Binding assays using more than one label for determining analyte in the presence of interfering factors

• US 6,087,088 A
• Filed: 01/31/1997
• Issued: 07/11/2000
• Est. Priority Date: 01/31/1997
• Status: Expired due to Term

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Bayer Corporation (Bayer AG)
• Inventors: Piran, Uri, Quinn, John J.
• Primary Examiner(s): GITOMER, RALPH J
• Application Number:
  US08/791,591
• Time in Patent Office: 1,257 Days
• Field of Search: 435/4, 435/7.92, 435/7.94, 435/7.93, 435/7.9, 435/962, 436/518, 436/538, 436/540
• US Class Current: 435/4
• CPC Class Codes: G01N 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/54326 Magnetic particles

Abstract

Novel binding assay techniques have been developed which improve accuracy and sensitivity via accounting for interfering factors. They rely on use, in a simultaneous incubation, of two or more different labels, some of which are used primarily to detect analyte, and others to detect interfering substances originating in the sample. The mathematical relationships between the labels allow corrections that lead to more accurate and sensitive determination of the presence and concentration of the analyte.

Claims

1. A binding assay for determining an amount of an analyte in a sample containing interfering factors comprising

a. contacting said sample containing said analyte with

(1) a labeled specific binder, labeled with a first label, said labeled specific binder binding to said analyte to form an analyte-labeled specific binder complex,

(2) a solid phase containing an immobilized analyte derivative or mimic, said analyte derivative or mimic binding to said labeled specific binder, and

(3) a labeled reference binder, labeled with a second label, which binds to said analyte derivative or mimic and said interfering factors but which does not bind to said analyte,

such that said first label is different from said second label,

b. separating said first and second labels which are bound to said solid phase from said unbound labels,

c. determining the amount of said first and second labels which are bound to said solid phase,

d. comparing the amount of said bound first label with that of reference standards to determine the amount of analyte present , and

e. correcting the amount of said analyte present by (1) comparing said amount of said bound second label with that of reference standards to calculate a correction factor, and (2) applying said correction factor to the amount of analyte determined in step d to correct the amount of said analyte.

2. The binding assay of claim 1 in which said analyte is an immunochemical component, genetic material or a receptor.

3. The binding assay of claim 1 in which there are 3 or more labels, each being detectable in the presence of the others, and each being bound to a different analyte or interference in the assay sample.

4. The binding assay of claim 3 in which 2 or more of the labels are affected by different interfering factors.

5. The binding assay of claim 3 in which 2 or more of the labels are affected by different analytes.

6. The binding assay of claim 1 in which said interfering factor increases the signal of said first or second labels.

7. The binding assay of claim 1 in which said interfering factor decreases the signal of said first or second labels.

8. The binding assay of claim 1 in which said labels are chemiluminescent molecules, acridinium esters or derivatives thereof.

9. The binding assay of claim 1 in which said analyte is selected from the group consisting of proteins, peptides, drugs, hormones, receptors, antibodies, microorganisms, environmental pollutants, nucleic acids, lipids, surfactants, carbohydrates, and conjugates of said analytes.

10. The binding assay of claim 1 in which said analyte is selected from the group consisting of whole cells and cell fragments.

11. The binding assay of claim 1 in which said first label is dimethyl acridinium ester, said second label is a long-emission acridinium ester and said solid phase is a paramagnetic particle.

12. The binding assay of claim 1 in which the presence of an interfering factor is qualitatively detected, wherein said step e comprises detecting the presence of said interfering factor by comparing the amount of said second bound label with that of reference standards.

13. A binding assay for determining an amount of an analyte in a sample containing interfering factors comprising

a. contacting said sample containing said analyte with

(1) a labeled specific binder, labeled with a first label, said labeled specific binder binding to said analyte to form an analyte-labeled specific binder complex,

(2) a solid phase containing an immobilized specific binder, said specific binder binding to said analyte, and

(3) a labeled reference binder, labeled with a second label, which binds to said immobilized specific binder and said interfering factors but which does not bind to analyte,

such that said first label is different from said second label,

b. separating said first and second labels which are bound to said solid phase from said unbound labels,

c. determining the amount of said first and second labels which are bound to said solid phase,

d. comparing the amount of said bound first label with that of reference standards to determine the amount of analyte present , and

e. correcting the amount of said analyte present by (1) comparing said amount of said bound second label with that of reference standards to calculate a correction factor, and

(2) applying said correction factor to the amount of analyte determined in step d to correct the amount of said analyte.

14. The binding assay of claim 13 in which said analyte is an antigen, antibody, genetic material or a receptor.

15. The binding assay of claim 13 in which said first label is dimethyl acridinium ester, said second label is a long-emission acridinium ester and said solid phase is a paramagnetic particle.

16. The binding assay of claim 13 in which there are 3 or more labels, each being detectable in the presence of the others, and each being bound to a different analyte or interfering factor in the assay sample.

17. The binding assay of claim 16 in which 2 or more of the labels are bound to different interfering factors.

18. The binding assay of claim 16 in which 2 or more of the labels are bound to different analytes.

19. The binding assay of claim 13 in which the presence of an interfering factor is qualitatively detected, wherein said step e comprises detecting the presence of said interfering factor by comparing the amount of said second bound label with that of reference standards.

20. The binding assay of claim 13 in which said interfering factors increase the signal of said first or second labels.

21. The binding assay of claim 13 in which said interfering factors decrease the signal of said first or second labels.

22. The binding assay of claim 13 in which said labels are chemiluminescent molecules, acridinium esters or derivatives thereof.

23. The binding assay of claim 13 in which said analyte is selected from the group consisting of proteins, peptides, drugs, hormones, receptors, antibodies, microorganisms, environmental pollutants, nucleic acids, lipids, surfactants, carbohydrates, and conjugates of said analytes.

24. The binding assay of claim 13 in which said analyte is selected from the group consisting of whole cells and cell fragments.