Assays for natriuretic peptide NT-pro-human type b, the pro-natriuretic peptide and human type b-type natriuretic peptide human b

摘要

An immunoassay for quantifying the amount of natriuretic NT-pro-peptide of human type B ( "NT-proBNPhumano"), pro-peptide ( "proBNP human") natriuretic human type B and natriuretic peptide cerebral5 human ( "hBNP" ) in a test sample being tested or suspected of containing the NT-proBNPhumano, human proBNP and hBNP, the method comprising the steps of: (a) contacting a test sample with (i) a first capture antibody that binds to the NT-proBNPhumano and that has been immobilized on a solid phase to produce a first immobilized antibody yformar a first mixture comprising a complex of first capture antibody-human NT-proBNP; (Ii) a second capture antibody that binds human proBNP and that has been immobilized on a fasesólida to produce a second immobilized antibody and form a second mixture comprising uncomplejo the second capture antibody-human proBNP; and (iii) the second capture antibody, queademás to bind human proBNP also binds to hBNP which has been immobilized on a phase sólidapara produce a third immobilized antibody and form a third mixture comprising a delsegundo complex capture antibody-hBNP human, wherein said first capture antibody comprises elanticuerpo 15F11 and second capture antibody comprising antibody 8.1; (B) contacting said first mixture comprising the complex of first capture antibody-human proBNP with a first detection antibody which binds human proBNP and has sidoconjugado with a detectable label to form a third mixture comprising a complex of capture-primeranticuerpo NTproBNP first human-detection antibody, wherein the first antibody is the 15C4 antibody dedetección; (C) contacting said depending on the mixture comprising the complex of the second antibody capturaproBNPhumano with the first antibody detection, in addition to binding to NT-proBNP human, tambiénse binds human proBNP, and that has been conjugated to a label detectable to form a fifth mezclaque comprises a second antibody complex human-first-proBNP capture detection antibody, wherein the first antibody detection is the antibody 15C4; (D) contacting the third mixture comprising the complex of the second antibody capture-hBNPhumano with a second detecting antibody that binds to hBNP and that has been conjugated to a marcadordetectable to form a sixth mixture comprising a second complex capture antibody-detection hBNPsegundoanticuerpo, wherein the second detection antibody is the antibody 201.3; and (e) determining the amount of (i) the complex of first capture antibody-NT-proBNP human-primeranticuerpo detection formed in step (b) by detecting the detectable label as unamedida the amount of NT-proBNP human contained in the test sample; (Ii) the second complex deanticuerpo human proBNP capture-detection-first antibody complex formed in step (c) by the detectable label ladetección as a measure of the amount of human proBNP contained in the test muestrade; and (iii) the complex of second capture antibody-hBNP-second antibody detection formadoen step (d) by detecting the detectable label as a measure of the amount of hBNPcontenido in the test sample.