ASSAYS FOR HUMAN NT-PRO B-TYPE NATRIURETIC PEPTIDE, HUMAN PRO B-TYPE NATRIURETIC PEPTIDE AND HUMAN B-TYPE NATRIURETIC PEPTIDE
First Claim
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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.
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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.
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Citations
18 Claims
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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. - View Dependent Claims (2, 3, 4)
- human NT-proBNP”
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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. - View Dependent Claims (6, 7, 8)
- human NT-proBNP”
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9. Immunodiagnostic reagents comprising:
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(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. - View Dependent Claims (10, 11, 12, 13)
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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. - View Dependent Claims (15, 16, 17, 18)
- human NT-proBNP”
Specification