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

• US 6,008,059 A
• Filed: 08/19/1997
• Issued: 12/28/1999
• 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:
  US08/914,435
• Time in Patent Office: 861 Days
• Field of Search: 422/56, 422/58, 422/61, 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, 435/970, 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: 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;

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

(iii) a conductor in operable contact with the second porous matrix at an end of the second porous matrix opposite that which is in operable contact with the first porous separating matrix; and

(b) a second opposable component attachable to the first opposable component, the second opposable component including thereon;

(i) an applicator; and

(ii) an absorber separated from the applicator;

such that the first and second opposable components are brought into opposition to transfer a reagent, in the applicator, from the second opposable component to the first opposable component by operable contact of the applicator on the second opposable component and the conductor on the first opposable component by pressure so that bringing the first and second opposable components into opposition causes the reagent transferred from the second opposable component to the first opposable component to migrate through the second porous matrix in a second direction opposite to the first direction with flow being driven by the absorber to perform bidirectional immunochromatography;

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 membrane for chromatographic separation includes therein a capture zone for binding an analyte and the reagent transferred from the second opposable component to the first opposable component is a labeled specific binding partner for the analyte.

3. 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 first porous separating matrix on the first opposable component of the device of claim 2;

(b) 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;

(c) allowing the liquid portion of the blood sample to flow through the membrane for chromatographic separation in the first direction;

(d) bringing the first and second opposable components into opposition to transfer the labeled specific binding partner for the analyte from the second opposable component to the first opposable component by pressure by bringing the applicator on the second opposable component into operable contact with the conductor on the first opposable component; and

(e) allowing the labeled specific binding partner for the analyte to flow through the membrane for chromatographic separation in the second direction, the flow driven by the absorber, so that an assay is performed in the second matrix, the assay being performed by binding the labeled specific binding partner to the capture zone of the second matrix to detect or determine the at least one analyte.

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

5. The device of claim 4 wherein the binder is an anti-blood cell antibody.

6. The device of claim 5 wherein the anti-blood cell antibody is an anti-erythrocyte antibody.

7. The device of claim 4 wherein the binder is a lectin.

8. The device of claim 7 where 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.

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

10. The device of claim 9 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.

11. The device of claim 10 wherein the carbohydrate is mannitol.

12. 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.

13. The device of claim 1 wherein the first porous separating membrane includes therein 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 it.

14. 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;

(ii) a second porous separating matrix permeable to the liquid portion of blood but capable of trapping the cellular components of blood, the second porous separating matrix being in operable contact with the first porous separating matrix;

(iii) a third porous matrix including a membrane for chromatographic separation in operable contact with the second porous separating matrix, the third porous matrix permitting the liquid portion of the blood to flow in a first direction by capillary action through the third porous matrix; and

(iv) a conductor in operable contact with the third porous matrix at an end of the third porous matrix opposite that which is in operable contact with the second porous separating matrix; and

(b) a second opposable component attachable to the first opposable component, the second opposable component including thereon;

(i) an applicator; and

(ii) an absorber separated from the applicator;

such that the first and second opposable components can be brought into opposition to transfer a reagent, in the applicator, from the second opposable component to the first opposable component by pressure by operable contact of the applicator on the second opposable component and the conductor on the first opposable component so that bringing the first and second opposable components into opposition causes the reagent transferred from the second opposable component to the first opposable component to migrate through the third porous matrix in a second direction opposite to the first direction with flow being driven by the absorber to perform bidirectional immunochromatography;

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.

15. The device of claim 14 wherein the membrane for chromatographic separation includes therein a capture zone for binding an analyte and the reagent transferred from the second opposable component to the first opposable component includes a labeled specific binding partner for the analyte.

16. The device of claim 14 wherein the either the first porous matrix, the second porous matrix, or both the first and second porous matrices contain a binder for the cellular components of blood.

17. The device of claim 14 wherein the binder is an anti-blood cell antibody.

18. The device of claim 17 wherein the anti-blood cell antibody is an anti-erythrocyte antibody.

19. The device of claim 14 wherein the binder is a lectin.

20. The device of claim 19 wherein a lectin is selected from the group consisting of concanavalin A, abrin, phytohaemagglutinin, limulin, and the lectins produced by the following species:

Agaricus hisporus, 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.

21. The device of claim 14 wherein either the first porous matrix, the second porous matrix, or both the first and second porous matrices are impregnated with a carbohydrate capable of aggregating blood cells.

22. The device of claim 21 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.

23. The device of claim 22 wherein the carbohydrate is mannitol.

24. The device of claim 14 wherein either the first porous matrix, the second porous matrix, or both the first and second porous matrices include 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.

25. The device of claim 14 wherein either the first porous matrix, the second porous matrix, or both the first and second porous matrixes are asymmetric membranes 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 it.

26. 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;

(ii) at least two second porous matrices each including a membrane for chromatographic separation, each second porous matrix in operable conduct with the first porous separating matrix in order to permit the liquid portion of the blood to flow in a first direction by capillary action through the second porous matrices; and

(iii) at least two conductors, each conductor in operable contact with one of the second porous matrices at the end of each of the second porous matrices opposite that which is in operable contact with the first porous separating matrix; and

(b) a second opposable component attachable to the first opposable component such that the first and second opposable components can be brought into opposition to transfer a reagent from the second opposable component to the first opposable component by pressure, the second opposable component including thereon;

(i) at least one applicator; and

(ii) at least one absorber separated from the at least one applicator;

so that bringing the first and second opposable components into opposition causes a reagent, in the at least one applicator, transferred from the second opposable component to the first opposable component to migrate through the second porous matrices in a second direction opposite to the first direction by bringing the at least one applicator on the second opposable component into operable contact with the conductors on the first opposable component, the flow being driven by the at least one absorber to perform bidirectional immunochromatography;

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.

27. The device of claim 26 wherein the membranes for chromatographic separation each include therein a capture zone for binding an analyte and the reagent transferred from the second opposable component to the first opposable component includes a labeled specific binding partner for the analyte.

28. The device of claim 26 wherein the first porous separating matrix contains a binder for the cellular components of blood.

29. The device of claim 28 wherein the binder is an anti-blood cell antibody.

30. The device of claim 29 wherein the anti-blood cell antibody is an anti-erythrocyte antibody.

31. The device of claim 28 wherein the binder is a lectin.

32. The device of claim 31 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.

33. The device of claim 26 wherein the first porous separating matrix is impregnated with a carbohydrate capable of aggregating blood cells.

34. The device of claim 33 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.

35. The device of claim 34 wherein the carbohydrate is mannitol.

36. The device of claim 26 wherein the first 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.

37. The device of claim 26 wherein the first 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 it.