12 : with continuous discharge
• CPC Class Codes: B04B 13/00 : Control arrangements specially designed for centrifuges; Programme control of centrifuges control arrangements for feed, charge, or discharge B04B11/00
• CPC Class Codes: B04B 2005/0464 : with hollow or massive core in centrifuge bowl
• CPC Class Codes: B04B 2013/006 : Interface detection or monitoring of separated components
• CPC Class Codes: B04B 5/00 : Other centrifuges
• CPC Class Codes: B04B 5/04 : Radial chamber apparatus for separating predominantly liquid mixtures, e.g. butyrometers
• CPC Class Codes: B04B 5/0442 : with means for adding or withdrawing liquid substances during the centrifugation, e.g. continuous centrifugation
• CPC Class Codes: B04B 5/10 : Centrifuges combined with other apparatus, e.g. electrostatic separators; Sets or systems of several centrifuges B04B5/12 takes precedence; magnetic or electrostatic separators B03C; amassing particles by electric fields, e.g. by agglomeration B03C3/0175
• CPC Class Codes: B04B 7/08 : Rotary bowls centrifugal casting machines B22D
• CPC Class Codes: C12M 23/28 : disposable or single use
• CPC Class Codes: C12M 33/10 : by centrifugation centrifuges in general B04B; Cyclones cyclones in general B04C
• CPC Class Codes: C12M 41/48 : Automatic or computerized control automatic analysis G01N35/00
• CPC Class Codes: C12M 47/04 : Cell isolation or sorting purging biological preparations of unwanted cells C12N5/0081, determining the presence or kind of microorganism C12Q1/04
• CPC Class Codes: C12N 2500/34 : Sugars
• CPC Class Codes: C12N 2501/998 : Proteins not provided for elsewhere
• CPC Class Codes: C12N 5/0018 : Culture media for cell or tissue culture media for specific animal cell type C12N5/06
• CPC Class Codes: C12N 5/0636 : T lymphocytes
• CPC Class Codes: C12N 5/0646 : Natural killers cells [NK], NKT cells
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: C12N 5/0669 : Bone marrow stromal cells; Whole bone marrow isolated stem cells from bone marrow C12N5/0647, C12N5/0663
• CPC Class Codes: G01N 2333/70596 : Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
• CPC Class Codes: G01N 33/58 : involving labelled substances G01N33/53 takes precedence
• CPC Class Codes: G01N 35/0098 : involving analyte bound to insoluble magnetic carrier, e.g. using magnetic separation magnetic particles used in immunoassays G01N33/54326; magnetic separation in general B03C

Abstract

The invention relates to a system, comprising: a) a sample processing unit, comprising an input port and an output port coupled to a rotating container having at least one sample chamber, the sample processing unit configured provide a first processing step to a sample or to rotate the container so as to apply a centrifugal force to a sample deposited in the chamber and separate at least a first component and a second component of the deposited sample; and b) a sample separation unit coupled to the output port of the sample processing unit, the cell separation unit comprising separation column holder (42), a pump (64) and a plurality of valves (1-11) configured to at least partially control fluid flow through a fluid circuitry and a separation column (40) positioned in the holder, the separation column configured to separate labeled and unlabeled components of sample flowed through the column.

Claims

1. A system for separating cells, comprising:

a) a sample processing unit, comprising an input port, an output port, and a centrifuge chamber,

wherein the centrifuge chamber comprises a circular bottom, a circular top, and a wall passing therebetween, thus forming an internal compartment that is configured to contain and process a biological sample comprising cells;

wherein the sample processing unit is configured to rotate the centrifuge chamber about an axis that passes through the circular bottom and the circular top so as to apply a centrifugal force to a sample deposited in the chamber and thereby to separate at least a first component and a second component of the deposited sample;

wherein the input port and the output port are configured and positioned to deliver a sample to and from the internal compartment while the centrifuge chamber is rotating;

wherein the centrifuge chamber comprises a detection window that extends radially in the bottom or top of the chamber from a position adjacent to the rotation axis to a position near the perimeter of the chamber; and

wherein the sample processing unit further comprises a detector that is constructed and aligned to receive light passing through a sample in the detection window of the centrifuge chamber so as to assess a characteristic of the deposited sample radially through and along the detection window, thereby monitoring separation of the first component from the second component in the chamber;

the system further comprising;

b) a sample separation unit coupled to the output port of the sample processing unit, the sample separation unit comprising;

a separation column holder,

a pump,

a plurality of valves configured to at least partially control fluid flow through a fluid circuitry, and

a separation column positioned in the holder, wherein the separation column is configured to separate labeled and unlabeled components of a sample flowed through the column.

2. The system according to claim 1, configured so that light from a light source can at least partially penetrate through at least a part of a sample that is being separated in the centrifuge chamber through the detection window, and light passing through at least part of the sample can be detected by a light detector.

3. The system according to claim 1, wherein the detector is positioned such that it can monitor separation of the components in the centrifuge chamber through the detection window from an area adjacent to the rotation axis of the centrifuge chamber to an area adjacent to the perimeter of the container.

4. The system according to claim 1, wherein a prism or a mirror is positioned radially along the detection window.

5. The system according to claim 4, wherein the prism or mirror is positioned to cover a channel that is positioned in or near a lid of the centrifuge chamber, wherein the channel is configured such that at least a part of the sample can flow into it during centrifugation.

6. The system according to claim 1, wherein the centrifuge chamber comprises a plurality of layered surfaces between the top and the bottom and perpendicular to the rotational axis upon which cells may be cultured.

7. The system according to claim 6, wherein the centrifuge chamber comprises at least one layer of growing cells thereon.

8. The system according to claim 7 wherein the centrifuge chamber is disposable and/or can be sterilized.

9. The system according to claim 1, wherein the centrifuge chamber comprises a material chosen from the group consisting of:

plastics, polystyrol (PS), polystyrene, polyvinylchloride, polycarbonate, glass, polyacrylate, polyacrylamide, polymethylmethacrylate (PMMA), polyethylene terephthalate (PET), polytetrafluoroethylene (PTFE), thermoplastic polyurethane (TPU), silicone, polyethylene (PE), collagen, chitin, alginate, hyaluronic acid derivatives, polylactide (PLA), polyglycolide (PGA) and their copolymers, polystyrol (PS), polycarbonate, polyacrylate, ceramics, hydroxyapatite (HA), calcium phosphate, and combinations thereof.

10. The system according to claim 1, wherein the inlet port of the sample processing unit is connected to the internal compartment of the centrifuge chamber by way of a central line that passes through rotational seals and along the rotational axis.

11. The system according to claim 1, wherein both the inlet port and the outlet port of the sample processing unit are connected to the internal compartment of the centrifuge chamber by way of an inner line and an outer line that both pass through rotational seals and along the rotational axis.