Methods for the assay of troponin I and T and selection of antibodies for use in immunoassays
DCFirst Claim
1. An assay for determining the presence or amount of a cardiac specific free and complexed isoform of troponin, said complex comprising isoforms selected from the group consisting of troponin I, troponin C and troponin T, comprising the steps of performing an immunoassay with antibody specific to a cardiac troponin which binds a complex comprising two or more of said isoform and another of said isoform of troponin in said sample, wherein the results of said immunoassay indicate the total amount of said free and complexed cardiac specific troponin isoform in said blood sample.
12 Assignments
Litigations
0 Petitions
Accused Products
Abstract
Assays and antibodies are disclosed for the detection and quantitation of cardiac specific troponin I and troponin T in body fluids as an indicator of myocardial infarction. Since troponin I and T exist in various conformations in the blood, the ratios of the monomeric troponin I and T and the binary and ternary complexes may be related to the metabolic state of the heart. More specifically, immunoassays for determining the presence or amount of free, binary and ternary complexes of troponin I and T are claimed.
242 Citations
18 Claims
- 1. An assay for determining the presence or amount of a cardiac specific free and complexed isoform of troponin, said complex comprising isoforms selected from the group consisting of troponin I, troponin C and troponin T, comprising the steps of performing an immunoassay with antibody specific to a cardiac troponin which binds a complex comprising two or more of said isoform and another of said isoform of troponin in said sample, wherein the results of said immunoassay indicate the total amount of said free and complexed cardiac specific troponin isoform in said blood sample.
-
6. An immunoassay method for determining the presence or amount of complexes of a cardiac specific isoform of troponin in a blood sample, said method comprising performing step a) or b), and steps c) through e) as follows:
-
a) providing an antibody conjugate comprising an antibody which specifically binds to the cardiac specific isoform of troponin in a complex comprising said isoform and another isoform of troponin, and a means for generating a signal;
or,b) providing an antibody conjugate comprising an antibody which specifically binds to the cardiac specific isoform of troponin in a binary complex comprising said isoform and another isoform of troponin, and a means for generating a signal; and
, providing an antibody conjugate comprising an antibody which specifically binds to a cardiac specific isoform of troponin I or T, or troponin C in a ternary complex of three isoforms of troponin, and a means for generating a signal; and
,c) contacting said sample with said antibody conjugate of step a), or with the antibody conjugates of step b), whereby the antibody conjugates bind to the troponin complexes present in the sample; d) contacting the sample with a solid phase to which is bound at least one capture antibody which specifically binds said troponin complexes which are bound to antibody conjugate or bound to troponin complexes which is bound to antibody conjugate following binding to the capture antibody, whereby a detectable signal is present from the antibody conjugate upon binding to the solid phase of cardiac specific troponin complexes which are or become bound to antibody conjugate; and
,e) relating the detectable signal to the total amount of the cardiac specific isoform of troponin in said sample.
-
-
7. An immunoassay method for determining the presence or amount of complexes comprising two or more isoforms of troponin, of any cardiac specific isoform of troponin in a blood sample, said method comprising performing step a) or b), and steps c) through f) as follows:
-
a) providing an antibody conjugate comprising an antibody which specifically binds to troponin complexes comprising two or more troponin isoforms, and a means for generating a signal;
or,b) providing an antibody conjugate comprising an antibody which specifically binds to troponin in binary complexes comprising two troponin isoforms, and a means for generating a signal;
or, providing an antibody conjugate comprising an antibody which specifically binds to ternary complexes comprising three troponin isoforms, and a means for generating a signal; and
,c) contacting said sample with said antibody conjugate of step a), or with the antibody conjugate(s) of step b), whereby the antibody conjugates bind to troponin complexes present in the sample; d) contacting the sample with a solid phase to which is bound at least one capture antibody which specifically binds a cardiac specific isoform of troponin in complexes which are bound to antibody conjugate, or, with a solid phase to which is bound at least one capture antibody which specifically binds to a cardiac specific isoform of troponin which is bound to antibody conjugate after binding to capture antibody, whereby a detectable signal is present from the antibody conjugate upon binding to the solid phase of the cardiac specific troponin isoform which is or becomes bound to antibody conjugate; and
,e) relating the detectable signal to the presence or amount of the cardiac specific isoform of troponin in said sample.
-
-
8. An assay for determining the presence or amount of all free cardiac specific troponin isoforms and complexed isoforms of cardiac troponin in a blood sample, said complexes comprising isoforms selected from the group consisting of troponin I, troponin C and troponin T, comprising the steps of:
- performing an immunoassay with antibody specific to cardiac troponin which binds a complex comprising two or more of said isoforms of troponin in said sample, wherein the results of said immunoassay indicate the total amount of said free and complexed cardiac specific troponin isoforms in said blood sample.
- View Dependent Claims (10)
-
9. An assay for determining the presence or amount of complexed isoforms of cardiac troponin in a blood sample, said complexes comprising isoforms selected from the group consisting of troponin I, troponin C and troponin T, comprising the steps of:
- performing an immunoassay with antibody specific to cardiac troponin which binds a complex comprising two or more of said isoforms of troponin in said sample wherein the results of said immunoassay indicate the total amount of said complexed cardiac specific isoforms in said blood sample.
-
15. An assay for determining the presence or amount of a free and complexed cardiac specific isoform of troponin in a blood sample, said complex comprising isoforms selected from the group consisting of troponin I, troponin C and troponin T, comprising the steps of performing an immunoassay with antibody specific to cardiac troponin which binds said isoform and a complex comprising two or more of said isoform and another of said isoform of troponin in said sample, wherein the results of said immunoassay indicate the total amount of said free and complexed cardiac specific troponin isoform in said blood sample.
-
16. An assay for determining the presence or amount of a free and complexed cardiac specific isoform of troponin in a blood sample, said complex comprising isoforms selected from the group consisting of troponin I, troponin C and troponin T, comprising the steps of performing an immunoassay with antibody specific to cardiac troponin I and/or T which binds a complex comprising two or more of said isoform and another of said isoform of troponin in said sample, wherein the results of said immunoassay indicate the total amount of said free and complexed cardiac specific troponin isoform in said blood sample.
-
17. An assay for determining the presence or amount of a free and complexed cardiac specific troponin I in a blood sample, said complex comprising isoforms selected from the group consisting of troponin I, troponin C and troponin T, comprising the steps of performing an immunoassay with antibody specific to cardiac troponin I which binds a complex comprising two or more isoforms, one of which is troponin I and another of said isoform of troponin in said sample, wherein the results of said immunoassay indicate the total amount of said free and complexed cardiac specific troponin I isoform in said blood sample.
-
18. An assay for determining the presence or amount of a free and complexed cardiac specific troponin T in a blood sample, said complex comprising isoforms selected from the group consisting of troponin I, troponin C and troponin T, comprising the steps of performing an immunoassay with antibody specific to cardiac troponin T which binds a complex comprising two or more isoforms, one of which is troponin T and another of said isoform of troponin in said sample, wherein the results of said immunoassay indicate the total amount of said free and complexed cardiac specific troponin T isoform in said blood sample.
Specification