Method of preventing undesired binding in solid phase assays

• US 5,656,504 A
• Filed: 04/25/1995
• Issued: 08/12/1997
• Est. Priority Date: 10/26/1992
• Status: Expired due to Term

• Current Assignee: GE Healthcare Bio-Sciences AB (GE Healthcare Technologies, Inc.)
• Original Assignee: Pharmacia Biosensor AB (General Electric Company)
• Inventors: Johansson, Viveca, Larsson, Anita, Ronnberg, Inger
• Primary Examiner(s): Spiegel, Carol A.
• Application Number:
  US08/424,368
• Time in Patent Office: 840 Days
• Field of Search: 422/82.05, 422/82.06, 422/82.11, 435/7.92-7.95, 435/962, 435/970, 436/518, 436/548, 436/528, 436/529, 436/531, 436/825
• US Class Current: 436/518
• CPC Class Codes: G01N 33/54393 Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
• 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 436/825 Pretreatment for removal of interfering factors from sample

Abstract

In assays of the type which comprise contacting the sample containing the analyte to be detected with a solid phase surface supporting a ligand capable of binding the analyte, undesired binding to the surface is prevented to a substantial degree by adding to the sample one or more components of the material forming the ligand supporting solid phase surface, which components are in at least partially soluble form and capable of interacting with constituents of the sample medium.

Claims

1. A specific binding assay method for determining an analyte in a sample with reduced nonspecific interference from non-analyte constituents in said sample comprising:

(a) providing a solid phase comprising a polymer surface having immobilized thereto a ligand capable of specifically binding said analyte;

(b) adding one or more monomeric, oligomeric or polymeric chain fragments of said polymer to said sample wherein said fragments are at least partially soluble in said sample, to form a reaction medium;

(c) contacting said reaction medium with said solid phase; and

(d) measuring any binding of said analyte to said contacted solid phase to determine the presence or amount of said analyte in said sample, wherein said nonspecific interference is reduced in that said non-analyte constituents bind to said fragments in said reaction medium rather than to said solid phase.

2. The method according to claim 1, wherein said solid phase surface is provided as a carrier matrix coating said solid phase.

3. The method of claim 2, wherein said carrier matrix is a hydrogel.

4. The method according to claim 3, wherein said hydrogel is dextran or a derivative thereof.

5. The method according to claim 4, wherein said dextran or derivative thereof is present at a final concentration up to about 2 mg/ml in said reaction medium.

6. The method according to claim 4, wherein said dextran or derivative thereof is present at a final concentration of about 0.5 to about 1.5 mg/ml in said reaction medium.

7. The method according to claim 3, wherein said hydrogel is a polysaccharide or a water-swellable organic polymer.

8. The method according to claim 3, wherein said polymer and said fragments are polymeric or oligomeric chains of said hydrogel.

9. The method according to claim 8, wherein said polymeric or oligomeric chain is soluble in said reaction medium.

10. The method according to claim 1, wherein said ligand is an antibody and wherein a nonspecific antibody is further added to said reaction medium.

11. The method according to claim 10, wherein said antibody and said nonspecific antibody are mouse monoclonal antibodies.

12. The method according to claim 10, wherein said nonspecific antibody is present at a final concentration up to about 200 μ

g/ml in said reaction medium.

13. The method according to claim 12, wherein said nonspecific antibody is present at a final concentration in the range of from about 5 to about 100 μ

g/ml in said reaction medium.

14. The method according to claim 1, wherein said sample is blood, serum or plasma.

15. The method according to claim 14, wherein said sample is serum or plasma.

16. The method according to claim 1, wherein said measuring step comprises measuring any increase in the mass of said solid phase due to binding thereto of said analyte.

17. The method according to claim 16, wherein said mass measurement is an optical reflection measurement by an evanescent wave sensor.

18. The method according to claim 16, wherein said mass measurement is surface plasmon resonance measurement.

19. The method according to claim 1, further comprising adding an ionic strength increasing agent to said reaction medium in an amount sufficient to inhibit interfering ionic interactions without affecting specific binding between said ligand and said analyte.

20. The method according to claim 19, wherein said ionic strength increasing agent is sodium chloride which is present at a final concentration in the range from about 0.15 to about 0.7M in said reaction mixture.

21. The method according to claim 19, wherein said mass measurement is taken in a flow cell through which a buffer or elution reagent flows and wherein said reaction medium is injected into said flowing buffer or elution agent.

22. The method according to claim 21, wherein said ionic strength agent is provided as part of said flowing buffer or elution reagent.

23. The method according to claim 1, wherein said antibody which is said ligand is an IgG2a or IgG2b antibody.

24. The method according to claim 23, wherein said antibody which is said ligand is a monoclonal antibody.

25. A specific binding assay method for determining an analyte in a sample with reduced nonspecific interference from non-analyte constituents in an aqueous sample containing or suspected of containing the analyte to be detected comprising:

(a) providing a solid phase surface comprising an organic hydrogel surface having immobilized thereto a ligand capable of specifically binding said analyte;

(b) adding one or more water soluble monomeric, oligomeric or polymeric fragments of said organic hydrogel to said sample wherein said fragments are at least partially soluble in said sample, to form a reaction medium;

(c) contacting said reaction medium with said solid phase; and

(d) measuring any binding of said analyte to said contacted solid phase to determine the presence or amount of said analyte in said sample, wherein said nonspecific interference is reduced in that said non-analyte constituents bind to said fragments in said reaction medium rather than to said solid phase.