Thrombospondin Fragments and Uses Thereof In Clinical Assays for Cancer and Generation of Antibodies and Other Binding Agents

Abstract

The invention relates to thrombospondin fragments found in plasma, their use or use of portions thereof in diagnostic methods, as method calibrators, method indicators, and as immunogens, and as analytes for methods with substantial clinical utility; and their detection in plasma or other bodily fluids for purpose of diagnostic methods, especially for cancer.

1 - 66 . (canceled) 
     
     
         67 . A kit for the determination of the presence of, and/or the amount of, and/or the concentration of, one or more thrombospondin fragments in material taken or gathered from an individual, said kit comprising a binding agent capable of binding one or more of said fragments, said kit further comprising a reference molecule capable of binding to said binding agent, said reference molecule being selected from the group consisting of a thrombospondin fragment and a peptide derived from a thrombospondin fragment. 
     
     
         68 - 242 . (canceled) 
     
     
         243 . A kit of  claim 67  for the determination of the presence of, and/or the amount of, and/or the concentration of, a thrombospondin fragment or fragments in a material taken or gathered from an individual, said kit comprising a binding agent capable of binding a plasma thrombospondin fragment or plasma thrombospondin fragments, each of said plasma thrombospondin fragment or plasma thrombospondin fragments having a molecular weight not exceeding 140 kDa, wherein the size in kDa is that determined by gel electrophoresis after disulfide bond reduction, and wherein the plasma thrombospondin fragment or each of the plasma thrombospondin fragments comprises a region of thrombospondin, said region selected from the group of regions consisting of:
 a domain or a portion thereof within the protease-resistant core of thrombospondin, said domain being selected from the group consisting of a domain of inter-chain disulfide bonds, a procollagen-like domain, a type 1 repeat, a type 2 repeat, and a type 3 repeat; and 
 a collagen type V binding domain a or portion thereof, 
 and wherein the binding agent specifically binds to an epitope within said thrombospondin region, said kit further comprising a reference molecule, said reference molecule being selected from the group consisting of a thrombospondin fragment and a peptide derived from a thrombospondin fragment, said fragment or peptide corresponding to all or a portion of one or more regions selected from said group of regions, said reference molecule capable of binding to the binding agent. 
 
     
     
         244 . A kit of  claim 67  for the determination of the presence of, and/or the amount of, and/or the concentration of, a thrombospondin fragment or fragments in a material taken or gathered from an individual, said kit comprising a binding agent capable of binding an epitope, wherein the epitope is in a plasma thrombospondin fragment or portion thereof, said fragment or portion being at least 6 continuous amino acyl residues in length but of a molecular weight of 140 kDa or less, and wherein the reference molecule is selected from the group consisting of a thrombospondin fragment and a peptide derived from a thrombospondin fragment, said fragment or peptide being at least 6 continuous amino acyl residues in length but of a molecular weight of 140 kDa or less; wherein the size in kDa is that determined by gel electrophoresis after disulfide bond reduction. 
     
     
         245 . A kit of  claim 67  for the determination of the presence of, and/or the amount of, and/or the concentration of, a thrombospondin fragment or fragments in a material taken or gathered from an individual, said kit comprising a binding agent capable of binding an epitope, wherein the epitope is in a plasma thrombospondin fragment or fragments, said fragment or fragments being within a molecular weight range selected from the group consisting of 80 to 140 kDa, 40 to 60 kDa, and 20 to 35 kDa, and wherein the reference molecule is selected from the group consisting of a thrombospondin fragment and a peptide derived from a thrombospondin fragment, said fragment or peptide being within a molecular weight range selected from the group consisting of 80 to 140 kDa, 40 to 60 kDa, and 20 to 35 kDa, wherein the size in kDa is that determined by gel electrophoresis after disulfide bond reduction. 
     
     
         246 . A kit of  claim 67  for the determination of the presence of, and/or the amount of, and/or the concentration of, a thrombospondin fragment or fragments in a material taken or gathered from an individual, said kit comprising a binding agent capable of binding an epitope, wherein the epitope is in a plasma thrombospondin fragment or region thereof, provided that the epitope is not in a region selected from the group consisting of the fibrinogen-binding region in the amino-terminal domain of thrombospondin and a heparin-binding region sequence in the amino-terminal domain of thrombospondin; wherein the molecular weight of each of the fragment or fragments is at least 20 kDa but not more than 140 kDa, and wherein the reference molecule is selected from the group consisting of a thrombospondin fragment and a peptide derived from a thrombospondin fragment, and wherein the size in kDa is that determined by gel electrophoresis after disulfide bond reduction.