Stabilizing agents and capture ligands for use in assays measuring analyte concentrations

• US 7,897,352 B2
• Filed: 07/20/2007
• Issued: 03/01/2011
• Est. Priority Date: 07/28/2006
• Status: Active Grant

• Current Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Original Assignee: Beckman Coulter Incorporated (Danaher Corporation)
• Inventors: Lu, Wenyuan, Leith, Katherine M., Walton, Courtney E., Chan, Stephen P.
• Primary Examiner(s): Haq; Shafiqul
• Application Number:
  US11/878,121
• Publication Number:
  US 20080182341A1
• Time in Patent Office: 1,320 Days
• Field of Search: None
• US Class Current: 435/7.1
• CPC Class Codes: C07K 16/00 Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies antibodies with enzymatic activity, e.g. abzymes C12N9/0002
• CPC Class Codes: G01N 33/53 Immunoassay; Biospecific binding assay; Materials therefor
• CPC Class Codes: G01N 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
• CPC Class Codes: G01N 33/543 with an insoluble carrier for immobilising immunochemicals
• 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

Abstract

The present invention is related to compositions useful for the measurement of free or unbound analyte concentrations in a fluid. The present invention includes the use of capture ligands and stabilizing agents to improve the accuracy of analyte concentration assays. Methods and tools for using the present invention are also disclosed.

Claims

1. A method for measuring a concentration of a free analyte, wherein analyte exists in an equilibrium between free analyte and bound analyte in a sample, the method comprising:

(a) forming a test mixture by adding to a vessel;

(i) a capture ligand for the free analyte,

(ii) 2-ethyl-hexyl sulfate, or salt thereof in an amount effective to preserve an equilibrium between free analyte and bound analyte, and

(iii) the sample comprising the analyte;

(b) adding a detection system to the vessel; and

(c) measuring the concentration of the analyte in the sample using the detection system.

2. The method of claim 1, wherein the analyte is a thyroid hormone.

3. The method of claim 1, wherein the capture ligand is analyte-specific and selected from the group consisting of antibody, antibody fragment, antibody mimic, analyte-specific binding protein, and combinations thereof.

4. The method of claim 3, wherein the analyte-specific binding protein is intrinsic factor or folate-binding protein.

5. The method of claim 1, wherein the detection system comprises a solid phase coated with a capture ligand-specific binding molecule, a detectable label, or combination thereof.

6. The method of claim 5, wherein the solid phase is a paramagnetic particle.

7. The method of claim 5, wherein the capture ligand-specific binding molecule is hapten-specific, and wherein the capture ligand is haptenated.

8. The method of claim 7, wherein the hapten is biotin.

9. The method of claim 5, wherein the capture ligand-specific binding molecule is selected from the group consisting of streptavidin, avidin, biotin-specific antibody, biotin-specific antibody fragment, and biotin-specific antibody mimic.

10. The method of claim 1, wherein the capture ligand is immobilized on a solid phase.

11. The method of claim 1, wherein a concentration of 2-ethyl-hexyl sulfate in the test mixture is about 5 micromolar to about 750 micromolar.

12. The method of claim 1, wherein a concentration of the 2-ethyl-hexyl sulfate in the test mixture is about 75 micromolar to about 650 micromolar.