Method and apparatus for improved luminescence assays using particle concentration chemiluminescence detection

• US 6,448,091 B1
• Filed: 11/07/1994
• Issued: 09/10/2002
• Est. Priority Date: 11/03/1988
• Status: Expired due to Fees

• Current Assignee: BioVeris Corporation (Roche Holding AG)
• Original Assignee: IGEN International, Inc. (Roche Holding AG)
• Inventors: Shah, Haresh P., Blackburn, Gary F., Wilkins, Elizabeth W., Massey, Richard J.
• Primary Examiner(s): Venkat, Jyothsna
• Assistant Examiner(s): FRIEND, TOMAS H F
• Application Number:
  US08/335,183
• Time in Patent Office: 2,864 Days
• Field of Search: 435/7.1, 435/7.7, 435/7.72, 435/7.9, 435/7.91, 435/7.92, 435/25, 435/28, 435/968, 436/518, 436/526, 436/805
• US Class Current: 436/501
• CPC Class Codes: C07H 21/00 Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids
• CPC Class Codes: C12Q 1/6816 characterised by the detection means C12Q1/6804 takes precedence
• CPC Class Codes: C12Q 1/6825 Nucleic acid detection involving sensors
• CPC Class Codes: C12Q 2563/103 luminescence
• CPC Class Codes: C12Q 2563/131 the label being a member of a cognate binding pair, i.e. extends to antibodies, haptens, avidin
• CPC Class Codes: C12Q 2565/102 Multiple non-interacting labels
• CPC Class Codes: G01N 1/40 Concentrating samples
• CPC Class Codes: G01N 21/69 specially adapted for fluids , e.g. molten metal
• CPC Class Codes: G01N 21/76 Chemiluminescence; Bioluminescence
• CPC Class Codes: G01N 2458/30 Electrochemically active labels
• CPC Class Codes: G01N 33/54313 the carrier being characterised by its particulate form
• CPC Class Codes: G01N 33/54326 Magnetic particles
• CPC Class Codes: G01N 33/54366 Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/5438 Electrodes
• CPC Class Codes: G01N 33/582 with fluorescent label
• CPC Class Codes: G01N 33/585 with a particulate label, e.g. coloured latex

Abstract

Methods and apparatus for performing a binding assay for an analyte of interest present in a sample are described. The methods include the steps of: forming a composition containing the sample, an assay-performance-substance which contains a component linked to a label compound capable of chemiluminescing when triggered, and a plurality of particles capable of specifically binding with the analyte and/or the assay-performance-substance; incubating the composition to form a complex which includes a particle and the labeled component; collecting the complex in a collection zone; introducing into the collection zone a trigger capable of triggering the label such that the label luminesces; and measuring the emitted luminescence to measure the presence of the analyte of interest in the sample.

Claims

1. A method for performing a binding assay for an analyte of interest possibly present in a sample comprising:

(a) forming a composition containing;

(i) said sample;

(ii) an assay-performance-substance comprising at least one compound selected from the group consisting of;

(1) added analyte of interest or added analogue of said analyte;

(2) a binding partner of said analyte or a binding partner of said analogue; and

(3) a component capable of binding with (1) or (2);

wherein said assay-performance-substance further contains a label compound linked to at least one of said (1), (2), and (3) and wherein said label compound is capable of chemiluminescing when triggered; and

(iii) a plurality of particles capable of specifically binding with the analyte and/or said assay-performance-substance;

(b) incubating said composition to form a complex which includes a particle and said labeled component;

(c) introducing a suspension of said particles into a flow-cell;

(d) collecting said particles in a collection zone contained within said flow-cell thereby removing said particles from suspension, wherein said composition is flowed through the flow-cell at a sufficiently low rate of flow to permit collection of at least a portion of said particles;

(e) introducing into said collection zone a trigger capable of triggering said label such that said label chemiluminesces; and

(f) measuring the emitted chemiluminescence from said label on said complex while said particles are collected at said collection zone to measure the presence of the analyte of interest in the sample.

2. A method as recited in claim 1 wherein said trigger is an oxidant capable of oxidizing said label.

3. A method as recited in claim 1 wherein said particles are magnetically responsive and said complex is magnetically collected in said collection zone.

4. A method as recited in claim 2 wherein said oxidant is hydrogen peroxide or superoxide.

5. A method as recited in claim 1 wherein said assay-performance-substance further contains an enzyme for converting a precursor to an oxidant capable of oxidizing said label, and said trigger is the precursor.

6. A method as recited in claim 5 wherein the enzyme is glucose oxidase, the precursor is glucose and the oxidant is hydrogen peroxide.

7. A method as recited in claim 1 wherein said particles further contain an enzyme for converting a precursor to an oxidant capable of oxidizing said label, and said trigger is the precursor.

8. A method as recited in claim 7 wherein the enzyme is glucose oxidase, the precursor is glucose and the oxidant is hydrogen peroxide.

9. A method as recited in claim 1 conducted as a batch process, the composition being permitted to reside within said cell for a time sufficient to permit collection of said particles.

10. A method as recited in claim 7 wherein said particles have a density of from 0.1 to 5 g/mL.

11. A method as recited in claim 10 wherein said particles have a density of from 0.5 to 2 g/mL.

12. A method as recited in claim 7 wherein the size of said particles, measured as the mean diameter, ranges from 0.001 to 100 μ

m.

13. A method as recited in claim 12 wherein the size of said particles ranges from 0.01 to 10 μ

m.

14. An assay method as recited in claim 7 wherein the concentration of particles in said composition is from 1 to 10,000 μ

g/mL.

15. A method as recited in claim 14 wherein said concentration of particles is in the range of from 5 to 1000 μ

g/mL.

16. A method as recited in claim 7 wherein said particles have a magnetic susceptibility of at least 0.001 cgs units.

17. A method as recited in claim 16 wherein the magnetic susceptibility is at least 0.01 cgs units.

18. A method as recited in claim 7 wherein the magnetic susceptibility, density, size and concentration of said particles in said composition is such that the settling rate of said particles is at least 0.5 mm/min.

19. A method as recited in claim 1, wherein said complex consists essentially of said analyte, said particle and said labeled component.

22. The method of claim 1, wherein said method does not involve separating said plurality of particles from free-label compound prior to introducing said suspension of particles into said flow-cell.

24. The method of claim 1, wherein said collection zone comprises a magnet.

25. The method of claim 1, wherein said collection is performed magnetically.

20. A method for performing a binding assay for an analyte of interest possibly present in a sample comprising:

(a) forming a composition containing;

(i) said sample;

(ii) an assay-performance-substance comprising at least one compound selected from the group consisting of;

(1) added analyte of interest or added analogue of said analyte;

(2) a binding partner of said analyte or a binding partner of said analogue; and

(3) a component capable of binding with (1) or (2);

wherein said assay-performance-substance further contains a label compound linked to at least one of said (1), (2), and (3) and wherein said label compound is capable of chemiluminescing when triggered; and

(iii) a plurality of particles capable of specifically binding with the analyte and/or said assay-performance-substance;

(b) incubating said composition to form a complex which includes a particle and said labeled component;

(c) introducing a suspension of said particles into a cell;

(d) collecting said particles in a collection zone within said cell thereby removing said particles from suspension;

(e) introducing into said collection zone a trigger capable of triggering said label such that said label chemiluminesces; and

(f) measuring the emitted chemiluminescence from said label on said complex while said particles are collected at said collection zone to measure the presence of the analyte of interest in the sample.

23. The method of claim 20, wherein said method does not involve separating said plurality of particles from free-label compound prior to introducing said suspension of particles into said cell.

26. The method of claim 20, wherein said collection zone is in a flow-through cell.

21. A method for performing a binding assay for an analyte of interest possibly present in a sample consisting essentially of:

(a) forming a composition containing;

(i) said sample;

(ii) an assay-performance-substance comprising at least one compound selected from the group consisting of;

(1) added analyte of interest or added analogue of said analyte;

(2) a binding partner of said analyte or a binding partner of said analogue; and

(3) a component capable of binding with (1) or (2);

wherein said assay-performance-substance further contains a label compound linked to at least one of said (1), (2), and (3) and wherein said label compound is capable of chemiluminescing when triggered; and

(iii) a plurality of particles capable of specifically binding with the analyte and/or said assay-performance-substance;

(b) incubating said composition to form a complex which includes a particle and said labeled component;

(c) introducing a suspension of said particles into a cell;

(d) collecting said particles in a collection zone within said cell thereby removing said particles from suspension;

(e) introducing into said collection zone a trigger capable of triggering said label such that said label chemiluminesces; and

(f) measuring the emitted chemiluminescence from said label on said complex while said particles are collected at said collection zone to measure the presence of the analyte of interest in the sample.

27. The method of claim 21, wherein said collection zone is in a flow-through cell.

28. A method for performing a binding assay for an analyte of interest possibly present in a sample consisting essentially of:

(a) forming a composition containing;

(i) said sample;

(ii) an assay-performance-substance comprising at least one compound selected from the group consisting of;

(1) added analyte of interest or added analogue of said analyte;

(2) a binding partner of said analyte or a binding partner of said analogue; and

(3) a component capable of binding with (1) or (2);

wherein said assay-performance-substance further contains a label compound linked to at least one of said (1), (2), and (3) and wherein said label compound is capable of chemiluminescing when triggered; and

(iii) a plurality of particles capable of specifically binding with the analyte and/or said assay-performance-substance;

(b) incubating said composition to form a complex which includes a particle and said labeled component;

(c) introducing a suspension of said particles into a cell;

(d) collecting said particles in a collection zone within said cell thereby removing said particles from suspension;

(e) flowing a fluid through said cell thereby separating unbound components from said particles;

(f) introducing into said collection zone a trigger capable of triggering said label such that said label chemiluminesces; and

(g) measuring the emitted chemiluminescence from said label on said complex while said particles are collected at said collection zone to measure the presence of the analyte of interest in the sample.

29. A method for performing a binding assay for an analyte of interest possibly present in a sample comprising:

(a) forming a suspension in a cell, said suspension containing;

(i) said sample;

(ii) an assay-performance-substance comprising at least one compound selected from the group consisting of;

(1) added analyte of interest or added analogue of said analyte;

(2) a binding partner of said analyte or a binding partner of said analogue; and

(3) a component capable of binding with (1) or (2);

wherein said assay-performance-substance further contains a label compound linked to at least one of said (1), (2), and (3) and wherein said label compound is capable of chemiluminescing when triggered; and

(iii) a plurality of particles capable of specifically binding with the analyte and/or said assay-performance-substance;

(b) collecting said particles in a collection zone within said cell thereby removing said particles from suspension;

(c) flowing a fluid through said cell thereby separating unbound components from said particles;

(d) introducing into said collection zone a trigger capable of triggering said label such that said label chemiluminesces; and

(e) measuring the emitted chemiluminescence from said label on said complex while said particles are collected at said collection zone to measure the presence of the analyte of interest in the sample.

30. A method for performing a binding assay for an analyte of interest possibly present in a sample consisting essentially of:

(a) forming a suspension in a cell, said suspension containing;

(i) said sample;

(ii) an assay-performance-substance comprising at least one compound selected from the group consisting of;

(1) added analyte of interest or added analogue of said analyte;

(2) a binding partner of said analyte or a binding partner of said analogue; and

(3) a component capable of binding with (1) or (2);

wherein said assay-performance-substance further contains a label compound linked to at least one of said (1), (2), and (3) and wherein said label compound is capable of chemiluminescing when triggered; and

(iii) a plurality of particles capable of specifically binding with the analyte and/or said assay-performance-substance;

(b) collecting said particles in a collection zone within said cell thereby removing said particles from suspension;

(c) flowing a fluid through said cell thereby separating unbound components from said particles;

(d) introducing into said collection zone a trigger capable of triggering said label such that said label chemiluminesces; and

(e) measuring the emitted chemiluminescence from said label on said complex while said particles are collected at said collection zone to measure the presence of the analyte of interest in the sample.