Diagnosis and monitoring of diseases

Abstract

The present invention relates to the diagnosis and monitoring of diseases and conditions by quantifying markers, including degradation products of disease-associated proteins, such as diketopiperazines composed of the two N-terminal amino acids or the two C-terminal amino acids of such proteins. The methods are useful for diagnosing or monitoring various diseases, including multiple sclerosis, Alzheimer's disease and ischemia. The invention further provides binding partners specific for the markers and compositions and kits for conducting the methods of the invention.

1 .- 46 . (canceled) 
     
     
         47 . A method for diagnosing or monitoring a disease or condition comprising the steps of:
 (a) obtaining a biological sample from a patient to be diagnosed or monitored;   (b) determining the quantity of a marker in said biological sample, wherein said marker is:
 (i) a truncated disease-associated protein lacking its two N-terminal amino acids, wherein said truncated disease-associated protein is not human serum albumin; 
 (ii) a truncated disease-associated protein lacking its two C-terminal amino acids; 
 (iii) a truncated disease-associated protein lacking its two N-terminal amino acids and its two C-terminal amino acids; 
 (iv) a diketopiperazine (DKP) comprising the two N-terminal amino acids of a disease-associated protein; 
 (v) a DKP comprising the two C-terminal amino acids of a disease-associated protein; or 
 (vi) two or more markers selected from those listed in (i) through (v) above; 
   
       provided that when only a single DKP is used as the marker, it will not be His-Pro DKP; and
 (c) determining if the quantity(ies) of said marker(s) in said biological sample is(are) indicative of the presence, absence or status of the disease or condition. 
 
     
     
         48 . The method of  claim 47 , wherein said disease-associated protein is myelin basic protein, beta-amyloid, Rh factor, pulmonary surfactant-associated protein A, B or D, insulin, tau protein, alpha-synuclein, albumin, C-reactive protein, interleukin 8, S100 proteins, beta-chorionic gonadotropin, fetal erythropoietin, pregnancy-associated protein A, myoglobin, troponin I, troponin T, prostate specific antigen, amylase, lipase, alphal-antitrypsyn, erthyropoietin, activated protein C, tethal chain, zeta chain, alpha chain, beta chain, delta chain, epsilon chain, gamma AG and brain natriuretic peptide. 
     
     
         49 . The method of  claim 47  or  48 , wherein said marker is a truncated disease-associated protein. 
     
     
         50 . The method of  claim 47  or  48 , wherein said marker is X-Y-DKP, wherein X-Y-DKP is a diketopiperazine composed of amino acids X and Y, and X and Y are the two N-terminal or the two C-terminal amino acids of a disease-associated protein. 
     
     
         51 . The method of  claim 50 , wherein X and Y are the two C-terminal amino acids of a disease-associated protein. 
     
     
         52 . The method of  claim 50 , wherein X and Y are the two N-terminal amino acids of a disease-associated protein. 
     
     
         53 . The method of  claim 50 , wherein X-Y-DKP is Asp-Ala-DKP, Met-Ala-DKP, Gln-Asn-DKP, Gly-Leu-DKP or combinations of the foregoing. 
     
     
         54 . The method of  claim 50 , wherein X-Y-DKP is Gly-Leu-DKP, Ala-Pro-DKP, Glu-Ala-DKP, Leu-Pro-DKP, Asp-Arg-DKP, His-Gly-DKP or combinations of the foregoing. 
     
     
         55 . The method of  claim 50 , wherein X-Y-DKP is Arg-His-DKP, His-Pro-DKP, Ser-Pro-DKP, or combinations of the foregoing. 
     
     
         56 . The method of  claim 50 , wherein X-Y-DKP is Gly-Leu-DKP, Pro-Glu-DKP, Gln-Gly-DKP, Glu-Ser-DKP, or combinations of the foregoing. 
     
     
         57 . The method of  claim 47 , wherein the disease or condition is multiple sclerosis, rheumatoid arthritis, acute respiratory distress syndrome, cystic fibrosis, diabetes mellitus, Alzheimer's disease, Parkinson's disease, inflammation, ischemia, cerebral ischemia, placental ischemia, myocardial infarction, prostate cancer, pancreatitis, emphysema, renal disease, cancer, chemotherapy, hemoglobinopathies, anemnias or congestive heart failure. 
     
     
         58 . The method of  claim 57  wherein the disease or condition is Alzheimer's disease. 
     
     
         59 . The method of  claim 58 , wherein the method comprises determining the quantity in said biological sample of Asp-Ala-DKP. 
     
     
         60 . The method of  claim 59  further comprising determining the quantity of a compound having a mass of about 175 as determined by liquid chromatography and mass spectrometry. 
     
     
         61 . The method of  claim 57  wherein the disease or condition is placental ischemia. 
     
     
         62 . The method of  claim 61 , wherein the method comprises determining the quantity in said biological sample of:
 (i) Gly-Leu-DKP;   (ii) Ala-Pro-DKP; or   (iii) both Gly-Leu-DKP and Ala-Pro-DKP.   
     
     
         63 . The method of  claim 47 , wherein step (b) is conducted by mass spectrometry, chemical assay or immunoassay. 
     
     
         64 . The method of  claim 63 , wherein step (b) is conducted by immunoassay. 
     
     
         65 . The method of  claim 64 , wherein said immunoassay is conducted by using one or more binding partners specific for a marker. 
     
     
         66 . The method of  claim 65 , wherein the binding partner is an antibody or an aptamer. 
     
     
         67 . The method of  claim 47 , wherein said biological sample is a body fluid. 
     
     
         68 . The method of  claim 67 , wherein said body fluid is serum, plasma, blood, urine, saliva, cerebrospinal fluid, tears, semen, vaginal secretion, amniotic fluid or cord blood. 
     
     
         69 . The method of  claim 68 , wherein said body fluid is plasma or serum. 
     
     
         70 . The method of  claim 47 , wherein said patient is an animal. 
     
     
         71 . The method of  claims 70 , wherein said patient is a human. 
     
     
         72 . An isolated binding partner having specificity for a marker selected from the group consisting of:
 (a) a truncated disease-associated protein lacking its two N-terminal amino acids, wherein said truncated disease-associated protein is not human serum albumin;   (b) a truncated disease-associated protein lacking its two C-terminal amino acids;   (c) a truncated disease-associated protein lacking its two N-terminal amino acids and its two C-terminal amino acids;   (d) a diketopiperazine (DKP) comprising the two N-terminal amino acids of a disease-associated protein, wherein the DKP is not His-Pro DKP; and   (e) a DKP comprising the two C-terminal amino acids of a disease-associated protein, wherein the DKP is not His-Pro DKP.   
     
     
         73 . The isolated binding partner of  claim 72 , wherein the binding partner has specificity for a DKP. 
     
     
         74 . The isolated binding partner of  claim 73 , wherein the binding partner has specificity for Arg-Arg-DKP, Gln-Asn-DKP, Lys-Arg-DKP, Glu-Phe-DKP, Ser-Met-DKP, Cys-Asn-DKP, Lys-Ala-DKP, Gln-Asn-DKP, Gly-Leu-DKP, Ala-Ala-DKP, Trp-Pro-DKP, Asn-Ser-DKP, Leu-Pro-DKP, Asp-Arg-DKP, His-Gly-DKP, Gln-Gly-DKP, Glu-Ser-DKP, Asn-Pro-DKP, Lys-Leu-DKP, Pro-Cys-DKP, Asn-Lys-DKP, Asp-Arg-DKP, Ala-Pro-DKP or Arg-His-DKP. 
     
     
         75 . The isolated binding partner of  claim 73 , wherein the binding partner has specificity for N-acetyl-Ala-Ser-DKP, N-acetyl-Ala-phosphorylated-Ser-DKP, Asp-Ala-DKP, Glu-Ile-DKP, Glu-Val-DKP, Phe-Pro-DKP, Ala-Glu-DKP, Phe-Val-DKP, Gly-Ile-DKP, Met-Ala-DKP, Met-Asp-DKP, Glu-Lys-DKP, Gln-Thr-DKP, Ala-Val-DKP, Gly-Leu-DKP, Ala-Pro-DKP, Glu-Ala-DKP, Pro-Glu-DKP, Lys-Ser-DKP, Ile-Val-DKP, Gln-Tyr-DKP, Lys-Glu-DKP, Glu-Asp-DKP, Ala-Pro-DKP, Ala-Asn-DKP, Ala-Leu-DKP, Ser-Leu-DKP, Val-Leu-DKP, Val-His-DKP, Gly-His-DKP or Ser-Pro-DKP. 
     
     
         76 . The isolated binding partner of any one of  claims 72 - 75 , wherein said binding partner is an antibody. 
     
     
         77 . The isolated binding partner of  claim 76 , wherein said antibody is a monoclonal antibody. 
     
     
         78 . The isolated binding partner of any one of  claims 72 - 75 , wherein said binding partner is an aptamer. 
     
     
         79 . A composition comprising the binding partner of any one of  claims 72 - 75  in a physiologically-acceptable carrier. 
     
     
         80 . A kit comprising the binding partner of  claim 72 - 75  and associated reagents for quantitating the marker. 
     
     
         81 . The kit of  claim 80 , wherein said binding partner is an antibody. 
     
     
         82 . The kit of  claim 81 , wherein said antibody is a monoclonal antibody. 
     
     
         83 . The kit of  claim 80 , wherein said binding partner is an aptamer.