Antibodies to troponin i and methods of use thereof

Abstract

The subject invention relates to antibodies to troponin I as well as methods of use thereof. In particular, such antibodies may be used to detect Troponin I in a patient and may also be used in the diagnosis of, for example, a myocardial infarction or acute coronary syndrome.

1 - 35 . (canceled) 
     
     
         36 . A composition comprising a detectable label bound to a Troponin I binding antibody, wherein said antibody comprises a CDR1 sequence of SEQ ID NO:52, a CDR2 sequence of SEQ ID NO: 53, and a CDR3 sequence of SEQ ID NO:54, and a light chain that comprises a CDR1 sequence of SEQ ID NO:55, a CDR2 sequence of SEQ ID NO: 56, and a CDR3 sequence of SEQ ID NO:57. 
     
     
         37 . A composition comprising a detectable label bound a Troponin I binding antibody, wherein said Troponin I binding antibody is a variant of TnI 19C7 and wherein variant comprises an antigen binding domain in which there is a mutation in the heavy chain CDR1 and CDR3 and a mutation in the light chain CDR1 and CDR2 as compared to the sequences of those respective CDRs in TnI 19C7 which consist of SEQ ID NO: 30 and SEQ ID NO:35 as CDR1 and CDR3 of TnI 19C7 heavy chain and SEQ ID NO: 40 and SEQ ID NO:45 as CDR1 and CDR2 of TnI 19C7 light chain, and wherein said antibody variant binds to Troponin I. 
     
     
         38 . The composition of  claim 36 , wherein said variant is a humanized antibody comprising a human acceptor framework. 
     
     
         39 . The composition of  claim 36 , wherein the amino acid sequence of the variable heavy chain of said antigen binding domain has at least 98% identity to SEQ ID NO:25. 
     
     
         40 . The composition of  claim 36 , wherein the amino acid sequence of the variable heavy chain of said antigen binding domain has at least 99% identity to SEQ ID NO:25. 
     
     
         41 . The composition of  claim 36 , wherein the amino acid sequence of the variable heavy chain of said antigen binding domain has a sequence of SEQ ID NO:25. 
     
     
         42 . The composition of  claim 36 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 94% identity to SEQ ID NO:28. 
     
     
         43 . The composition of  claim 36 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 95% identity to SEQ ID NO:28. 
     
     
         44 . The composition of  claim 36 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 96% identity to SEQ ID NO:28. 
     
     
         45 . The composition of  claim 36 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 97% identity to SEQ ID NO:28. 
     
     
         46 . The composition of  claim 36 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 98% identity to SEQ ID NO:28. 
     
     
         47 . The composition of  claim 36 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 99% identity to SEQ ID NO:28. 
     
     
         48 . The composition of  claim 36 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has a sequence of SEQ ID NO:28. 
     
     
         49 . The composition of  claim 36 , wherein said antibody variant comprises a heavy chain sequence has at least 98% identity to SEQ ID NO:25 and a light chain sequence that has at least 94% identity to SEQ ID NO:28. 
     
     
         50 . The composition of  claim 36 , wherein said detectable label is a signal-generating compound capable of generating a detectable signal selected from the group consisting of an enzyme, a prosthetic group complex, a fluorescent material, a luminescent material and a radioactive material. 
     
     
         51 . The composition of  claim 50 , wherein said enzymes are selected from the group consisting of horseradish peroxidase, alkaline phosphatase, beta-galactosidase, or acetylcholinesterase. 
     
     
         52 . The composition of  claim 50 , wherein said prosthetic group complex is selected from the group consisting of streptavidin/biotin and avidin/biotin. 
     
     
         53 . The composition of  claim 50 , wherein said fluorescent material is selected from the group consisting of umbelliferone, fluorescein, fluorescein isothiocyanate, rhodamine, dichlorotriazinylamine fluorescein, dansyl chloride and phycoerythrin. 
     
     
         54 . The composition of  claim 50 , wherein said luminescent material is luminol. 
     
     
         55 . The composition of  claim 50 , wherein said radioactive material is selected from the group consisting of 3H, 14C, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho, or 153Sm. 
     
     
         56 . The composition of  claim 50 , wherein said detectable label is acridinium. 
     
     
         57 . A composition comprising a solid support coated with a Troponin I binding antibody, wherein said antibody comprises a CDR1 sequence of SEQ ID NO:52, a CDR2 sequence of SEQ ID NO: 53, and a CDR3 sequence of SEQ ID NO:54, and a light chain that comprises a CDR1 sequence of SEQ ID NO:55, a CDR2 sequence of SEQ ID NO: 56, and a CDR3 sequence of SEQ ID NO:57. 
     
     
         58 . A composition comprising a solid support coated with a Troponin I binding antibody, wherein said Troponin I binding antibody is a variant of TnI 19C7 and wherein said variant comprises a mutation of at least one of heavy chain CDR1 of TnI 19C7 comprising SEQ ID NO: 30, heavy chain CDR3 of TnI 19C7 comprising SEQ ID NO:35, light chain CDR1 of TnI 19C7 comprising SEQ ID NO: 40, and light chain CDR2 of TnI 19C7 comprising of SEQ ID NO:45, and wherein said antibody variant binds to troponin I. 
     
     
         59 . The composition of  claim 58 , wherein said variant comprises a mutation in the heavy chain CDR1 and CDR3 and a mutation in the light chain CDR1 and CDR2 as compared to the sequences of those respective CDRs in TnI 19C7 which consist of SEQ ID NO: 30 and SEQ ID NO:35 as CDR1 and CDR3 of TnI 19C7 heavy chain and SEQ ID NO: 40 and SEQ ID NO:45 as CDR1 and CDR2 of TnI 19C7 light chain, and wherein said antibody variant binds to troponin I. 
     
     
         60 . The composition of  claim 57 , wherein said antibody variant comprises an antigen binding domain which comprises a heavy chain CDR1 sequence of SEQ ID NO: 52, a heavy chain CDR3 sequence of SEQ ID NO: 54 and a light chain CDR1 sequence of SEQ ID NO: 55 and a light chain CDR2 sequence of SEQ ID NO:56. 
     
     
         61 . The composition of  claim 57 , wherein said variant is a humanized antibody comprising a human acceptor framework. 
     
     
         62 . The composition of  claim 57 , wherein the amino acid sequence of the variable heavy chain of said antigen binding domain has at least 98% identity to SEQ ID NO:25. 
     
     
         63 . The composition of  claim 57 , wherein the amino acid sequence of the variable heavy chain of said antigen binding domain has at least 99% identity to SEQ ID NO:25. 
     
     
         64 . The composition of  claim 57 , wherein the amino acid sequence of the variable heavy chain of said antigen binding domain has a sequence of SEQ ID NO:25. 
     
     
         65 . The composition of  claim 57 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 94% identity to SEQ ID NO:28. 
     
     
         66 . The composition of  claim 57 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 95% identity to SEQ ID NO:28. 
     
     
         67 . The composition of  claim 57 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 96% identity to SEQ ID NO:28. 
     
     
         68 . The composition of  claim 57 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 97% identity to SEQ ID NO:28. 
     
     
         69 . The composition of  claim 57 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 98% identity to SEQ ID NO:28. 
     
     
         70 . The composition of  claim 57 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has at least 99% identity to SEQ ID NO:28. 
     
     
         71 . The composition of  claim 57 , wherein the amino acid sequence of the variable light chain of said antigen binding domain has a sequence of SEQ ID NO:28. 
     
     
         72 . The composition of  claim 57 , wherein said antibody variant comprises a heavy chain sequence has at least 98% identity to SEQ ID NO:25 and a light chain sequence that has at least 94% identity to SEQ ID NO:28. 
     
     
         73 . The composition of  claim 57 , wherein said solid support is selected from the group consisting of latex particles, magnetic particles, microparticles, beads, membranes, microtiter wells and plastic tubes. 
     
     
         74 . The composition of  claim 73 , wherein said solid support is magnetic. 
     
     
         75 . The composition of  claim 73 , wherein said solid support is a microparticle. 
     
     
         76 . A method of preparing a reagent for use in an immunoassay for detection of troponin I in a sample, comprising detectably labeling a Troponin I binding antibody, wherein said antibody comprises a CDR1 sequence of SEQ ID NO:52, a CDR2 sequence of SEQ ID NO: 53, and a CDR3 sequence of SEQ ID NO:54, and a light chain that comprises a CDR1 sequence of SEQ ID NO:55, a CDR2 sequence of SEQ ID NO: 56, and a CDR3 sequence of SEQ ID NO:57. 
     
     
         77 . A method of preparing a reagent for use in an immunoassay for detection of troponin I in a sample, comprising detectably labeling a Troponin I binding antibody, wherein said Troponin I binding antibody is a variant of TnI 19C7 and wherein variant comprises an antigen binding domain in which there is a mutation in the heavy chain CDR1 and CDR3 and a mutation in the light chain CDR1 and CDR2 as compared to the sequences of those respective CDRs in TnI 19C7 which consist of SEQ ID NO: 30 and SEQ ID NO:35 as CDR1 and CDR3 of TnI 19C7 heavy chain and SEQ ID NO: 40 and SEQ ID NO:45 as CDR1 and CDR2 of TnI 19C7 light chain, and wherein said antibody variant binds to troponin I. 
     
     
         78 . A method of preparing a reagent for use in an immunoassay for detection of troponin I in a sample, comprising conjugating to a solid support a Troponin I binding antibody, wherein said antibody comprises a CDR1 sequence of SEQ ID NO:52, a CDR2 sequence of SEQ ID NO: 53, and a CDR3 sequence of SEQ ID NO:54, and a light chain that comprises a CDR1 sequence of SEQ ID NO:55, a CDR2 sequence of SEQ ID NO: 56, and a CDR3 sequence of SEQ ID NO:57. 
     
     
         79 . A method of preparing a reagent for use in an immunoassay for detection of troponin I in a sample, comprising conjugating to a solid support a Troponin I binding antibody, wherein said Troponin I binding antibody is a variant of TnI 19C7 and wherein variant comprises an antigen binding domain in which there is a mutation in the heavy chain CDR1 and CDR3 and a mutation in the light chain CDR1 and CDR2 as compared to the sequences of those respective CDRs in TnI 19C7 which consist of SEQ ID NO: 30 and SEQ ID NO:35 as CDR1 and CDR3 of TnI 19C7 heavy chain and SEQ ID NO: 40 and SEQ ID NO:45 as CDR1 and CDR2 of TnI 19C7 light chain, and wherein said antibody variant binds to troponin I. 
     
     
         80 . A binding protein produced by culturing a host cell referred to as TnI 19C7 AM1 hG1 CHO 204, designated by American Type Culture Collection (ATCC) deposit Number PTA-9816 under conditions sufficient to produce said binding protein.