Method for capturing analytes eluted from surface-bound ligands

• US 20020012929A1
• Filed: 03/16/2001
• Published: 01/31/2002
• Est. Priority Date: 03/16/2000
• Status: Active Grant

• Current Assignee: GE Healthcare Bio-Sciences AB (General Electric Company)
• Original Assignee: Biacore AB
• Inventors: Malmqvist, Magnus, Jansson, Osten
• Granted Patent: US 6,503,760 B2
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: G01N 21/553 and using surface plasmons fluorescence excitation G01N21/648; enhanced Raman G01N21/658
• CPC Class Codes: G01N 21/648 using evanescent coupling or surface plasmon coupling for the excitation of fluorescence
• CPC Class Codes: G01N 33/537 with separation of immune complex from unbound antigen or antibody
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form

Abstract

Methods for capturing analytes associated with surface-bound ligands are disclosed. The methods involve eluting analytes from surface-bound ligands with a first liquid to generate free analytes, and capturing the free analytes with a solid capturing material within the first liquid to generate a first liquid containing captured analytes. The first liquid may be a flowing liquid or a non-flowing liquid, and the surface to which the surface-bound ligand is attached may be a sensing surface, such as a biosensor, or a non-sensing surface. The captured analytes may be further consolidated at a location removed from the surface-bound ligand, eluted from the solid capturing material with a second liquid, and used for subsequent analysis or procedures.

Claims

1. A method for capturing an analyte associated with a surface-bound ligand, comprising:

eluting the analyte from surface-bound ligand by contacting the surface-bound ligand with a first liquid flow that dissociates the analyte from the surface-bound ligand to generate a free analyte within the first liquid flow; and

capturing the free analyte with a solid capturing material carried within the first liquid flow to generate a first liquid flow containing captured analyte.

2. The method of claim 1 wherein the analyte associated with the surface-bound ligand is an analyte (ligand) interaction pair selected from the group consisting of antigen (antibody), antibody (antigen), hormone (hormone receptor), hormone receptor (hormone), polynucleotide (complementary polynucleotide), avidin/streptavidin (biotin), biotin (avidin/streptavidin), enzyme (enzyme substrate), enzyme (enzyme inhibitor), enzyme substrate (enzyme), enzyme inhibitor (enzyme), lectins (carboxyhydrate), carboxyhydrate (lectins), lipid (lipid binding protein), lipid (membrane-associated protein), lipid binding protein (lipid), membrane-associated protein (lipids), polynucleotide (polynucleotide binding protein), polynucleotide binding protein (polynucleotide), receptor (transmitter), transmitter (receptor), drug (target), target (drug), protein (protein), protein (polynucleotide), polynucleotide (protein), DNA (DNA), DNA (RNA) and RNA (DNA).

3. The method of claim 2 wherein the analyte (ligand) interaction pair is an antigen (antibody) interaction pair.

4. The method of claim 1 wherein the surface to which the surface-bound ligand is bound is a sensing surface.

5. The method of claim 1 wherein the surface to which the surface-bound ligand is bound is a non-sensing surface.

6. The method of claim 4 wherein the sensing surface is a sensing surface of an affinity-based biosensor.

7. The method of claim 6 wherein the affinity-based biosensor is a surface plasmon resonance biosensor.

8. The method of claim 1 wherein the first liquid flow is a laminar flow when in contact with the surface-bound ligand.

9. The method of claim 1 wherein the first liquid flow is an aqueous solution comprising at least one acidic, basic, ionic, organic, detergent or chelating agent.

10. The method of claim 1 wherein the solid capture material are separation beads.

11. The method of claim 10 wherein the separation beads are made from agarose, dextran, hydroxyapatit, silica, polyacrylamid, or hydrophilic polymers.

12. The method of claim 10, wherein the separation beads have magnetic properties.

13. The method of claim 11 wherein the separation beads comprise a chromatographic media having spherical shapes with diameters ranging from 2 to 10 micrometers.

14. The method of claim 1 wherein the steps of eluting and contacting occur within a flow channel of a biosensor.

15. The method of claim 1 wherein a plurality of analytes are associated with a plurality of surface-bound ligands, the step of eluting generates a plurality of free analytes, and the step of capturing generates a plurality of captured analytes.

16. The method of claim 15, further comprising the step of consolidating the plurality of captured analytes at a location removed from the plurality of surface-bound ligands.

17. The method of claim 16 wherein the step of consolidating comprises passing the plurality of captured analytes carried within the first liquid flow through a separation device that prevents passage of the plurality of solid capturing materials having captured analytes associated therewith, while allowing passage of the first liquid flow, and thereby consolidating the captured analytes.

18. The method of claim 17 wherein the separation device comprises a column, and the first liquid flow containing the plurality of captured analytes is passed through the column.

19. The method of claim 17, further comprising eluting the plurality of free analytes from the solid capturing material by contacting the consolidated and captured analytes with a second liquid that dissociates the analytes from the solid capturing material.

20. The method of claim 19 wherein the second liquid is an aqueous solution comprising at least one acidic, basic, ionic, organic, detergent or chelating agent.

21. The method of claim 19 wherein the step of eluting the plurality of free analytes from the solid capturing material occurs in a column employed to consolidate the plurality of captured analytes.

22. The method of claim 19 wherein the plurality of free analytes eluted with the second liquid are collected.

23. The method of claim 22 wherein the collected analytes are subjected to a subsequent analysis.

24. A method for capturing an analyte associated with a surface-bound ligand on a surface of a biosensor, comprising:

eluting the analyte from the surface-bound ligand by contacting the surface-bound ligand with a first liquid that dissociates the analyte from the surface-bound ligand to generate a free analyte within the first liquid; and

capturing the free analyte with a solid capturing material within the first liquid to generate a first liquid containing captured analyte.

25. The method of claim 24 wherein the analyte associated with the surface-bound ligand is an analyte (ligand) interaction pair selected from the group consisting of antigen (antibody), antibody (antigen), hormone (hormone receptor), hormone receptor (hormone), polynucleotide (complementary polynucleotide), avidin/streptavidin (biotin), biotin (avidin/streptavidin), enzyme (enzyme substrate), enzyme (enzyme inhibitor), enzyme substrate (enzyme), enzyme inhibitor (enzyme), lectins (carboxyhydrate), carboxyhydrate (lectins), lipid (lipid binding protein), lipid (membrane-associated protein), lipid binding protein (lipid), membrane-associated protein (lipids), polynucleotide (polynucleotide binding protein), polynucleotide binding protein (polynucleotide), receptor (transmitter), transmitter (receptor), drug (target), target (drug), protein (protein), protein (polynucleotide), polynucleotide (protein), DNA (DNA), DNA (RNA) and RNA (DNA).

26. The method of claim 25 wherein the analyte (ligand) interaction pair is an antigen (antibody) interaction pair.

27. The method of claim 24 wherein the biosensor is an affinity-based biosensor.

28. The method of claim 27 wherein the affinity-based biosensor is a surface plasmon resonance biosensor.

29. The method of claim 24 wherein the first liquid is an aqueous solution comprising at least one acidic, basic, ionic, organic, detergent or chelating agent.

30. The method of claim 24 wherein the solid capture material are separation beads.

31. The method of claim 30 wherein the separation beads are made from agarose, dextran, hydroxyapatit, silica, polyacrylamid, or hydrophilic polymers.

32. The method of claim 30 wherein the separation beads have magnetic properties.

33. The method of claim 31 wherein the separation beads comprise chromatographic media having spherical shapes with diameters ranging from 2 to 10 micrometers.

34. The method of claim 24 wherein a plurality of analytes are associated with a plurality of surface-bound ligands, the step of eluting generates a plurality of free analytes, and the step of capturing generates a plurality of captured analytes.

35. The method of claim 34, further comprising the step of consolidating the plurality of captured analytes at a location removed from the plurality of surface-bound ligands.

36. The method of claim 35 wherein the step of consolidating comprises passing the plurality of captured analytes carried within the first liquid through a separation device that prevents passage of the plurality of solid capturing materials having captured analytes associated therewith, while allowing passage of the first liquid flow, and thereby consolidating the captured analytes.

37. The method of claim 36 wherein the separation device comprises a column, and the first liquid containing the plurality of captured analytes is passed through the column.

38. The method of claim 36, further comprising eluting the plurality of free analytes from the solid capturing material by contacting the consolidated and captured analytes with a second liquid that dissociates the analytes from the solid capturing material.

39. The method of claim 38 wherein the second liquid is an aqueous solution comprising at least one acidic, basic, ionic, organic, detergent or chelating agent.

40. The method of claim 38 wherein the step of eluting the plurality of free analytes from the solid capturing material occurs in a column employed to consolidate the plurality of captured analytes.

41. The method of claim 38 wherein the plurality of free analytes eluted with the second liquid are collected.

42. The method of claim 41 wherein the collected analytes are subjected to a subsequent analysis.