Detection and determination of the stages of coronary artery disease

Abstract

A method having clinically sufficient degree of diagnostic accuracy for detecting the presence of coronary artery disease in a human patient from the general population and for distinguishing between the stages of the disease in that patient is disclosed. The stages are, first, the non-acute stage, which is either asymptomatic coronary artery disease or stable angina, second, the acute stage known as unstable angina, and, third, the acute stage known as acute myocardial infarction. The diseased state (as opposed to the non-diseased state) is indicated by the clinically significant presence of a first marker in a sample from the patient. The presence of one of the two acute stages, unstable angina or acute myocardial infarction, is indicated by the clinically significant presence of a second marker in a sample from the patient. The presence of the more severe acute stage known as acute myocardial infarction is indicated by the clinically significant presence of a third marker in a sample from the patient. Preferably the first marker comprises OxLDL, the second marker comprises MDA-modified LDL, and the third marker is a troponin. Preferably the OxLDL and MDA-modified LDL are detected using monoclonal antibodies that can detect the presence of those markers in undiluted human plasma at concentrations as low as 0.02 milligrams/deciliter.

We claim:  
     
         1 . A method having a clinically sufficient degree of diagnostic accuracy for detecting the presence of and for distinguishing between or among the non-acute and the acute stages of coronary artery disease for a human patient from the general population, the non-acute stage of coronary artery disease being either asymptomatic coronary artery disease or stable angina and the acute stages of coronary artery disease being unstable angina and acute myocardial infarction, the method comprising performing step (b) and performing at least one of steps (a) and (c): 
 (a) testing a sample from the patient for a clinically significant presence of a first marker whose presence above a predetermined level can indicate with a very high degree of diagnostic accuracy the presence of coronary artery disease;    (b) testing a sample from the patient for a clinically significant presence of second marker whose presence above a predetermined level can indicate with a very high degree of diagnostic accuracy the presence of an acute stage of coronary artery disease; and    (c) testing a sample from the patient for a clinically significant presence of a third marker whose presence above a predetermined level can indicate with a high degree of diagnostic accuracy the presence of acute myocardial infarction.    
     
     
         2 . The method of    claim 1    wherein the first marker is a first atherogenic protein.  
     
     
         3 . The method of    claim 2    wherein the first atherogenic protein comprises OxLDL containing at least 60 substituted lysine residues per apo B-100 moiety.  
     
     
         4 . The method of claim I wherein the second marker is a second atherogenic protein.  
     
     
         5 . The method of    claim 4    wherein the second atherogenic protein comprises MDA-modified LDL containing at least 60 substituted lysine residues per apo B-100 moiety.  
     
     
         6 . The method of    claim 1    wherein the third marker is a heart protein.  
     
     
         7 . The method of    claim 6    wherein the third marker is selected from the group consisting of a troponin and CK-MB.  
     
     
         8 . The method of    claim 1    wherein step (a) uses an immunological assay that can indicate the clinically significant presence of the first marker.  
     
     
         9 . The method of    claim 8    wherein the immunological assay is a first sandwich assay.  
     
     
         10 . The method of    claim 8    wherein the immunological assay uses a first monoclonal antibody having a high affinity for the first marker.  
     
     
         11 . The method of    claim 10    wherein the first monoclonal antibody has an affinity for the first marker of at least about 1×10 10  M −1 .  
     
     
         12 . The method of    claim 11    wherein the first monoclonal antibody is mAb-4E6.  
     
     
         13 . The method of    claim 1    wherein step (b) is conducted using an immunological assay that can indicate the clinically significant presence of the second marker.  
     
     
         14 . The method of    claim 13    wherein the immunological assay is a second sandwich assay.  
     
     
         15 . The method of    claim 13    wherein the immunological assay uses a second monoclonal antibody having a high affinity for the second marker.  
     
     
         16 . The method of    claim 15    wherein the second monoclonal antibody has an affinity for the second marker of at least about 1×10 10  M −1 .  
     
     
         17 . The method of    claim 16    wherein the second monoclonal antibody is mAb-1H11.  
     
     
         18 . The method of    claim 1    wherein step (a) is performed.  
     
     
         19 . The method of    claim 18    wherein the first marker is a first atherogenic protein comprising OxLDL, and the second marker is a second atherogenic protein comprising MDA-modified LDL.  
     
     
         20 . The method of    claim 18    wherein steps (a) and (b) are performed using immunological assays.  
     
     
         21 . The method of    claim 20    wherein the assays are performed using monoclonal antibodies selected from the group consisting of mAb-4E6, mAb-1H11, and mAb-8A2.  
     
     
         22 . The method of    claim 20    wherein the immunological assays are sandwich assays.  
     
     
         23 . The method of    claim 22    wherein the assays are performed using monoclonal antibodies selected from the group consisting of mAb-4E6, mAb-1H11, and mAb-8A2.  
     
     
         24 . The method of    claim 19    wherein the test of step (a) can detect the presence of OxLDL containing at least 60 substituted lysine residues per apo B-100 moiety in undiluted human plasma in a concentration of 0.02 milligrams/deciliter and the test of step (b) can detected the presence of MDA-modified LDL containing at least 60 substituted lysine residues per apo B-100 moiety in undiluted human plasma in a concentration of 0.02 milligrams/deciliter.  
     
     
         25 . The method of    claim 1    wherein step (c) is performed.  
     
     
         26 . The method of    claim 25    wherein the third marker is a heart protein.  
     
     
         27 . The method of    claim 25    wherein the third marker is selected from the group consisting of a troponin and CK-MB.  
     
     
         28 . The method of    claim 1    wherein both of steps (a) and (c) are performed.  
     
     
         29 . The method of    claim 28    wherein the first marker is a first atherogenic protein.  
     
     
         30 . The method of    claim 29    wherein the first atherogenic protein comprises OxLDL containing at least 60 substituted lysine residues per apo B-100 moiety.  
     
     
         31 . The method of    claim 28    wherein the second marker is a second atherogenic protein.  
     
     
         32 . The method of    claim 31    wherein the second atherogenic protein comprises MDA-modified LDL containing at least 60 substituted lysine residues per apo B-100 moiety.  
     
     
         33 . The method of    claim 28    wherein the third marker is a heart protein.  
     
     
         34 . The method of    claim 33    wherein the third marker is selected from the group consisting of a troponin and CK-MB.  
     
     
         35 . The method of    claim 28    wherein step (a) uses an immunological assay that can indicate the clinically significant presence of the first marker.  
     
     
         36 . The method of    claim 35    wherein the immunological assay is a first sandwich assay.  
     
     
         37 . The method of    claim 35    wherein the immunological assay uses a first monoclonal antibody having a high affinity for the first marker.  
     
     
         38 . The method of    claim 37    wherein the first monoclonal antibody has an affinity for the first marker of at least about 1×10 10  M −1 .  
     
     
         39 . The method of    claim 38    wherein the first monoclonal antibody is mAb-4E6.  
     
     
         40 . The method of    claim 28    wherein step (b) is conducted using an immunological assay that can indicate the clinically significant presence of the second marker.  
     
     
         41 . The method of    claim 40    wherein the immunological assay is a second sandwich assay.  
     
     
         42 . The method of    claim 40    wherein the immunological assay uses a second monoclonal antibody having a high affinity for the second marker.  
     
     
         43 . The method of    claim 42    wherein the second monoclonal antibody has an affinity for the second marker of at least about 1×10 10  M −1 .  
     
     
         44 . The method of    claim 43    wherein the second monoclonal antibody is mAb-1H11.  
     
     
         45 . The method of    claim 28    wherein the first marker is a first atherogenic protein comprising OxLDL containing at least 60 substituted lysine residues per apo B-100 moiety and the second marker is a second atherogenic protein comprising MDA-modified LDL containing at least 60 substituted lysine residues per apo B-100 moiety.  
     
     
         46 . The method of    claim 45    wherein steps (a) and (b) are conducted using immunological assays.  
     
     
         47 . The method of    claim 46    wherein the immunological assays are sandwich assays.  
     
     
         48 . The method of    claim 46    wherein the assays are conducted using monoclonal antibodies selected from the group consisting of mAb-4E6, mAb-1H11, and mAb-8A2.  
     
     
         49 . The method of    claim 45    wherein the test of step (a) can detect the presence of OxLDL containing at least 60 substituted lysine residues per apo B-100 moiety in undiluted human plasma in a concentration of 0.02 milligrams/deciliter and the test of step (b) can detected the presence of MDA-modified LDL containing at least 60 substituted lysine residues per apo B-100 moiety in undiluted human plasma in a concentration of 0.02 milligrams/deciliter.  
     
     
         50 . The method of    claim 49    wherein the third marker is selected from the group consisting of a troponin and CK-MB.  
     
     
         51 . A method having a clinically sufficient degree of diagnostic accuracy for detecting the presence of and for distinguishing between or among the non-acute and the acute stages of coronary artery disease for a human patient from the general population, the non-acute stage of coronary artery disease being either asymptomatic coronary artery disease or stable angina and the acute stages of coronary artery disease being unstable angina and acute myocardial infarction, the method comprising the steps: 
 (a) testing a sample from the patient using an immunological assay for a clinically significant presence of OxLDL containing at least 60 substituted lysine residues per apo B-100 moiety, its presence above a predetermined level being able to indicate with a very high degree of diagnostic accuracy the presence of coronary artery disease, the assay employing at least one monoclonal antibody having a high affinity for the OxLDL;    (b) testing a sample from the patient using an immunological assay for a clinically significant presence of MDA-modified LDL containing at least 60 substituted lysine residues per apo B-100 moiety, its presence above a predetermined level being able to indicate with a very high degree of diagnostic accuracy the presence of an acute stage of coronary artery disease, the assay employing at least one monoclonal antibody having a high affinity for MDA-modified LDL; and    (c) optionally testing a sample from the patient for a clinically significant presence of a third marker whose presence above a predetermined level can indicate with a high degree of diagnostic accuracy the presence of acute myocardial infarction.    
     
     
         52 . The method of    claim 51    wherein the assay of step (a) can detect the presence of the OxLDL in undiluted human plasma in a concentration of 0.02 milligrams/deciliter and the assay of step (b) can detected the presence of the MDA-modified LDL in undiluted human plasma in a concentration of 0.02 milligrams/deciliter.  
     
     
         53 . The method of    claim 51    wherein the at least one monoclonal antibody used in the assay of step (a) has an affinity for the OxLDL of at least about 1×10 10  M −1  and the at least one monoclonal antibody used in the assay of step (b) has an affinity for the MDA-modified LDL of at least about 1×10 10  M −1 .  
     
     
         54 . The method of    claim 51    wherein the monoclonal antibodies used in the assays of steps (a) and (b) are selected from the group consisting of mAb-4E6, mAb-1H11, and mAb-8A2.  
     
     
         55 . The method of    claim 51    wherein step (c) is performed and the third marker is a heart protein.  
     
     
         56 . The method of    claim 55    wherein the heart protein is selected from the group consisting of a troponin and CK-MB.  
     
     
         57 . A method having a clinically sufficient degree of diagnostic accuracy for detecting the presence of and for distinguishing between or among the non-acute and the acute stages of coronary artery disease for a human patient from the general population, the non-acute stage of coronary artery disease being either asymptomatic coronary artery disease or stable angina and the acute stages of coronary artery disease being unstable angina and acute myocardial infarction, the method comprising the steps: 
 (a) testing a sample from the patient using an immunological assay for a clinically significant presence of OxLDL containing at least 60 substituted lysine residues per apo B-100 moiety, its presence above a predetermined level being able to indicate with a very high degree of diagnostic accuracy the presence of coronary artery disease, the assay employing at least one monoclonal antibody having a high affinity for the OxLDL;    (b) testing a sample from the patient using an immunological assay for a clinically significant presence of MDA-modified LDL containing at least 60 substituted lysine residues per apo B-100 moiety, its presence above a predetermined level being able to indicate with a very high degree of diagnostic accuracy the presence of an acute stage of coronary artery disease, the assay employing at least one monoclonal antibody having a high affinity for MDA-modified LDL; and    (c) testing a sample from the patient for a clinically significant presence of a heart protein whose presence above a predetermined level can indicate with a high degree of diagnostic accuracy the presence of acute myocardial infarction.    
     
     
         58 . The method of    claim 57    wherein the monoclonal antibodies used in the assays of steps (a) and (b) are selected from the group consisting of mAb-4E6, mAb-1H11, and mAb-8A2 and the heart protein is selected from the group consisting of a troponin and CK-MB.