ASSAYS FOR HUMAN NT-PRO B-TYPE NATRIURETIC PEPTIDE, HUMAN PRO B-TYPE NATRIURETIC PEPTIDE AND HUMAN B-TYPE NATRIURETIC PEPTIDE

• US 20110124012A1
• Filed: 11/21/2009
• Published: 05/26/2011
• Est. Priority Date: 11/21/2009
• Status: Active Grant

• Current Assignee: Abbott Laboratories Incorporated
• Original Assignee: Abbott Laboratories Incorporated
• Inventors: Moore, Jeffrey Allen, Konrath, John George
• Granted Patent: US 8,455,212 B2
• Time in Patent Office: Days
• US Class Current: 435/7.9
• CPC Class Codes: G01N 2333/58 Atrial natriuretic factor complex; Atriopeptin; Atrial natriuretic peptide [ANP]; Brain natriuretic peptide [BNP, proBNP]; Cardionatrin; Cardiodilatin
• CPC Class Codes: G01N 33/6887 from muscle, cartilage or connective tissue
• CPC Class Codes: G01N 33/74 involving hormones or other non-cytokine intercellular protein regulatory factors such as growth factors, including receptors to hormones and growth factors

Abstract

The present disclosure relates to assays for detecting and/or quantifying the amount of human NT-pro B-type natriuretic peptide, human pro B-type natriuretic peptide and human B-type natriuretic peptide in a test sample.

Claims

1. An immunoassay for quantifying the amount of human NT-pro B-type natriuretic peptide (“

human NT-proBNP”

), human pro B-type natriuretic peptide (“

human proBNP”

) and human brain natriuretic peptide (“

hBNP”

) in a test sample being tested for or suspected of containing human NT-proBNP, human proBNP and hBNP, the method comprising the steps of;

(a) contacting a test sample with (i) a first capture antibody that binds to human NT-proBNP and that has been immobilized onto a solid phase to produce a first immobilized antibody and forming a first mixture comprising a first capture antibody-human NT-proBNP complex;

(ii) a second capture antibody that binds to human proBNP and that has been immobilized onto a solid phase to produce a second immobilized antibody and forming a second mixture comprising a second capture antibody-human proBNP complex; and

(iii) the second capture antibody, which in addition to binding to human proBNP also binds hBNP, and that has been immobilized onto a solid phase to produce a third immobilized antibody and forming a third mixture comprising a second capture antibody-hBNP complex, wherein said first capture antibody comprises antibody 15F11 and the second capture antibody comprises antibody 8.1;

(b) contacting said first mixture comprising the first capture antibody-human NT-proBNP complex with a first detection antibody which binds to human NT-proBNP and that has been conjugated to a detectable label to form a third mixture comprising a first capture antibody-human NT-proBNP-first detection antibody complex, wherein the first detection antibody antibody 15C4;

(c) contacting the second mixture comprising the second capture antibody-human proBNP complex with the first detection antibody which, in addition to binding to human NT-proBNP, also binds to human proBNP, and that has been conjugated to a detectable label to form a fifth mixture comprising a second capture antibody-human proBNP-first detection antibody complex, wherein the first detection antibody is antibody 15C4;

(d) contacting the third mixture comprising the second capture antibody-hBNP complex with a second detection antibody which binds hBNP and that has been conjugated to a detectable label to form a sixth mixture comprising a second capture antibody-hBNP-second detection antibody complex, wherein the second detection antibody is antibody 201.3; and

(e) determining the amount of (i) first capture antibody-human NT-proBNP-first detection antibody complex formed in step (b) by detecting the detectable label as a measure of the amount of human NT-proBNP contained in the test sample;

(ii) the second capture antibody-human proBNP-first detection antibody complex formed in step (c) by detecting the detectable label as a measure of the amount of human proBNP contained in the test sample; and

(iii) the second capture antibody-hBNP-second detection antibody complex formed in step (d) by detecting the detectable label as a measure of the amount of hBNP contained in the test sample.

2. The immunoassay of claim 1, wherein the solid phase is selected from the group consisting of a magnetic particle, bead, test tube, microtiter plate, cuvette, membrane, a scaffolding molecule, film, filter paper, disc, and chip.

3. The immunoassay of claim 1, wherein the detectable label of the first detection antibody is selected from the group consisting of a radioactive label, an enzymatic label, a chemiluminescent label, a fluorescent label, a thermometric label, and an immuno-polymerase chain reaction label.

4. The immunoassay of claim 1, wherein the detectable label of the second detection antibody is selected from the group consisting of a radioactive label, an enzymatic label, a chemiluminescent label, a fluorescent label, a thermometric label, and an immuno-polymerase chain reaction label.

5. An immunoassay for quantifying the amount of human NT-pro B-type natriuretic peptide (“

human NT-proBNP”

), human pro B-type natriuretic peptide (“

human proBNP”

) and human brain natriuretic peptide (“

hBNP”

) in a test sample being tested for or suspected of containing human NT-proBNP, human proBNP and hBNP, the method comprising the steps of;

(a) contacting a test sample with (i) a first detection antibody which binds to human NT-proBNP and that has been conjugated to a detectable label to form a first mixture comprising a human NT-proBNP-first detection antibody complex;

(ii) the first detection antibody, which in addition to binding to human NT-proBNP also binds to human proBNP, and that has been conjugated to a detectable label to form a second mixture comprising a human proBNP-first detection complex; and

(iii) a second detection antibody which binds to hBNP and that has been conjugated to a detectable label to further form in the third mixture comprising a hBNP-second detection antibody complex, wherein the first detection antibody is antibody 15C4 and the second detection antibody is antibody 201.3;

(b) contacting said first mixture comprising the human NT-proBNP-first detection antibody complex with a first capture antibody that binds to human NT-proBNP and that has been immobilized onto a solid phase to produce a first immobilized antibody to form a fourth mixture comprising a first capture antibody-human NT-proBNP-first detection antibody complex, wherein the first capture antibody is 15F11;

(c) contacting said second mixture comprising the human proBNP-first detection antibody complex with a second capture antibody that binds to human proBNP and that has been immobilized onto a solid phase to produce a second immobilized antibody to form a fifth mixture comprising a second capture antibody-human proBNP-first detection antibody complex, wherein the second capture antibody is 8.1;

(d) contacting the third mixture comprising the hBNP-second detection antibody complex with the second capture antibody, which in addition to binding to human proBNP also binds to hBNP, and that has been immobilized onto a solid phase to produce a second immobilized antibody to form a sixth mixture comprising a second capture antibody-human BNP-second detection antibody complex, wherein the second capture antibody is 8.1;

(d) determining the amount of (i) first capture antibody-human NT-proBNP-first detection antibody complex formed in step (b) by detecting the detectable label as a measure of the amount of human NT-proBNP contained in the test sample;

(ii) the second capture antibody-human proBNP-first detection antibody complex formed in step (c) by detecting the detectable label as a measure of the amount of human proBNP contained in the test sample; and

(iii) the second capture antibody-hBNP-second detection antibody complex formed in step (d) by detecting the detectable label as a measure of the amount of hBNP contained in the test sample.

6. The immunoassay of claim 5, wherein the solid phase is selected from the group consisting of a magnetic particle, bead, test tube, microtiter plate, cuvette, membrane, a scaffolding molecule, film, filter paper, disc, and chip.

7. The immunoassay of claim 5, wherein the detectable label of the first detection antibody is selected from the group consisting of a radioactive label, an enzymatic label, a chemiluminescent label, a fluorescent label, a thermometric label, and an immuno-polymerase chain reaction label.

8. The immunoassay of claim 5, wherein the detectable label of the second detection antibody is selected from the group consisting of a radioactive label, an enzymatic label, a chemiluminescent label, a fluorescent label, a thermometric label, and an immuno-polymerase chain reaction label.

9. Immunodiagnostic reagents comprising:

(a) a first capture antibody specific for human NT-pro B-type natriuretic peptide (“

human NT-proBNP”

);

(b) a second capture antibody which is specific to each of human pro B-type natriuretic peptide (“

human proBNP”

) and human brain natriuretic peptide (“

hBNP”

);

(c) a first detection antibody which is specific to each of human NT-proBNP and human proBNP; and

(d) a second detection antibody which is specific to hBNP.

10. The immunodiagnostic reagents of claim 9, wherein the first capture antibody is antibody 15F11.

11. The immunodiagnostic reagents of claim 9, wherein the second capture antibody is antibody 8.1.

12. The immunodiagnostic reagents of claim 9, wherein the first detection antibody is antibody 15C4.

13. The immunodiagnostic reagents of claim 9, wherein the second detection antibody is antibody 201.3.

14. A kit for detecting the amount of human NT-pro B-type natriuretic peptide

(“

human NT-proBNP”

), human pro B-type natriuretic peptide (“

human proBNP”

) and human brain natriuretic peptide (“

hBNP”

) in a test sample, said kit comprising;

(a) instructions for conducting the assay of the test sample; and

(b) immunodiagnostic reagents comprising;

(i) a first capture antibody specific for human NT-proBNP;

(ii) a second capture antibody which is specific to each of human proBNP and hBNP;

(iii) a first detection antibody which is specific to each of human NT-proBNP and human proBNP; and

(iv) a second detection antibody which is specific to hBNP.

15. The kit of claim 14, wherein the first capture antibody is antibody 15F11.

16. The kit of claim 14, wherein the second capture antibody is antibody 8.1.

17. The kit of claim 14, wherein the first detection antibody is antibody 15C4.

18. The kit of claim 14, wherein the second detection antibody is antibody 201.3.