Method using an apparatus for separation of biological fluids

• US 6,291,249 B1
• Filed: 02/28/2000
• Issued: 09/18/2001
• Est. Priority Date: 03/02/1999
• Status: Expired due to Term

• Current Assignee: Qualigen, Inc.
• Original Assignee: Qualigen, Inc.
• Inventors: Mahant, Vijay, Doneen, Byron
• Primary Examiner(s): Le, Long V.
• Assistant Examiner(s): GABEL, GAILENE
• Application Number:
  US09/514,686
• Time in Patent Office: 568 Days
• Field of Search: 422/56-61, 422/44, 422/48, 422/77, 422/101, 422/102, 435/7.2, 435/7.23, 435/7.24, 435/7.25, 435/287.9, 435/288--28, 436/177, 436/178, 436/520, 436/526-531, 436/535, 436/547, 436/810, 436/825, 436/826, 210/222, 210/223, 210/451
• US Class Current: 436/177
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 3/5021 Test tubes specially adapted for centrifugation purposes centrifuges B04B5/04
• CPC Class Codes: B01L 3/505 flexible containers not provided for above
• CPC Class Codes: B03C 1/01 by addition of magnetic adjuvants
• CPC Class Codes: B03C 1/288 disposed at the outer circumference of a recipient
• CPC Class Codes: B03C 1/30 Combinations with other devices, not otherwise provided for
• 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 applications
• CPC Class Codes: G01N 2001/4016 being a selective membrane, e.g. dialysis or osmosis
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence
• CPC Class Codes: Y10S 436/807 Apparatus included in process claim, e.g. physical support structures
• CPC Class Codes: Y10T 436/113332 with conveyance of sample along a test line in a container or rack
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials
• 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
• CPC Class Codes: Y10T 436/2575 Volumetric liquid transfer

Abstract

A cell containing sample is separated into a cell containing portion and a substantially cell depleted portion, by mixing the sample with particles to produce a cell containing network, and separating the network from the remaining substantially cell depleted portion within a plurality of confining walls, wherein at least one of the walls is flexible. While in some aspects the separation is performed employing a magnetic force, in other aspects the separation is performed using two forces, wherein one force is a magnetic force and the other force is a mechanical force. It is contemplated that whole blood may be used as the sample, and that the cell-containing portion largely comprises a network of inter-linked red blood cells. It is especially contemplated that separation involves antiligands, preferably primary antibodies that bind to a ligand such as antigen on or in red blood cell membranes, and secondary antibodies that bind to the primary antibodies. It is also contemplated that the primary antibodies can be coupled to the surfaces of paramagnetic beads.

Claims

1. A method of separating a sample containing blood cells into a cell-containing portion and a cell-depleted portion comprising:

receiving the sample in a vessel, wherein the vessel retains the sample within a plurality of confining walls, and wherein at least one of the confining walls is flexible

providing an additive and magnetic particles, wherein the additive binds to the cells and the magnetic particles and wherein the magnetic particles bind to the cells and the additive;

combining the sample, the additive, and the magnetic particles to produce a cell-containing network; and

separating the network within the plurality of confining walls from the cell-depleted portion by applying a first and a second force, wherein the first force is a magnetic force and the second force is an automatic mechanical force that is transmitted through at least one of the confining walls.

2. The method of claim 1 wherein the sample received in the vessel has a volume of no less than about 3 ml.

3. The method of claim 1 wherein the sample received in the vessel has a volume of less than about 1 ml.

4. The method of claim 1 wherein the mechanical force comprises a pressure that is applied to the at least one flexible confining wall.

5. The method of claim 1 wherein each of the magnetic particles has a mean volume of between about 5×

10<sup>−

24</sup> m³ to about 5×

10<sup>−

6</sup> m³.

6. The method of claim 1 wherein the particles are coated with a coating.

7. The method of claim 6 wherein the coating comprises an anti-ligand.

8. At The method of claim 6 wherein the coating comprises an antibody.

9. The method of claim 6 wherein the coating comprises a polycationic polymer.

10. The method of claim 1 wherein the binding of the additive results from the additive comprising an anti-ligand.

11. The method of claim 1 wherein the magnetic particles comprise a primary antibody and the additive comprises a secondary antibody, and wherein the primary antibody binds to a surface component of the cells, and the secondary antibody binds to the primary antibody.

12. The method of claim 1 wherein the cells predominantly comprise red blood cells.

13. The method of claim 1 wherein the blood cells comprise white blood cells and platelets.

14. The method of claim 1, further comprising measuring prostate specific antigen.

15. The method of claim 1, further comprising measuring creatinine.

16. The method of claim 1 wherein at least 70% by volume of the cell depleted portion is separated from the network within ten minutes.

17. The method of claim 1 wherein separating the network produces a separation efficiency of at least 70%.

18. The method of claim 1 wherein separating the network produces a separation efficiency of at least 80%.

19. The method of claim 1 wherein separating the network produces a separation efficiency of at least 90%.

20. The method of claim 1 wherein at least 90% by volume of the cell depleted portion is separated from the network within ten minutes, with a separation efficiency of at least 95%.