Devices and methods for separating cellular components of blood from liquid portion of blood

• US 6,069,014 A
• Filed: 08/19/1997
• Issued: 05/30/2000
• Est. Priority Date: 05/09/1995
• Status: Expired due to Fees

• Current Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Original Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Inventors: Jaseph, Corey L., Schrier, Wayne H., Ruggeri, Franco, Schoengold, Ronald J.
• Primary Examiner(s): Chin, Christopher L.
• Application Number:
  US08/914,426
• Time in Patent Office: 1,015 Days
• Field of Search: 422/56, 422/58, 422/101, 435/2, 435/4, 435/7.1, 435/7.92, 435/287.1, 435/287.2, 435/287.7, 435/287.8, 435/287.9, 435/805, 435/810, 435/962, 435/97, 436/514, 436/518, 436/169, 436/170, 436/177, 436/178, 436/808, 436/810
• US Class Current: 436/518
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: G01N 2333/205 from Campylobacter (G)
• CPC Class Codes: G01N 30/02 Column chromatography
• CPC Class Codes: G01N 30/90 Plate chromatography, e.g. thin layer or paper chromatography
• 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: G01N 33/521 Single-layer analytical elements
• CPC Class Codes: G01N 33/525 Multi-layer analytical elements
• CPC Class Codes: G01N 33/54386 Analytical elements
• CPC Class Codes: G01N 33/54388 based on lateral flow
• CPC Class Codes: G01N 33/54389 with bidirectional or multidirectional lateral flow, e.g. wherein the sample flows from a single, common sample application point into multiple strips, lanes or zones
• CPC Class Codes: Y10S 435/805 Test papers
• CPC Class Codes: Y10S 435/81 Packaged device or kit
• CPC Class Codes: Y10S 435/962 Prevention or removal of interfering materials or reactants or other treatment to enhance results, e.g. determining or preventing nonspecific binding
• CPC Class Codes: Y10S 435/97 Test strip or test slide
• CPC Class Codes: Y10S 436/808 Automated or kit
• CPC Class Codes: Y10S 436/81 Tube, bottle, or dipstick
• 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 device for the separation of the liquid portion of blood from the cellular components of blood comprises: (1) a pad of porous material permeable to the liquid portion of blood but capable of trapping the cellular components of blood; (2) a substrate supporting the pad; and (3) means, attached to the pad, for facilitating the flow of the liquid portion of the blood: (i) through interstices around the trapped cellular components of the blood and (ii) from the pad of porous material. The separation of the liquid portion of blood from the cellular components of the blood occurs by flow through the pad of porous material without significant hemolysis. The device can be incorporated into a device for the performance of specific binding assays such as immunoassays. The pad of porous material can contain an agglutinating agent such as a lectin or an anti-blood cell antibody, or a carbohydrate such as mannitol. Other devices and methods for the separation of the liquid portion of blood from the cellular components of the blood are also disclosed.

Claims

1. A device for separating the liquid portion of blood from the cellular components of blood comprising:

(a) a first opposable component including;

(i) a first porous separating matrix permeable to the liquid portion of blood but capable of trapping the cellular components of blood; and

(ii) a second porous matrix in operable contact with the first porous separating matrix that permits the liquid portion of the blood to flow by capillary action through the second porous matrix; and

(b) a second opposable component attachable to the first opposable component for applying fluid to the first opposable component such that the first and second opposable components can be brought into opposition to transfer fluid from the second opposable component to the first opposable component by pressure;

whereby the separation of the liquid portion of blood from the cellular components of the blood occurs by flow through the first and second matrices of the first opposable component without significant hemolysis.

2. The device of claim 1 wherein the first porous separating matrix contains a binder for the cellular components of blood.

3. The device of claim 2 wherein the binder is an anti-blood cell antibody.

4. The device of claim 3 wherein the anti-blood cell antibody is an anti-erythrocyte antibody.

5. The device of claim 2 wherein the binder is a lectin.

6. The device of claim 5 wherein the lectin is selected from the group consisting of concanavalin A, abrin, phytohaemagglutinin, limulin, and the lectins produced by the following species:

Agaricus bisporus, Anguilla anguilla, Arachis hypogaea, Bandeiraea simplicifolia, Bauhinia purpurea, Caragana arborescens, Cicer arietinum, Codium fragile, Datura stramonium, Dolichos biflorus, Erythrina corallodendron, Erythrina cristagalli, Euonymus europaeus, Glycine max, Helix aspersa, Helix pomatia, Lathyrus odoratus, Lens culinaris, Lycopersicon esculentum, Maclura pomifera, Momordica charantia, Mycoplasma gallisepticum, Naja mocambique, Naja kaouthia, Perseau americana, Phaseolus coccineus, Phaseolus limensis, Phaseolus vulgaris, Phytolacca americana, Pisum sativum, Pseudomonas aeruginosa, Psophocarpus tetragonolobus, Ptilota plumosa, Ricinus communis, Robinia pseudoacacia, Sambucus nigra, Solanum tuberosum, Sophora japonica, Tetragonolobus purpureas, Triticum vulgaris, Ulex europaeus, Vicia faba, Vicia sativa, Vicia villosa, Vigna radiata, Viscum album, and Wisteria floribunda.

7. The device of claim 1 wherein the first porous separating matrix is impregnated with a carbohydrate capable of aggregating blood cells.

8. The device of claim 7 wherein the carbohydrate is selected from the group consisting of mannitol, sorbitol, inositol, β

-D-glucose, α

-D-glucose, D(+)xylose, D(+)mannose, D(-)arabinose, L(+)arabinose, D(+)galactose, L(-)xylose, D-glucoheptose, L-lyxose, lactose, maltose, and sucrose.

9. The device of claim 8 wherein the carbohydrate is mannitol.

10. The device of claim 1 wherein the second matrix is a membrane for chromatographic separation.

11. The device of claim 10 wherein the membrane for chromatographic separation has a capture zone thereon for binding a member of a specific binding pair.

12. The device of claim 1 wherein the first porous separating matrix is an asymmetric membrane with a first surface and a second surface, the membrane having a gradient of pore sizes such that the pore size decreases from the first surface to the second surface, the asymmetric membrane capable of trapping the cellular components of blood within it and allowing the liquid components of blood to pass through.

13. The device of claim 1 wherein the first porous separating matrix includes two sectors:

(i) a first sector permeable to both the liquid portion of blood and the cellular components of blood; and

(ii) a second sector permeable to the liquid portion of blood but capable of binding the cellular components of blood.

14. The device of claim 1 wherein the second opposable component includes a sample application zone for transferring fluid to the first opposable component, the fluid including a blood sample.

15. The device of claim 14 wherein the sample application zone for transferring fluid from the second opposable component to the first opposable component includes at least one reagent for treatment of the sample.

16. The device of claim 14 wherein the sample application zone on the second opposable component includes a specific binding partner labeled with a detectable label, the specific binding partner having specific binding affinity for at least one component selected from the group consisting of the analyte and a specific binding partner for the analyte in a form that can be resolubilized by the addition of an aqueous sample to the sample application zone on the second opposable component.

17. The device of claim 16 wherein the specific binding partner labeled with the detectable label has specific binding affinity for the analyte.

18. A method for performing an assay for detecting or determining at least one analyte in liquid portion of a blood sample comprising the steps of:

(a) applying a sample of blood to the sample application zone on the second opposable component of the device of claim 11 to resolubilize a labeled specific binding partner in the sample application zone;

(b) bringing the first and second opposable components into opposition to transfer fluid from the second opposable component to the first opposable component by pressure, the fluid including the blood sample and the resolubilized labeled specific binding partner;

(c) allowing the blood sample to flow through the first porous separating matrix to separate the liquid portion of the blood sample from the cellular components of the blood sample;

(d) facilitating the flow of the liquid portion of blood through the interstices around the trapped cellular components of the blood as a result of the action of the second matrix; and

(e) allowing the liquid portion of the blood to flow through the second matrix so that an assay is performed in the second matrix, the assay being performed by binding a member of a specific binding pair that captures onto the second matrix to detect or determine the at least one analyte, at least one member of the specific binding pair binding to the analyte.

19. A device for separating the liquid portion of blood from the cellular components of blood comprising:

(a) a first opposable component including a porous pad for application of a blood sample, wherein the porous pad includes a first porous separating matrix permeable to the liquid portion of blood but capable of trapping the cellular components of blood; and

(b) a second opposable component attachable to the first opposable component so that the first and second opposable components can be brought into opposition to transfer fluid from the first opposable component to the second opposable component by pressure, the second opposable component for receiving fluid from the first opposable component, the second opposable component including therein a second porous matrix in operable contact with the first porous separating matrix when the first and second opposable components are brought into opposition, the second porous matrix permitting the liquid portion of the blood to flow by capillary action through the second porous matrix;

whereby the separation of the liquid portion of the blood from the cellular components of the blood occurs by flow through the first and second matrices without significant hemolysis.

20. The device of claim 19 wherein the first porous separating matrix contains a binder for the cellular components of blood.

21. The device of claim 20 wherein the binder is an anti-blood cell antibody.

22. The device of claim 21 wherein the anti-blood cell antibody is an anti-erythrocyte antibody.

23. The device of claim 20 wherein the binder is a lectin.

24. The device of claim 23 wherein the lectin is selected from the group consisting of concanavalin A, abrin, phytohaemagglutinin, limulin, and the lectins produced by the following species:

Agaricus bisporus, Anguilla anguilla, Arachis hypogaea, Bandeiraea simplicifolia, Bauhinia purpurea, Caragana arborescens, Cicer arietinum, Codium fragile, Datura stramonium, Dolichos biflorus, Erythrina corallodendron, Erythrina cristagalli, Euonymus europaeus, Glycine max, Helix aspersa, Helix pomatia, Lathyrus odoratus, Lens culinaris, Lycopersicon esculentum, Maclura pomifera, Momordica charantia, Mycoplasma gallisepticum, Naja mocambique, Naja kaouthia, Perseau americana, Phaseolus coccineus, Phaseolus limensis, Phaseolus vulgaris, Phytolacca americana, Pisum sativum, Pseudomonas aeruginosa, Psophocarpus tetragonolobus, Ptilota plumosa, Ricinus communis, Robinia pseudoacacia, Sambucus nigra, Solanum tuberosum, Sophora japonica, Tetragonolobus purpureas, Triticum vulgaris, Ulex europaeus, Vicia faba, Vicia saliva, Vicia villosa, Vigna radiata, Viscum album, and Wisteria floribunda.

25. The device of claim 19 wherein the first porous separating matrix is impregnated with a carbohydrate capable of aggregating blood cells.

26. The device of claim 25 wherein the carbohydrate is selected from the group consisting of mannitol, sorbitol, inositol, β

-D-glucose, α

-D-glucose, D(+)xylose, D(+)mannose, D(-)arabinose, L(+)arabinose, D(+)galactose, L(-)xylose, D-glucoheptose, L-lyxose, lactose, maltose, and sucrose.

27. The device of claim 26 wherein the carbohydrate is mannitol.

28. The device of claim 19 wherein the second matrix is a membrane for chromatographic separation.

29. The device of claim 28 wherein the membrane for chromatographic separation has a capture zone thereon for binding a member of a specific binding pair.

30. The device of claim 19 wherein the first porous separating matrix is an asymmetric membrane with a first surface and a second surface, the membrane having a gradient of pore sizes such that the pore size decreases from the first surface to the second surface.

31. The device of claim 19 wherein the first porous separating matrix includes two sectors:

(i) a first sector permeable to both the liquid of portion of blood and the cellular components of blood; and

(ii) a second sector permeable to the liquid portion of blood but capable of binding the cellular components of blood.

32. The device of claim 19 wherein the sample application zone includes at least one reagent for treatment of the sample.

33. The device of claim 19 wherein the sample application zone includes a specific binding partner labeled with a detectable label, the specific binding partner having specific binding affinity for at least one component selected from the analyte and a specific binding partner for the analyte in a form that can be resolubilized by the addition of a blood sample to the sample application zone.

34. The device of claim 33 wherein the specific binding partner labeled with a detectable label has specific binding affinity for the analyte.

35. A method for performing an assay for detecting or determining at least one analyte in the liquid portion of a blood sample comprising the steps of:

(a) applying a sample of blood to the porous pad on the first opposable component of the device of claim 33;

(b) allowing the sample to resolubilize the labeled specific binding partner labeled with a detectable label;

(c) bringing the first and second opposable components into opposition to transfer fluid from the first opposable component to the second opposable component by pressure;

(d) allowing the blood sample to flow through the first porous separating matrix of the porous pad to separate the liquid portion of the blood sample from the cellular components of the blood sample;

(e) facilitating the flow of the liquid portion of the blood through the interstices around the trapped cellular components of the blood as a result of the action of the second matrix in the second opposable component; and

(f) allowing the liquid portion of the blood to flow through the second matrix so that an assay is performed in the second matrix, the assay being performed by binding a member of a specific binding pair to the capture zone of the second matrix to detect or determine the at least one analyte, at least one member of the specific binding pair binding to the analyte.