Abstract: A method of bioassay for the quantification of biomarkers useful in the diagnosis of fibrosis disease and prognosis of its development, including biomarkers indicative of the risk of developing fibrosis after a chronic injury is provided. In particular, according to the present invention, biomarkers relating to degradation fragments of Collagen type I, III, IV, V, and VI, elastin, C-reactive protein, and proteoglycans including Biglycan, Decorin, Versican, and Perlecan are found to be useful.

Abstract: Provided herein is a sandwich immunoassay for detecting cross-linked PIIINP that has at least two strands of PIIINP joined together by inter-strand cross-linking each having a C-terminal neo-epitope of PIIINP that is generated by N-protease cleavage of intact type III procollagen. A biological sample having the cross-linked PIIINP is contacted with a first surface-bound monoclonal antibody and then by a second monoclonal antibody, both specifically reactive with a neoepitope in the C-terminal sequence of PIIINP, and then binding of the second monoclonal antibody is determined. Also provided is a method for evaluating the efficacy of an antagonist drug targeting lysyl oxidases via the immunoassay and a kit containing a solid support binding the first monoclonal antibody and containing the second monoclonal antibody.

Abstract: Provided herein is a method of bioassay for the quantification of peptide fragments relevant to neurodegenerative conditions. The method comprises cleaving a Tau protein by a secretase, such as ADAM10, to form a neo-epitope. The method comprises contacting a blood derived sample with an antibody specific for the neo-epitope and determining the level of binding of the antibody to peptide fragments comprising the neo-epitope in the sample. Neo-epitope containing peptide levels are found to be inversely correlated to cognitive function.

Abstract: Provided is a monoclonal antibody specifically reactive with a C-terminal neo-epitope of PIIINP comprised in a C-terminal amino acid sequence CPTGXQNYSP-COOH (SEQ ID NO: 4) in which X is Gly or Pro, and where the monoclonal antibody does not recognize or bind an elongated version of the C-terminal amino acid sequence CPTGXQNYSPQZ-COOH (SEQ ID NO: 5), in which Z is absent or is one or more amino acids of the sequence of collagen type III. Also provided is a method of immunoassay for detecting in a biological sample the C-terminal neo-epitope of PIIINP generated by N-protease cleavage of intact type III procollagen, by contacting the sample with the monoclonal antibody, and determining the amount of binding of the antibody.

Abstract: Methods of diagnosis or of quantitation of fibrosis are provided that comprise conducting an immunoassay to measure neo-epitope containing protein fragments naturally present in a bioflui sample, and associating an elevation of the measure in the patient above a normal level with the presence or extent of fibrosis. The immunoassay is conducted by a contacting protein fragments naturally present in the sample with an immunological binding partner reactive with a neo-epitope formed by cleavage of a protein by a proteinase and measuring the extent of binding of peptide fragments to the immunological binding partner to measure therein protein fragments comprising the neo-epitope. The protein is collagen type I, collagen type IV, collagen type V, elastin, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, vimentin, or C-reactive protein.

Abstract: Provided herein are methods of diagnosis or of quantitation of fibrosis. An immunoassay is conducted to measure neo-epitope containing protein fragments of collagen type III, collagen type I, collagen type IV, collagen type V, or collagen type VI, elastin, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, vimentin, or C-reactive protein naturally present in a biofluid sample obtained from a patient. An above normal elevation of the measured protein fragments in the patient is associated with the presence or extent of fibrosis.

Abstract: An immunoassay for the quantification of fragments having an N- or a C-terminal neo-epitope formed by cleavage of a titin protein by a proteinase is provided. The immunoassay includes the steps of contacting a sample with an antibody specifically binding to the N- or C-terminal neo-epitope of the fragments and determining the level of binding.

Abstract: Provided is a method of bioassay for the quantification of peptide fragments elevated in lung diseases such as COPD, SCC, or IPF. The peptide fragments comprise a neo-epitope formed at a cleavage site by cleavage in vivo of elastin by a proteinase. In the method a sample is contacted with an antibody having specific binding affinity for the neo-epitope amino acid sequence and determining the level of binding is determined where the antibody binds one of the following terminal sequences: . . . FGPGVV, . . . VPGLGV or IKAPKL . . . . Also provided are antibodies and immunoassay kits for use in such methods.

Abstract: An assay for citrullinated fragments of SOCS-2, Alpha 1 anti tyrpsin, versican, biglycan, laminin, or other protein having a terminal antibody binding site comprising citrulline in a blood derived sample shows diagnostic relevance in relation to rheumatoid arthritis or fibrotic disease.

Abstract: Provided is a method of bioassay for the quantification of peptide fragments comprising a neo-epitope formed by cleavage of a protein of an atherosclerotic plaque such as lumican, versican, perlecan, deocrin, biglycan, collagen type III, CRP, ApoE, or elastin by a proteinase. The method comprises contacting a sample such as urine or serum with an antibody reactive with the neo-epitope and determining the level of binding of the immunological binding partner to peptide fragments in the sample. The assay is predictive of risk of cardiovascular disease events.

Abstract: Provided is a method for measuring the propensity of a patient to suffer progression of Alzheimer's disease that comprises, in either order: a) conducting a first immunoassay of a patient sample to determine the content (Tau-C?) of a C-terminal epitope present at the terminus of the amino acid sequence -SSTGSIDMVD (SEQ ID NO:1), and b) conducting a second immunoassay of the sample to determine the content (Tau-A?) of an N-terminal epitope present at the terminus of the amino acid sequence TPRGAAPPGQ- (SEQ ID NO:2), and c) calculating an index as a ratio Tau-A?/Tau-C? between the second level and the first level. The method may be used to select patients for a clinical trial of an intervention in Alzheimer's disease or to evaluate a therapy.

Abstract: Provided herein is a sandwich immunoassay for detecting cross-linked PIIINP that has at least two strands of PIIINP joined together by inter-strand cross-linking each having a C-terminal neo-epitope of PIIINP that is generated by N-protease cleavage of intact type III procollagen. A biological sample having the cross-linked PIIINP is contacted with a first surface-bound monoclonal antibody and then by a second monoclonal antibody, both specifically reactive with a neoepitope in the C-terminal sequence of PIIINP, and then binding of the second monoclonal antibody is determined. Also provided is a method for evaluating the efficacy of an antagonist drug targeting lysyl oxidases via the immunoassay and a kit containing a solid support binding the first monoclonal antibody and containing the second monoclonal antibody.

Abstract: Methods of diagnosis or of quantitation of pathological conditions comprise conducting an immunoassay to measure neo-epitope containing protein fragments naturally present in a biofluid sample, and associating an elevation of the measure in the patient above a normal level with the presence or extent of pathology. The immunoassay is conducted by a method comprising: contacting protein fragments naturally present in the sample with an immunological binding partner reactive with a neo-epitope formed by cleavage of a protein by a proteinase and measuring the extent of binding of peptide fragments to the immunological binding partner to measure therein protein fragments comprising the neo-epitope. Neo-epitopes from, collagen type I, collagen type III, collagen type IV, collagen type V, collagen type VI, elastin, biglycan, decorin, lumican, versican, C-reactive protein, ApoE and laminins are described.

Abstract: A method of bioassay for the quantification of peptide fragments comprising a neo-epitope formed by cleavage of a protein of an atherosclerotic plaque such as lumican, versican, perlecan, decorin, biglycan, collagen type III, CRP, ApoE, or elastin, by a proteinase, said comprises contacting a sample such as urine or serum with an antibody reactive with the neo-epitope and determining the level of binding of said immunological binding partner to peptide fragments in said sample. The assay is predictive of risk of cardiovascular disease events.

Abstract: An antibody specifically reactive with an epitope of collagen type X alpha 1 comprised in the NC1 domain C-terminal amino acid sequence SFSGFLVAPM-COOH (SEQ ID NO: 1), and a method of immunoassay for detecting in a biological sample an epitope comprised in the NC1 domain C-terminal amino acid sequence SFSGFLVAPM-COOH (SEQ ID NO: 1) of collagen type X alpha 1, by contacting said biological sample with said antibody, and determining the amount of binding of said antibody.

Abstract: Methods of diagnosis or of quantitation of fibrosis are provided that comprise conducting an immunoassay to measure neo-epitope containing protein fragments naturally present in a biofluid sample, and associating an elevation of the measure in the patient above a normal level with the presence or extent of fibrosis. The immunoassay is conducted by a contacting protein fragments naturally present in the sample with an immunological binding partner reactive with a neo-epitope formed by cleavage of a protein by a proteinase and measuring the extent of binding of peptide fragments to the immunological binding partner to measure therein protein fragments comprising the neo-epitope. The protein is collagen type III, collagen type I, collagen type IV, collagen type V, elastin, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, vimentin, or C-reactive protein.

Abstract: Provided is an antibody which binds to an epitope of collagen type VI alpha 1 comprised in the N-terminal globular domain internal amino acid sequence -ADWGQSRDAEEAISQ- (SEQ ID NO: 1). Also provided is a method of immunoassay for detecting in a biological sample an epitope comprised in the N-terminal globular domain internal amino acid sequence -ADWGQSRDAEEAISQ- (SEQ ID NO: 1) of collagen type VI alpha 1, by contacting the sample with the antibody, and determining the amount of binding of the antibody.

Abstract: Provided is a monoclonal antibody specifically reactive with a C-terminal neo-epitope of PIIINP comprised in a C-terminal amino acid sequence CPTGXQNYSP-COOH (SEQ ID NO: 4) in which X is Gly or Pro, and where the monoclonal antibody does not recognise or bind an elongated version of the C-terminal amino acid sequence CPTGXQNYSPQZ-COOH (SEQ ID NO: 5), in which Z is absent or is one or more amino acids of the sequence of collagen type III. Also provided is a method of immunoassay for detecting in a biological sample the C-terminal neo-epitope of PIIINP generated by N-protease cleavage of intact type III procollagen, by contacting the sample with the monoclonal antibody, and determining the amount of binding of the antibody.

Abstract: Provided herein are methods of diagnosis or of quantitation of fibrosis. An immunoassay is conducted to measure neo-epitope containing protein fragments of collagen type III, collagen type I, collagen type IV, collagen type V, or collagen type VI, elastin, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, vimentin, or C-reactive protein naturally present in a biofluid sample obtained from a patient. An above normal elevation of the measured protein fragments in the patient is associated with the presence or extent of fibrosis.

Abstract: An assay for citrullinated fragments of SOCS-2, Alpha 1 anti tyrpsin, versican, biglycan, laminin, or other protein having a terminal antibody binding site comprising citrulline in a blood derived sample shows diagnostic relevance in relation to rheumatoid arthritis or fibrotic disease.