Homogeneous process for the detection and/or determination by luminescence of an analyte in a medium in which it may be present

• US 5,279,943 A
• Filed: 01/19/1993
• Issued: 01/18/1994
• Est. Priority Date: 08/02/1985
• Status: Expired due to Term

• Current Assignee: Compagnie Oris Industrie
• Original Assignee: Compagnie Oris Industrie
• Inventors: Mathis, Gerard, Davin, Thierry
• Primary Examiner(s): WARDEN, JILL ALICE
• Application Number:
  US08/005,610
• Time in Patent Office: 364 Days
• Field of Search: 436/525, 436/537, 436/546, 436/800, 436/808, 436/813, 436/817, 436/805, 436/811, 436/536, 422/61, 435/7.32, 435/7.23, 435/7.4, 435/7.5, 435/968, 435/975
• US Class Current: 435/7.32
• CPC Class Codes: G01N 33/542 with steric inhibition or signal modification, e.g. fluorescent quenching
• CPC Class Codes: Y10S 435/968 High energy substrates, e.g. fluorescent, chemiluminescent, radioactive
• CPC Class Codes: Y10S 435/975 Kit
• CPC Class Codes: Y10S 436/811 Test for named disease, body condition or organ function
• CPC Class Codes: Y10S 436/813 Cancer
• CPC Class Codes: Y10S 436/817 Steroids or hormones

Abstract

The present invention relates to a homogeneous process for the detection and/or determination of an analyte in a medium in which it may be present, by disclosing the reaction product of the analyte and a corresponding receptor, process consisting in: 1) adding to said medium a first reagent consisting of a receptor for the said analyte; 2) adding a second reagent consisting of at least one of the components of the reaction product of the analyte and at least one of its receptors; one of the two reagents being coupled with a luminescent compound and the other reagent possessing a heavy atom or units containing a heavy atom; 3) incubating the medium after addition of each reagent or after the addition of both reagents; 4) exciting the resulting medium and 5) measuring at equilibrium or during the kinetics, the signal emitted by the luminescent compound, said signal being modulated by the heavy atom effect.

Claims

1. A homogeneous process for the detection or determination of an analyte in a medium in which it may be present by disclosing the reaction product of the analyte and a corresponding receptor comprising the steps of

a) adding to said medium a first reagent consisting of a receptor for said analyte;

b) optionally incubating said medium after addition of said first reagent;

c) adding to said medium a second reagent consisting of at least one of said analyte and said receptor for analyte, one of said first and second reagents being coupled with a luminescent compound and the other of said first and second reagents possessing a heavy atom or units containing a heavy atom;

d) incubating said medium after addition of said second reagent to produce a resulting medium;

e) exciting said resulting medium to produce luminescence;

f) measuring said luminescence at equilibrium or under kinetic conditions, wherein said heavy atom is capable of modulating said luminescence in a medium in which analyte is present, wherein said modulation is by inter-system transition within said luminescent compound and wherein there is substantially no overlap between an absorption spectrum of said heavy atom and an emission spectrum of said luminescent compound, and

g) comparing said luminescence to a standard in order to determine the presence or concentration of analyte in said medium.

2. The process of claim 1 characterized by being an excess process of determination or detection wherein said first reagent is coupled with a luminescent compound and wherein said second reagent consists of at least one additional receptor for said analyte and possesses a heavy atom or units containing a heavy atom.

3. The process of claim 1 characterized by being a competition process of determination or detection wherein said first reagent possesses a heavy atom or units containing a heavy atom and wherein said second reagent consists of said analyte coupled with a luminescent compound.

4. The process of claim 1 characterized by being a competition process of determination or detection wherein said first reagent is coupled with a luminescent compound and wherein said second reagent consists of said analyte and possesses a heavy atom or units containing a heavy atom.

5. The process according to claim 1, 2, 3, or 4 wherein the analyte is selected from the group consisting of antibodies, antigens, toxins, enzymes, proteins, hormones, steroids, avidin, biotin, micro-organisms and haptens.

6. The process according to claim 1, 2, 3 or 4 wherein the analyte is a drug.

7. The process according to claim 1, 2, 3, or 4 wherein the analyte is prolactin or a carcinoembryonic antigen.

8. The process according to claim 1, 2, 3, or 4 wherein the luminescent compound is a fluorescent, chemiluminiscent, or phosphorescent compound.

9. The process according to claim 5 wherein the luminescent compound is a fluorescent, chemiluminescent, or phosphorescent compound.

10. The process according to claim 7 wherein the luminescent compound is a fluorescent compound selected from the group consisting of fluorescein, rare earth cryptates and chelates.

11. The process according to claim 1, 2, 3, or 4 wherein one of said first and second reagents is labelled with fluorescein and the other of said first and second reagents is iodinated.

12. The process according to claim 1, 2, 3, or 4 wherein the luminescent compound has a luminescent decay lifetime longer than said medium and wherein the excitation of the resulting medium is a pulsed excitation.

13. The process according to claim 5 wherein the luminescent compound is a rare earth cryptate.

14. The process according to claim 2 for the determination or detection of an analyte in a medium, wherein said analyte is an antigen or a hapten and wherein said first reagent is a fluorescein-labelled first antibody for said antigen or hapten and said second reagent is an iodinated second antibody, in less than excess amount, for said antigen or hapten, said first and second antibodies having different specificities for the antigen or hapten.

15. The process according to claim 2 for the determination or detection of an analyte in a medium, wherein said analyte is an antigen or a hapten and wherein said first reagent is a fluorescein-labelled first antibody, in less than excess amount, for said antigen or hapten and said second reagent is an iodinated second antibody for said antigen or hapten, said first and second antibodies having different specificities for the antigen or hapten.

16. The process according to claim 3 for the determination or detection of an analyte in a medium, wherein said analyte is an antigen or a hapten and wherein said first reagent is an iodinated antibody for said antigen or hapten and said second reagent is the antigen or hapten, in a given quantity, labelled with fluorescein.

17. The process according to claim 4 for the determination or detection of an analyte in a medium, wherein said analyte is an antigen or a hapten and wherein said first reagent is a fluorescein-labelled antibody for said antigen or hapten and said second reagent is the iodinated antigen or hapten in a given quantity.

18. The process according to claim 14, 15, 16, or 17 wherein the light excitation of the resulting medium is a pulsed excitation.

19. A kit for the homogeneous detection or determination in liquid phase of an analyte in a medium in which it may be present by measuring luminescence of a luminescent compound, the kit comprising

a) a first reagent consisting of at least one receptor for the analyte to be determined;

b) a second reagent consisting of at least one of said analyte and said receptor for analyte, one of said first and second reagents being coupled with said luminescent compound and the other of first and second reagents possessing a heavy atom or units containing at least one heavy atom, said heavy atom being capable of modulating the light emitted by said luminescent compound upon excitation, said modulation being by inter-system transition within said luminescent compound and wherein there is substantially no overlap between an absorption spectrum of said heavy atom and an emission spectrum of said luminescent compound; and

c) standard samples containing known quantities of the analyte to be determined, for establishing standard curves.

20. The kit of claim 19 further comprising a diluent and a buffer.

21. The kit according to claim 19 wherein the luminescent compound is a chemiluminescent compound and wherein the kit further comprises chemical reagents capable of producing light excitation.

22. The kit of claim 19, 20, or 21 for the determination or detection of prolactin or a carcinoembryonic antigen, wherein said standard samples contain known quantities of prolactin or a carcinoembryonic antigen.