# Patent

## Title
Fibrosis biomarker assay

## 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.

## Background
CROSS REFERENCE TO RELATED APPLICATIONS
This application is a continuation of pending application U.S. Ser. No. 14/960,713, filed Dec. 7, 2015, which is a divisional of U.S. Ser. No. 12/749,652, filed Mar. 30, 2013, now U.S. Pat. No. 9,206,464, which is a continuation-in-part of international application PCT/EP2008/064946, filed Nov. 4, 2008, which claims Convention priority to GB0802814.4 filed in the United Kingdom on Feb. 15, 2008, to GB0722748.1 filed in the United Kingdom on Nov. 20, 2007 and to GB0721713.6 filed in the United Kingdom on Nov. 5, 2007. The entire contents of each of the aforementioned patent applications are incorporated herein by reference.


BACKGROUND OF THE INVENTION
Field of the Invention
The present invention relates to assays for 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. 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.
Description of Related Art
Fibrotic diseases (including those listed in Table 1) are a leading cause of morbidity and mortality, e.g. cirrhosis with 800,000 death per year worldwide1.







TABLE 1







Different fibrotic diseases2








Tissue
Examples of Causes





Liver
Viral hepatitis, Schistosomiasis, Steatohepatitis



(Alcoholic or non-alcoholic)


Lung
Idiopathic pulmonary fibrosis (IPF), Systemic sclerosis



(Scleroderma)


Kidney
Nephrogenic systemic fibrosis (NSF), Diabetes,



Untreated hypertension


Heart
Heart attack, Hypertension, Atherosclerosis, Restenosis


Eye
Macular degeneration, retinal and vitreal retinopathy


Skin
Systemic sclerosis and scleroderma, keloids, hypertrophic



scars, burns, genetic factors, NFS


Pancreas
Autoimmune/hereditary causes


Intestine
Crohn's disease/inflammatory bowl disease


Brain
Alzheimer's disease, AIDS


Bone marrow
Cancer, ageing


Multi-organ
Surgical complications, chemotherapeutic drug-induced


fibrosis
fibrosis, radiation-induced fibrosis, mechanical injuries









A ‘fibrotic disease’ is any disease giving rise to fibrosis, whether as a main or a secondary symptom. Fibrosis is the end result of chronic inflammatory reactions induced by a variety of stimuli including persistent infections, autoimmune reactions, allergic responses, chemical insults, radiation, and tissue injury. Fibrosis is characterized by the accumulation and reorganization of the extracellular matrix (ECM). Despite having obvious etiological and clinical distinctions, most chronic fibrotic disorders have in common a persistent irritant that sustains the production of growth factors, proteolytic enzymes, angiogenic factors, and fibrogenic cytokines, which together stimulate the deposition of connective tissue elements, especially collagens and proteoglycans, which progressively remodel and destroy normal tissue architecture3,4. Despite its enormous impact on human health, there are currently no approved treatments that directly target the mechanisms of fibrosis5. The key cellular mediator of fibrosis is the myofibroblast, which when activated serves as the primary collagen-producing cell.
Extracellular Matrix (ECM)
Fibrogenesis is a dynamic process involving complex cellular and molecular mechanisms that usually originates from tissue injury6. Fibrogenesis is the result of an imbalance in normal ECM regulation that alters the concentration of macromolecules leading to increased tissue size and density, with progressively impaired function. These macromolecules are mainly fibrous proteins with structural and adhesive functions, such as collagens and proteoglycans.
Collagen
Collagens are widely distributed in the human body, i.e.  ̃30% of the protein mass in the human body is composed of collagens. Collagens are responsible for the structural integrity of the ECM of most connective tissues. The ECM content results from a fine balance between synthesis and degradation tightly controlled through regulation of gene expression and protein secretion, but also through endogenous protease inhibition and protein degradation by metalloproteinases and cysteine proteases7-9. Table 2 lists the major collagen types with their major tissue distribution.







TABLE 2







Major collagen types and their tissue distribution.










Collagen




type
Tissue distribution







I
Most connective tissues



II
Cartilage, vitreous humor



III
Extensible connective tissues, e.g. liver, skin,




lung, vascular system



IV
Basement membranes



V
Tissues containing collagen I



VI
Most connective tissues



VII
Skin, bladder, oral mucosa, umbilical cord, amnion



VIII
Many tissues, especially endothelium



XIII
Endothelial cells, skin, eye, heart, skeletal muscle



XIV
Vessel, bone, skin, cartilage, eye, nerve, tendon,




uterus



XXI
Vessel, heart, stomach, kidney, skeletal muscle,




placenta










Type I collagen is the most abundant collagen and is found in most connective tissues. It is especially important for the structure of bone and skin where the major collagenous components are type I and III collagens10. Collagen type I and III are the major components of liver and lung in a 1:1 ratio in healthy tissue. In addition, collagen type IV and VI are found in the basement membranes in most tissues. The most common localization of type V collagen is within the characteristic collagen fibrils, in association with the collagen type I and III10. Some collagens have a restricted tissue distribution: for example, type II, which is found almost exclusively in cartilage11. During fibrogenesis the net amount of collagens increases12-14.
Table 3 shows by way of example the collagen increase during liver fibrosis.







TABLE 3







Changes of the composition of collagen from normal to cirrhotic human liver15.















Collagen
Collagen

Distribution
Distribution


Collagen

normal liver
cirrhotic
Times
normal liver
cirrhotic


type
Chains
(mg/g)
liver (mg/g)
increased
(%)
liver (%)
















I
α1(I) α2(I)
2
16
8
37
42


III
α1(III)
2
8
4
37
21


IV
α1(IV)
0.5
7
14
9
18



α2(IV)


V
α1(V)
0.9
7
8
17
18



α2(V)



α3(V)


VI
α1(VI)
0.01
0.1
10
0.2
0.3



α2(VI)









Elastin

Elastin is a protein present in many connective tissues, primarily those that are elastic. It has a very high content of the amino acids glycine, valine, alanine, and proline, and has a molecular weight of 64 to 66 kDa. It is organised in an irregular or random coil conformation made up of 830 amino acids. Elastin is made by linking many soluble tropoelastin protein molecules, in a reaction catalyzed by lysyl oxidase, to make a massive insoluble, durable cross-linked array.
Elastin serves an important function in arteries as a medium for pressure wave propagation to help blood flow and is particularly abundant in large elastic blood vessels such as the aorta. Elastin is also very important in the lungs, elastic ligaments and the skin. Despite much efforts devoted to the understanding of elastin synthesis and turnover, neo-epitopes originating from the proteolytic cleavage of this matrix molecules have until now not been associated with disease development in fibrosis.
Vimentin
Vimentin is a member of the intermediate filament family of proteins. Intermediate filaments are an important structural feature of eukaryotic cells. They, along with microtubules and actin microfilaments, make up the cytoskeleton. Although most intermediate filaments are stable structures, in fibroblasts, vimentin exists as a dynamic structure. This filament is used as a marker for mesodermally derived tissues, and as such has been used used as an immunohistochemical marker for sarcomas.
Hertig and coworkers (Hertig et al., J Am Soc Nephrol. 2008 August; 19(8):1584-91) investigated if epithelial-to-mesenchymal transition in renal tubular epithelial cells of subjects with chronic allograft nephropathy could predict the progression of fibrosis in the allograft and measured vimentin expression in 83 biopsies from these. They did find an association between elevated vimentin expression and the intestinal fibrosis score at 1 year after surgery. In another study of hepatic fibrosis, Meriden and colleagues (Meriden et al., Clin Gastro & Hepatol 2010; 8:289-296) found a significant association between vimentin expression (in biopsies obtained at F0 stage) and fibrosis progression, with elevated levels predicting rapid progression of the hepatic fibrosis. Accordingly, we wanted to investigate if circulating fragments of vimentin could serve as sensitive and specific biomarkers of fibrosis.
Proteoglycans
Proteoglycans are a diverse group of macromolecules, which covalently link a variable number of glycosaminoglycan (GAG) side chains to a core protein16. These GAGs are polymers of disaccharide repeats (e.g. N-acetyl glucosamine or N-acetyl galactosamine), which are acidic (negatively charged) due to hydroxyl, carboxylated and sulfated side groups on the disaccharide units. This makes them highly hydrophilic, thus aiding the diffusion of water and positive ions (e.g. sodium from extracellular fluids)17. Furthermore, GAGs have the ability to form non-covalent links with for example hyaluronic acid chains to form even larger molecular complexes16. Table 4 lists the most studied proteoglycans associated with connective tissue.







TABLE 4







Proteoglycans of the extracellular matrix of connective tissue










Group
Proteoglycans
Origin
Function





Large extracellular
Aggrecan (18)
Articular cartilage
Extracellular


proteoglycans

chondrocytes,
matrix stability


(aggregating and

intervertebral
(hyaluronan


hyaluronan-binding)

disc, nasal cartilage
binding)



Versican (19, 20)
Connective tissue:
Cell-cell and




fibroblast,
cell-matrix




keratinocytes,
interactions




smooth muscle
Binding of sugars




cells, mesangial
in Ca-dependent




cells
manner



Neurocan (20)
Nervous tissue
Binds to neural





cell adhesion





molecules



Brevican (22)
Nervous tissue
Extracellular





matrix stability


Small Leucine-rich
Decorin (23)
Connective tissue,
Binds to and


proteoglycans

cartilage, bone
connect collagen


(collagen-binding)


molecules (matrix





stabilization and





thickness)





Organogenesis





Binding of TGFβ



Biglycans 24
Capillary endothelium,
Cell differentiation




skin (keratinocytes),
Binds and connect




epithelium of kidney
collagen fibrils



Fibromodulin 17
Connective tissue,
Regulate




bone, cartilage
orientation of





collagen fibers



Lumican 23
Cornea, muscle,
Controls spacing




cartilage, kidney,
and thickness of




lung, intestine
collagen fibers


Cell-associated
Serglycins 25
Widely distributed
Hemopoietic cell


proteoglycans

to endothelium -
differentiation




intercellular
Adhesion and




compartments
activation of





lymphoid cells



Syndecans 26
Widely distributed -
Binds collagens,




often cell
fibronectin,




membrane bound
thrombospondin,





tenascin and bFGF



Betaglycan 27
Widely distributed
TGFβ receptor and





signaling





Possible reservoir





of TGFβ


Basement
Perlecan 28
All basement
Selective barrier for


membrane

membranes
macromolecules


proteoglycans


Cell-adhesion









C-Reactive Protein

C-reactive protein (CRP) is an acute phase serum protein produced by the liver in response to different clinical conditions such as, inflammation, infection, or trauma29. The production of CRP is induced by cytokines such as IL-6, released from the affected or damaged tissues. The physiological role of CRP is yet unknown and discussions on its pro- or anti-inflammatory actions are ongoing.
Proteases
The imbalance between synthesis and degradation of ECM during fibrogenesis, results from conversion of the low-density subendothelial matrix into matrix rich in interstitial collagens. The increase in collagen and proteoglycans may be due to one or both of (1) a decrease in protein production and (2) impaired protein degradation, and hence less matrix degradation. The decreased protein degradation has recently received increased attention. In the regulation of this process matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs) play important roles, as well as other proteases and their inhibitors, such as cystein proteases and the cystatins.
MMPs
MMPs are a large group of endopeptidases, capable of degrading most if not all components of the ECM. Presently, more than 25 MMPs have been found. MMPs are characterized by an active site containing a metal atom, typically zinc, and are secreted as zymogens. Different MMPs are expressed in different tissues. In Table 5 MMPs in the liver are shown.







TABLE 5







MMPs in the liver30-32










Family
Protease
Source
Substrate





Collagenases
MMP-1
HSC
I, II, III, VII, VIII, X,





gelatin



MMP-8
Neutrophil
I, II, III, V, VII, X,





gelatin



MMP-13
HSC,
I, II, III, VII, X, gelatin




MFB, KC


Stromelysins
MMP-3
HSC
III, IV, V, IX, X, XI,





gelatin, laminin,





fibronectin, proteoglycans,





glycoproteins, elastin, pro-





MMP-1/13



MMP-10
HSC
III, IV, V, gelatin,





elastin, aggrecan



MMP-11
HC
PAI-1, week activity





against matrix proteins


Gelatinases
MMP-2
HSC, MBF
I, II, III, IV, V, VII, X,





XI, gelagin, elastin,





laminin



MMP-9
KC,
I, II, III, IV, V, VII, X,




HSC, HC
XI, gelagin, elastin,





laminin



MMP-7
HSC
Entactin, gelatin, elastin,





fibronectin, vitronectin,





laminin, fibrinogen


Metalloelastase
MMP-12
Macrophages
Elastin, gelatins, IV,





laminin, fibronectin,





entactin, vitronectin,





proteoglycan, myelin basic





protein, α1-antitripsin


MT-MMPs
MMP-14
HSC,
I, II, III, gelatin,




MFB, KC
fibronectin, vitronectin,





laminin, fibrinogen, pro-





MMP-2, pro-MMP-13



MMP-15
HC, BDEC
Pro-MMP-2, fibronectin,





tenascin, laminin, aggrecan,





perlecan









TIMPs block MMPs' proteolytic activity by binding in a substrate- and tissue-specific manner to MMP and membrane-type 1 metalloproteinase in a trimolecular complex (Table 6). During fibrosis TIMP levels increase dramatically, and MMP levels increase modestly or remain relatively static (except MMP-2) which in all gives a decrease in degradation of collagens.







TABLE 6







TIMPs in the liver31









Name
Sources
Metalloproteinase inhibited





TIMP-1
HSC, MFB, KC, HC
Pro-MMP-9, MMP-1, MMP-2,




MMP-3, MMP-13


TIMP-2
KC, HSC
MT-MMP-1, MT-MMP-2, proMMP-2,




MMP-3, MMP-13, MMP-7


TIMP-3
HC
MT-MMP-1, MT-MMP-2, TACE,




MMP-13









Fibroblast Activation Protein

Fibroblast Activation Protein alpha subunit (FAPa or FAP, alpha) is an integral membrane gelatinase belonging to the serine protease family. FAPa is the alpha subunit and DPP4 (CD26) the beta subunit of a heterodimeric membrane-bound proteinase complex also known as 170 kDa Melanoma Membrane Gelatinase, Integral Membrane Serine Proteinase and Seprase. Some cells make only FAPa homodimers, some only DPP4 homodimers. The monomer is inactive. FAP, alpha is selectively expressed in reactive stromal fibroblasts of epithelial cancers, granulation tissue of healing wounds, and malignant cells of bone and soft tissue sarcomas33. This protein is thought to be involved in the control of fibroblast growth or epithelial-mesenchymal interactions during development, tissue repair, and epithelial carcinogenesis. It has been shown that expression of FAP increase with the stage of fibrosis34,35.
Fibrosis Biomarkers
A number of biochemical markers have been suggested for fibrotic diseases, although not specific product of the disease. In Table 7 is an example of biochemical markers of liver fibrosis used in clinical trial. In addition there are a lot of examples of biomarkers of other fibrotic diseases12, 35-42.
Table 7 summarizes some of the known markers of liver fibrosis.















Chronic





liver


Biomarker
Parameters
disease
Reference







One





parameter


CRP

NASH
43


Hyaluronan

HCV
44-47


IGF-I

HCV
48


Leptin

HCV
49


PIIIP

HCV
50


Several


parameters


MP3
PIIINP, MMP1
HCV
51, 52


Zheng et al
HA, PIIICP, PIIINP,
Chronic
53


index
Laminin, C-IV
hepatitis


Lebensztjen
Laminin-2, C-IV, MMP2,
HBV
54


et al index
MMP9-TIMP1 index



Tenascin, hyaluronana,
HBV
55



Colalegn VI, TIMP-1


Tsochatzis et
Leptin, adiponectin,
HCV, HBC,
56


al index
resistin
NASH


Patel et al
Hyaluronan, TIMP-1, α2-
HCV
57


index
macroglobulin



TIMP-1, tenascin, collagen
NASH
58



IV, PIIINP, MMP2, laminin,



Hyaluronan


Forns-index
Age, platelet count, γGT,
HCV
51, 59-62


(76, 77)
cholesterol
HIV/HCV


FibroTest
Haptoglobin, α2-
HCV
45, 51, 60,


(76, 78)
macroglobulin,
HIV/HCV
61, 63-75



apolipoprotein A1, γGT,
NAFLD



bilirubin
NAFLD in




diabetes




patients


Actitest
FibroTest + ALT
HCV
65, 76-78


APRI (Wai-
AST, platelet count
HIV/HCV
45, 51, 60,


index)

HCV
61, 64, 66,




NAFLD
79-87


Hepascore
Bilirubin, γGT,
HCV
51, 61, 64,



hyaluronan, α2-
HIV/HCV
66, 88



macroglobulin, age, gender


FIB-4
Platelet count, AST, ALT,
HIV/HCV
61, 83



age


SHASTA
Hyaluronan, albumin, AST
HIV/HCV
61


Fibroindex
FORN + APRI
HCV
89


Fibrometer
Platelet count,
HIV/HCV
51, 61, 64,


test
prothrombin index, AST, α2-
HCV
66, 81



macroglobulin, hyaluronan,
NAFLD



urea, age


NFSA
Age, hyperglycaemia, body
NAFLD
81



mass index, platelets,



albumin, AST/ALT


Ultrasound +

HCV
82


APRI


Metwally et
Platelet count, albumin,
HCV
90


al index
AST, history of blood



transfusion, HBV core



antibody


Mohamadnejad
Age, HBV DNA levels,
HCV
91


et al index
alkaline phosphatase,



albumin, platelet counts,



AST


FibroSpect II
Hyaluronan, TIMP-1, α2-
HCV
85, 92, 93



macroglobulin


Stepwise
Combination of APRI and
HCV
94


combination
Fibrotest


algorithms


Imbert-Bismut
α2 macroglobulin, AST, ALT
HCV
95


index
γGT, total bilirubin,



albumin, α1 globulin, α2



globulin, β globulin, γ



globulin, apolipoprotein A1


Nunes et al
Age, Platelets, INR, CD4,
HCV/HIV
96



AST/ALT, Hyaluronan, YKL-
HCV



40, PIIINP


Fibroscan+++
Fibroscan, Fibrotest, APRI,
HCV
97









U.S. Pat. No. 5,387,504 describes the neo-epitope VDIPEN released by the action of stromelysin at the aggrecan site N341-F342 and an RIA assay employing a monoclonal antibody specific for this neo-epitope. More generally the use of monospecific antibodies specific for fragments of aggrecan, generated by specific stromelysin cleavage are described. Elevations of stromelysin occur in osteoarthritis, rheumatoid arthritis, atherosclerotic lesions, gout, inflammatory bowel disease (IBD), idiopathic pulmonary fibrosis (IPF), certain cancers, joint injuries, and numerous inflammatory diseases. Stromelysin is reported to be elevated in idiopathic pulmonary fibrosis, and it is alleged that the assay can be conducted on blood or other biological fluids to detect stromelysin cleavage products of aggrecan and that quantitation of such fragments can be used diagnostically in respect of IPF as well as other conditions. However, no evidence for this is provided and there have to our knowledge been no subsequent publications validating this prediction. Such RIA assays have been commercially available for many years and no reports of their successful use in diagnosing or monitoring any fibrotic disease have appeared.
U.S. Pat. No. 7,225,080 discloses a method for diagnosis of an inflammatory, a fibrotic or a cancerous disease in a patient by measuring the values of at least four biochemical markers selected from the group consisting of α2-macroglobulin, AST (aspartate aminotransferase), ALT (alanine aminotransferase), GGT (gammaglutamyl transpeptidase), γ-globulin, total bilirubin, albumin, α1-globulin, α2-globulin, haptoglobin, β-globulin, apoA1, IL-10, TGF-β1, apoA2, and apoB in the serum or plasma of said patient, and subsequently combining said values in order to determine the presence of liver fibrosis and/or liver necroinflammatory lesions in said patient. The patent does not teach the quantitative measurement of peptide fragment carrying neo-epitopes generated during fibrotic disease.
U.S. Pat. No. 6,060,255 describes a method for diagnosing the degree of liver fibrosis, comprising the steps of measuring the concentration of type IV collagen high molecular weight form in a sample using an antibody that specifically binds to type IV collagen, and relating the measurement to the degree of liver fibrosis. Again, no use is made of neo-epitopes produced by proteolytic enzymes acting in the body. The sample is actually digested with pepsin, which may obscure the natural pattern of collagen cleavage in the sample.
U.S. Pat. No. 4,628,027 (Gay) discloses the production of antibodies specific for connective tissue proteins and, more particularly, the production of monoclonal antibodies by fused cell hybrids against human collagens and enzymes involved in collagen degradation. The use of monoclonal antibodies against connective tissue proteins to establish the collagen profile of histological, cytological and biological fluid samples is described. However, the patent does not describe the measurement of connective tissue proteins based on the binding of antibodies to neo-epitopes on said connective tissue proteins.
Guañabens N et al, J Bone Miner Res, 199898 evaluated the bone turnover markers N-telopeptide of type I collagen (NTX), C-telopeptide of type I collagen (CTX) and N-terminal pro-peptide of collagen type I (PINP) in patients with primary biliary cirrhosis, a disease with increased hepatic fibrosis. The level of NTX, CTX and PINP were elevated in patients compared to controls and correlated with the histological stage of the disease. The antibodies employed in the NTX were raised against a cathepsin K cleaved site in the N-terminal of collagen type I and are dependent on the neoepitope JYDGKGVG↓(SEQ ID NO2249). The antibodies employed in the CTX were raised against a cathepsin K cleaved site in the C-terminal of collagen type I and are dependent on the neoepitope EKAHDGGR↓(SEQ ID NO2250). These markers are located in telopeptides of collagen type I and not in the internal part (the triple helical part) of collagen type I. The monoclonal antibodies employed for the PINP assay were raised against an internal epitope in the PINP sequence which is not a neo-epitope.
Møller S et al, Gut., 199999 demonstrated that the C-terminal cross linked telopeptide of type I collagen (ICTP) was elevated in alcoholic cirrhosis patients compared to controls. The study described showed that a biochemical marker can reflect hepatic fibrosis. The ICTP polyclonal antibody has been raised against trypsin and collagenase cleaved collagen type I. However, the antibodies are not binding to a neo-epitope.
Rosen H N et al, Calcif Tissue Int, 2004100 assessed the bone turnover markers N-telopeptide of type I collagen (NTX) and C-telopeptide of type I collagen (CTX) in women receiving hormone replacement treatment (HRT). In the study it was observed that the bone turnover markers decreased with treatment. The antibodies employed in the NTX were raised against a cathepsin K cleaved site in the N-terminal of collagen type I and are dependent on the neoepitope JYDGKGVG↓. The antibodies employed in the CTX were raised against a cathepsin K cleaved site in the C-terminal of collagen type I and are dependent on the neoepitope EKAHDGGR↓. In contrast to the present invention, these antibodies were used for evaluation of bone metabolism and not fibrosis.
Lein M et al, Eur Urol, 2007101 evaluated the use of the neo-epitope specific bone turnover markers N-telopeptide of type I collagen (NTX) and C-telopeptide of type I collagen (CTX) in prostate cancer patients receiving zoledronic acid. In the study it was observed that the bone turnover markers decreased with treatment. The antibodies employed in the NTX were raised against a cathepsin K cleaved site in the N-terminal of collagen type I and are dependent on the neoepitope JYDGKGVG↓. The antibodies employed in the CTX were raised against a cathepsin K cleaved site in the C-terminal of collagen type I and are dependent on the neoepitope EKAHDGGR↓. In contrast to the present invention, these antibodies were used for evaluation of the bone metabolism during invasion of bone metastases and not fibrosis.
PIIINP has been used in a number of studies to assess the severity of fibrotic disease102 in patients with skin fibrosis following severe burn trauma103, for disease progression in noncirrhotic primary biliary cirrhosis104, in primary biliary cirrhosis and chronic viral hepatitis C105. PIIINP and ICTP were measured in patients with fibrosis of the myocardium106.
Many reports combine a set of biochemical markers to improve the predictive value of the biochemical index. Eleven different serum markers were measured in 205 patients with fibrotic staging from F0 to F4, and the most informative markers were alpha2 macroglobulin, alpha2 globulin (or haptoglobin), gamma globulin, apolipoprotein A1, gamma glutamyltranspeptidase, and total bilirubin107. An index of these markers had a negative predictive value (100% certainty of absence of F2, F3, or F4) was obtained for scores ranging from zero to 0.10 (12% [41] of all patients), and high positive predictive value (>90% certainty of presence of F2, F3, or F4) for scores ranging from 0.60 to 1.00 (34% [115] of all patients). However, in none of the above mentioned reports is it suggested that measurements of peptide fragments based on antibodies binding to neo-epitopes as now claimed might be useful for the assessment of patients with fibrotic disease.
SUMMARY OF THE INVENTION
The present invention now provides a method of diagnosis of fibrosis comprising, conducting an immunoassay to measure neo-epitope containing protein fragments naturally present in a patient biofluid sample, and associating an elevation of said measure in said patient above a normal level with the presence of fibrosis, wherein said immunoassay is conducted by a method comprising: contacting protein fragments naturally present in said 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 said immunological binding partner to measure therein protein fragments comprising said neo-epitope, and wherein said protein is collagen type I, collagen type III, collagen type IV, collagen type V or collagen type VI, biglycan, decorin, lumican, versican, perlecan, neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, CRP, or vimentin subject to the proviso that when the neo-epitopes are formed by cleavage of type I collagen, the cleavage is not at a site at which collagen type I is cleaved by cathepsin K. WO2009/059972 published on 14th May 2009 (after the priority date hereof) discloses assays for neo-epitopes of collagen III, but does not disclose that an elevated level of such a measure is to be associated with the presence or extent of fibrosis. Optionally, an assay according to this invention is based on one of the proteins named above other than collagen Type III or if based on collagen Type III utilises an immunological binding partner against one of the neoepitopes formed at the cleavage sites PGIPGRNGDP* SEQ ID NO1, *ESCPTGPQNY SEQ ID NO2, or PKGDTGPRGP* SEQ ID NO3 (where * marks the cleavage site).
For these purposes, cardiovascular disease may not be regarded as fibrosis, or the fibrosis detected according to the invention may be other than fibrosis accompanying cardiovascular disease. Optionally, an elevated result in an immunoassay according to this invention is associated with skin fibrosis, lung fibrosis, or liver fibrosis.
The method may comprise the preliminary step of obtaining a patient biofluid sample.
The invention includes a method of immunoassay to measure neo-epitope containing protein fragments naturally present in body fluid sample, wherein said immunoassay is conducted by a method comprising: contacting protein fragments naturally present in said 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 said immunological binding partner to measure therein protein fragments comprising said neo-epitope, and wherein said protein is neurocan, brevican, fibromodulin, serglycin, syndecan, betaglycan, collagen type I, collagen type IV, collagen type V, collagen type VI, CRP, or vimentin subject to the proviso that when the neo-epitopes are formed by cleavage of type I collagen, the cleavage is not at a site at which collagen type I is cleaved by cathepsin K.
Optionally, an assay according to this invention is based on one of the proteins named above other than collagen Type III or if based on collagen Type III utilises an immunological binding partner against one of the neoepitopes formed at the cleavage sites PGIPGRNGDP* SEQ ID NO1, *ESCPTGPQNY SEQ ID NO2, or PKGDTGPRGP* SEQ ID NO3 (where * marks the cleavage site).
Said immunological binding partner may have specific binding affinity for peptide fragments comprising a C-terminal neoepitope or an N-terminal neoepitope.
Specific reactivity with or immunological affinity for a neo-epitope will imply that the relevant immunological binding partner is not reactive with intact protein from which the neo-epitope derives. Preferably, said immunological binding partner is not reactive with a neo-epitope sequence, such as a sequence listed below, if the sequence is prolonged past the respective cleavage site.
The term ‘immunological binding partner’ as used herein includes polyclonal and monoclonal antibodies and also specific binding fragments of antibodies such as Fab or F(ab′)2. Thus, said immunological binding partner may be a monoclonal antibody or a fragment of a monoclonal antibody having specific binding affinity.
Preferably, said peptide fragments are fragments of Type I, III, IV, V, or VI collagen, elastin, C-reactive protein, or one of the proteoglycans Biglycan, Decorin, Versican, and Perlecan. The connective tissue proteins are preferred. Preferably, the neo-epitope sequence to which the immunological binding partner binds is not found in any other protein or is not found in any of the other proteins to which the method of the invention relates.
Several candidate proteases may be responsible for the digestion of proteins in the fibrotic tissues. Most likely, this is the result of the large range of complicated processes resulting in different neo-epitope profiles dependent on the levels of disease.



BRIEF DESCRIPTION OF THE SEVERAL VIEWS OF THE DRAWINGS
The invention will be further described and illustrated with reference to the following examples and the accompanying drawings.
FIG. 1 shows a graph showing CO3 ELISA results of different biological samples: Pooled human serum samples (Serum); Human amniotic fluid (AF); Human fibroblast culture media (Fibr. Cltr.).
FIGS. 2A-2B show a graph showing CO3 serum levels in sham operated (s) and bile duct ligated rats at baseline (b) and at termination (t) (FIG. 2A) and the corresponding delta-values of CO3 in rat serum: Termination levels-Baseline levels (FIG. 2B).
FIG. 3 shows a graph showing CO3 levels in different human serum samples. Normal serum: from healthy individuals. COPD: Chronic Obstructed Pulmonary Disease (leading to lung fibrosis). Scleroderma (leading to skin and lung fibrosis). HCV: Hepatitis virus C (leading to liver fibrosis);
FIGS. 4A-4C show liver weights (FIG. 4A) and liver scores (FIG. 4B) and Sirius Red photomicrographs showing the hepatic structure in sham-operated rats, and in BDL rats 2 and 4 weeks post-surgery determined in Example 5.
FIGS. 5A-5C show levels of MMP-9 cleavage fragments of Type III collagen measured according to the invention in Example 5. FIG. 5A shows MMP-9 mediated CO3 degradation serum levels in bile duct ligated BDL- or sham-operated rats. FIG. 5B shows CO3-610C delta values (termination-baseline paired) 2 weeks post-surgery P<0.0001 and 4 weeks post-surgery P=0.0016. FIG. 5C shows ctx-II levels in BDL- or sham-operated rats.
FIG. 6 shows levels of Type III collagen gene expression in BDL or sham-operated rats determined in Example 5.
FIGS. 7A-7B show changes of expression levels of the MMP-9 cleavage fragment of Type III collagen reactive with the antibody used in Example 5 as determined by Western blot. FIG. 7A shows Western blot 2 and 4 weeks post-surgery and FIG. 7B shows bands from Western Blot quantified by densitometry.
FIGS. 8A-8B show the results of histology staining of liver sections obtained in Example 5. FIG. 8A shows histology sections from BDL- or sham-operated rats stained with Sirius Red (top) and masked histology secions for quantifying total collagen content in the liver (bottom). FIG. 8B shows total collagen quantified by Visiopharm software 2 weeks post-surgery P=0.0081 and 4 weeks post-surgery P=0.0047.
FIGS. 9A-9C shows correlations between measurements of fragments of Type III collagen according to the invention with other liver biomarkers as determined in Example 5. FIG. 9A shows a correlation of Col3al to CO3-610C was found. FIG. 9B shows a correlation of CO3-610C to % collagen was found. FIG. 9C shows that a correlation of Col3al to % collagen was found.
FIG. 10 shows results obtained on human serum samples in Example 6.
FIG. 11 shows results obtained in testing the reactivity of a monoclonal antibody recognising an N-terminal neo-epitope from CRP.
FIG. 12 shows collagen accumulation in rat liver measured in Example 8.
FIG. 13 shows immunoassay results obtained in Example 8.
FIG. 14 shows the correlation of the immunoassay results of FIG. 13 with liver collagen content.
FIG. 15 shows a comparison of the results of an immunoassay according to the invention with measurements of hyaluronic acid and of Sirius red staining in Example 8.
FIGS. 16A-16B show the correlation of results from the immunoassay according to the invention with Sirius red staining (FIG. 16A) and the correlation between hyaluronic acid levels and Sirius red staining (FIG. 16B).
FIG. 17 shows the lack of correlation between the results of the immunoassay of the invention and hyaluronic acid levels.
FIGS. 18A-18D show skin sections and skin thickness measurements described in Example 9. FIG. 18A shows a skin section from a PBS treated mouse at 8 weeks of treatment. FIG. 18B shows a skin section section from Bleomycin treated mouse at 8 weeks of treatment. FIGS. 18C-18D are plots of skin thickness increase between P/bs (n=7/time point) and Bleomycin (n=13/time point) treated mice for 2 weeks (P=0.0029), 4 weeks (P=0.0004), 6 weeks (P<0.0001) and 8 weeks ((P<0.0001).
FIG. 19 shows results from an immunoassay according to the invention in Example 9.
FIGS. 20A-20B shows Western Blot images obtained in Example 9 (FIG. 20A) and the corresponding immunoassay results (FIG. 20B).
FIG. 21 shows a correlation between immunoassay results and skin thickness measurements.
FIG. 22 shows a correlation between urine immunoassay results and Western blot measurements described in Example 9.



DETAILED DESCRIPTION OF THE INVENTION
Collagen Assays
Collagen Type I
We have determined that the enzymes listed in the following table cleave type I collagen at least the following cleavage sites (marked “.”):







TABLE 8







Collagen type I cleavage sites.








Protease
Collagen type I





MMP-2
V.PGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO: 4


 


MMP-2
S.VPGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO: 5


 


MMP-2
G.ISVPGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO: 6


 


MMP-9
G.ISVPGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO: 6


 


MMP-13
G.FQGPPGEPGEPGASGPMGPRGPPGPPG.K SEQ ID NO: 7


 


MMP-13
V.PGPMGPSGPRGLPGPPGAPGPQG.F SEQ ID NO: 8


 


MMP-2
F.SGLDGAKGDAGPAGPKGEPGSPGENGAPGQMGPRG.L SEQ ID NO: 9


 


MMP-9
F.SGLDGAKGDAGPAGPKGEPGSPGENGAPGQMGPRG.L SEQ ID NO: 9


 


MMP-13
F.SGLDGAKGDAGPAGPKGEPGSPGENGAPGQMGPRG.L SEQ ID NO: 9


 


MMP-9
G.LPGERGRPGAPGPAG.A SEQ ID NO: 10


 


MMP-13
G.LPGERGRPGAPGPAG.A SEQ ID NO: 10


 


MMP-2
G.LTGSPGSPGPDGKTGPPGPAG.Q SEQ ID NO: 11


 


MMP-2
E.RGSPGPAGPKGSPGEAGRPGEAGLPGAKG.L SEQ ID NO: 12


 


MMP-2
G.ERGSPGPAGPKGSPGEAGRPGEAGLPGAKG.L SEQ ID NO: 13


 


MMP-9
G.LTGSPGSPGPDGKTGPPGPAG.Q SEQ ID NO: 14


 


MMP-9
G.LTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPPG.A SEQ ID NO: 15


 


MMP-9
G.LTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPPGARG.Q SEQ ID NO: 16


 


MMP-13
G.LTGSPGSPGPDGKTGPPGPAG.Q SEQ ID NO: 14


 


MMP-13
G.ERGSPGPAGPKGSPGEAGRPGEAGLPGAKG.L SEQ ID NO: 13


 


MMP-9
G.QDGRPGPPGPPGARG.Q SEQ ID NO: 17


 


MMP-9
G.LTGSPGSPGPDGKTGPPGPAGQDGRPGPPGPPGARG.Q SEQ ID NO: 18


 


MMP-2
G.KDGEAGAQGPPGPAGPAGERGEQGPAGSPGF.Q SEQ ID NO: 19


 


MMP-2
G.ERGEQGPAGSPGF.Q SEQ ID NO: 20


 


MMP-3
E.RGVPGPPGAVGPAGKDGEAGAQGPPGPAGPAGERGEQGPAGSPGF.Q



SEQ ID NO: 21


 


MMP-8
E.RGVPGPPGAVGPAGKDGEAGAQGPPGPAGPAGERGEQGPAGSPGF.Q



SEQ ID NO: 21


 


—
113 PKGDTGPRGP.122 SEQ ID NO: 22









P indicates hydroxyproline, M indicates oxidised methionine, and K indicates hydroxylysine.
The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type I collagen, excluding cleavage at a cathepsin K type I collagen site. Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:







TABLE 9





N-terminal sequences of protease generated peptide


fragments of Collagen type I.


(The symbol ‘.’ Indicates the cleavage site)







Collagen I, alpha1









.ISVPGP SEQ ID NO: 23
.VPGPMG SEQ ID NO: 24
.PGPMGP SEQ ID NO: 25


 


.LPGPGG SEQ ID NO: 26
.FQGPPG SEQ ID NO: 27
.KNGDDG SEQ ID NO: 28


 


.ARGLPG SEQ ID NO: 29
.SGLDGA SEQ ID NO: 30
.LDGAKG SEQ ID NO: 31


 


.LPGERG SEQ ID NO: 32
.VRGEPG SEQ ID NO: 33
.PGAKGA SEQ ID NO: 34


 


.GGPPGP SEQ ID NO: 35
.NSGEPG SEQ ID NO: 36
.DGVAGP SEQ ID NO: 37


 


.ERGSPG SEQ ID NO: 38
.LTGSPG SEQ ID NO: 39
.QDGRPG SEQ ID NO: 40


 


.RGVPGP SEQ ID NO: 41
.VGPAGK SEQ ID NO: 42
.ERGEQG SEQ ID NO: 43


 


.RGEQGP SEQ ID NO: 44
.PGERGV SEQ ID NO: 45
.ANGAPG SEQ ID NO: 46


 


.ARGAPG SEQ ID NO: 47
.PGDRGE SEQ ID NO: 48
.AKGDAG SEQ ID NO: 49


 


.PIGNVG SEQ ID NO: 50
.AAGRVG SEQ ID NO: 51
.PPGPAG SEQ ID NO: 52


 


.GADGPA SEQ ID NO: 53
.GPQGIA SEQ ID NO: 54
.GQRGVV SEQ ID NO: 55


 


.QRGVVG SEQ ID NO: 56
.GLPGQR SEQ ID NO: 57
.PGLPGP SEQ ID NO: 58


 


.PMGPPG SEQ ID NO: 59
.MGPPGL SEQ ID NO: 60
.DKGETG SEQ ID NO: 61


 


.LQGPPG SEQ ID NO: 62
.SAGAPG SEQ ID NO: 63
.RTGDAG SEQ ID NO: 64


 


.FDFSF SEQ ID NO: 65
.DFSF SEQ ID NO: 66
.ATGAAG SEQ ID NO: 67


 


.AKGEAG SEQ ID NO: 68
.GIAGAP SEQ ID NO: 69
.IAGAPG SEQ ID NO: 70


 


.VQGPPG SEQ ID NO: 71
.LPGPPG SEQ ID NO: 72
.AGPKGS SEQ ID NO: 73


 


.RGSPGP SEQ ID NO: 74
.FAGPPG SEQ ID NO: 75
.QAGVMG SEQ ID NO: 76


 


.ARGQAG SEQ ID NO: 77
.NVGAPG SEQ ID NO: 78
.PAGERG SEQ ID NO: 79


 


.KDGEAG SEQ ID NO: 80
.GEVGPP SEQ ID NO: 81
.ARGERG SEQ ID NO: 82


 


.QGLPGP SEQ ID NO: 83
.IAGQRG SEQ ID NO: 84
.LTGPIG SEQ ID NO: 85


 


.AGLPGP SEQ ID NO: 86
.RGVVGL SEQ ID NO: 87
.AGPPGA SEQ ID NO: 88


 


.LAGPPG SEQ ID NO: 89
.EPGKQG SEQ ID NO: 90
.ATGFPG SEQ ID NO: 91


 


.PSGASG SEQ ID NO: 92
.GKQGPS SEQ ID NO: 93
.GPPGPA SEQ ID NO: 94


 


.AGQRGV SEQ ID NO: 95
.ARGPAG SEQ ID NO: 96
.ASGPAG SEQ ID NO: 97


 


.VVGLPG SEQ ID NO: 98
.VGPPGP SEQ ID NO: 99
.GPPGPP SEQ ID NO: 100


 


.TGDAGP SEQ ID NO: 175









Alternatively, suitable immunological binding partners may be specifically reactive with any of the following sequences at the C terminal of a peptide:







TABLE 10





C-terminal sequences of protease generated peptide


fragments of Collagen type I


(The symbol ‘.’ Indicates the cleavage site).







Collagen I, alpha1









QLSYGY. SEQ ID NO: 101
EKSTGG. SEQ ID NO: 102
PPGPQG. SEQ ID NO: 103


 


KGHRGF. SEQ ID NO: 104
PSGPRG. SEQ ID NO: 105
APGPQG. SEQ ID NO: 106


 


APGPAG. SEQ ID NO: 107
FPGAVG. SEQ ID NO: 108
SEGPQG. SEQ ID NO: 109


 


GANGAP. SEQ ID NO: 110
ANGAPG. SEQ ID NO: 46
SGPQGP. SEQ ID NO: 112


 


EPGPVG. SEQ ID NO: 113
EPGPTG. SEQ ID NO: 114
RGFPGA. SEQ ID NO: 115


 


KGPAGE. SEQ ID NO: 116
RGSPGP. SEQ ID NO: 74
LPGAKG. SEQ ID NO: 118


 


PPGPPG. SEQ ID NO: 119
PPGARG. SEQ ID NO: 120
PGKAGE. SEQ ID NO: 121


 


AVGPAG. SEQ ID NO: 122
PAGPAG. SEQ ID NO: 123
AGPAGE. SEQ ID NO: 124


 


APGPDG. SEQ ID NO: 125
RGERGF. SEQ ID NO: 126
PAGPRG. SEQ ID NO: 127


 


KDGVRG. SEQ ID NO: 128
PAGPTG. SEQ ID NO: 129
TGARGA. SEQ ID NO: 130


 


PGPAGF. SEQ ID NO: 131
EPGDAG. SEQ ID NO: 132
PAGPPG. SEQ ID NO: 133


 


SAGPPG. SEQ ID NO: 134
ATGFPG. SEQ ID NO: 91
NAGPPG. SEQ ID NO: 136


 


GEVGPP. SEQ ID NO: 81
GEKGSP. SEQ ID NO: 138
GAPGTP. SEQ ID NO: 139


 


PGPQGI. SEQ ID NO: 140
GPQGIA. SEQ ID NO: 54
PQGIAG. SEQ ID NO: 142


 


IAGQRG. SEQ ID NO: 84
GQRGVV. SEQ ID NO: 55


 


GPSGEP. SEQ ID NO: 146
ERGPPG. SEQ ID NO: 147
RGPPGP. SEQ ID NO: 148


 


PVGPVG. SEQ ID NO: 149
PQGPRG. SEQ ID NO: 150
HRGFSG. SEQ ID NO: 151


 


EQGPSG. SEQ ID NO: 152
PRGPPG. SEQ ID NO: 153
PPGPRG. SEQ ID NO: 154


 


GPPGPP. SEQ ID NO: 100
GPPSAG. SEQ ID NO: 156
PPSAGF. SEQ ID NO: 157


 


PPGPAG. SEQ ID NO: 52
TPGPQG. SEQ ID NO: 160


 


QMGPRG. SEQ ID NO: 161
PGPPGA. SEQ ID NO: 162
QGIAGQ. SEQ ID NO: 163


 


PGADGQ. SEQ ID NO: 164
AGSPGF. SEQ ID NO: 165
LPGPSG. SEQ ID NO: 166


 


PPGPKG. SEQ ID NO: 167
PGERGA. SEQ ID NO: 168
PMGPPG. SEQ ID NO: 59


 


PKGPAG. SEQ ID NO: 170
GRNGDP. SEQ ID NO: 171
SPGEQG. SEQ ID NO: 172


 


GPAGRP. SEQ ID NO: 173
PPGPIG. SEQ ID NO: 174
TGDAGP. SEQ ID NO: 175


 


TGPRGP. SEQ ID NO: 177









Collagen Type III

We have determined that the enzymes listed in the following table cleave type Ill collagen at least the following cleavage sites (marked *):







TABLE 11







Cleavage sites in collagen type III.








Protease
Neo-Epitope





MMP-1
A*GIPGAPGLMGARGPPGPA*G SEQ ID NO: 178


 


MMP-1
K*GDPGPPGIPGRNGDPGI*P SEQ ID NO: 179


 


MMP-1
G*LAGPPGMPGPRGSPGPQG*V SEQ ID NO: 180


 


MMP-1
G*ERGLPGPPGIKGPAGIPGF*P SEQ ID NO: 181


 


MMP-1
G*IAGITGARGLAGPPGMPGPR*G SEQ ID NO: 182


 


MMP-1
G*IKGHRGFPGNPGAPGSPGPAG*Q SEQ ID NO: 183


 


MMP-1
A*RGLAGPPGMPGPRGSPGPQGV*K SEQ ID NO: 184


 


MMP-1
I*TGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 185


 


MMP-1
I*TGARGLAGPPGMPGPRGSPGPQGV*K SEQ ID NO: 186


 


MMP-1
G*ITGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 187


 


MMP-1
G*VKGESGKPGANGLSGERGPPGPQG*L SEQ ID NO: 188


 


MMP-1
G*SRGAPGPQGPRGDKGETGERGAAG*I SEQ ID NO: 189


 


MMP-1
P*KGDAGQPGEKGSPGAQGPPGAPGPLG*I SEQ ID NO: 190


 


MMP-1
G*ITGARGLAGPPGMPGPRGSPGPQGV*K SEQ ID NO: 191


 


MMP-1
G*LRGGAGPPGPEGGKGAAGPPGPPGAAGTPG*L SEQ ID NO: 192


 


MMP-1
G*HAGAQGPPGPPGINGSPGGKGEMGPAGIPGAPG*L SEQ ID NO: 193


 


MMP-1
A*GKSGDRGESGPAGPAGAPGPAGSRGAPGPQGPRGDKGETGERGAAG*I



SEQ ID NO: 194


 


MMP-1
G*LQGLPGTGGPPGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG*L



SEQ ID NO: 195


 


MMP-3
G*ERGLPGPPGIKGPAGIPGF*P SEQ ID NO: 196


 


MMP-3
A*VGGLAGYPGPAGPPGPPGPPGT*S SEQ ID NO: 197


 


MMP-3
K*DGTSGHPGPIGPPGPRGNRGER*G SEQ ID NO: 198


 


MMP-3
A*VGGLAGYPGPAGPPGPPGPPGTSGHPG*S SEQ ID NO: 199


 


MMP-3
G*IAGITGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 200


 


MMP-3
A*PGAPGGKGDAGAPGERGPPGLAGAPGLRG*G SEQ ID NO: 201


 


MMP-3
A*VGGLAGYPGPAGPPGPPGPPGTSGHPGSPG*S SEQ ID NO: 202


 


MMP-2
A*IGSPGPAGPRGPVGPSGPPG*K SEQ ID NO: 203


 


MMP-3 & -8
G*AIGSPGPAGPRGPVGPSGPPG*K SEQ ID NO: 204


 


MMP-8
P*AGQQGAIGSPGPA*G SEQ ID NO: 205


 


MMP-8
G*GPPGVAGPPGGSGPAGPP*G SEQ ID NO: 206


 


MMP-8
L*AGPPGMPGPRGSPGPQG*V SEQ ID NO: 207


 


MMP-8
G*LSGERGPPGPQGLPGLA*G SEQ ID NO: 208


 


MMP-8
R*GLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 209


 


MMP-8
G*LAGPPGMPGPRGSPGPQGV*K SEQ ID NO: 210


 


MMP-8
R*GLAGPPGMPGPRGSPGPQGV*K SEQ ID NO: 211


 


MMP-8
G*PQGPPGKNGETGPQGPPGP*T SEQ ID NO: 212


 


MMP-8
G*VKGERGSPGGPGAAGFPGAR*G SEQ ID NO: 213


 


MMP-8
A*RGLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 214


 


MMP-8
N*GLSGERGPPGPQGLPGLAGTA*G SEQ ID NO: 215


 


MMP-8
A*VGGLAGYPGPAGPPGPPGPPGT*S SEQ ID NO: 216


 


MMP-8
G*SPGGKGEMGPAGIPGAPGLMGA*R SEQ ID NO: 217


 


MMP-8
T*GARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 218


 


MMP-8
V*KGESGKPGANGLSGERGPPGPQG*L SEQ ID NO: 219


 


MMP-8
G*VKGERGSPGGPGAAGFPGARGLPGPPGSNGNPGPPGPSGSPGKDGPPGPAG*N



SEQ ID NO: 220


 


MMP-8
G*SPGAQGPPGAPGPLGIAGITGARGLAGPPG*M SEQ ID NO: 221


 


MMP-8
R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQ*G SEQ ID NO: 222


 


MMP-8
R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQ*G SEQ ID NO: 223


 


MMP-8
G*IAGITGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 224


 


MMP-9
G*IKGPAGIPGFPG*M SEQ ID NO: 225


 


MMP-9
G*QPGVMGFPGPKG*N SEQ ID NO: 226


 


MMP-9
G*IKGPAGIPGFPGMK*G SEQ ID NO: 227


 


MMP-9
G*IKGPAGIPGFPGMKG*H SEQ ID NO: 228


 


MMP-9
I*PGAPGLMGARGPPGPAG*A SEQ ID NO: 229


 


MMP-9
G*ERGLPGPPGIKGPAGIP*G SEQ ID NO: 230


 


MMP-9
G*IPGAPGLMGARGPPGPAG*A SEQ ID NO: 231


 


MMP-9
G*FRGPAGPNGIPGEKGPAG*E SEQ ID NO: 232


 


MMP-9
P*GIPGQPGSPGSPGPPGIC*E SEQ ID NO: 233


 


MMP-9
G*ERGLPGPPGIKGPAGIPGF*P SEQ ID NO: 234


 


MMP-9
A*VGGLAGYPGPAGPPGPPGPPG*T SEQ ID NO: 235


 


MMP-9
G*VKGERGSPGGPGAAGFPGARG*L SEQ ID NO: 236


 


MMP-9
G*DAGAPGAPGGKGDAGAPGERGPPG*L SEQ ID NO: 237


 


MMP-9
Q*GPPGPTGPGGDKGDTGPPGPQGL*Q SEQ ID NO: 238


 


MMP-9
G*INGSPGGKGEMGPAGIPGAPGLM*G SEQ ID NO: 239


 


MMP-9
Q*GPPGEPGQAGPSGPPGPPGAIGPS*G SEQ ID NO: 240


 


MMP-9
P*GPPGINGSPGGKGEMGPAGIPGAP*G SEQ ID NO: 241


 


MMP-9
R*GLPGPPGSNGNPGPPGPSGSPGKDGPPGPAG*N SEQ ID NO: 242


 


MMP-9
G*KNGETGPQGPPGPTGPGGDKGDTGPPGPQG*L SEQ ID NO: 243


 


MMP-9
G*LPGIAGPRGSPGERGETGPPGPAGFPGAPG*Q SEQ ID NO: 244


 


MMP-9
G*INGSPGGKGEMGPAGIPGAPGLMGARGPPGPAG*A SEQ ID NO: 245


 


MMP-9
P*GINGSPGGKGEMGPAGIPGAPGLMGARGPPGPAG*A SEQ ID NO: 246


 


MMP-9
P*PGENGKPGEPGPKGDAGAPGAPGGKGDAGAPGERGPPG*L SEQ ID NO: 247


 


MMP-9
G*LKGENGLPGENGAPGPMGPRGAPGERGRPGLPGAAG*A SEQ ID NO: 248


 


MMP-9
G*NTGAPGSPGVSGPKGDAGQPGEKGSPGAQGPPGAPGPLG*I



SEQ ID NO: 249


 


MMP-9
G*LMGARGPPGPAGANGAPGLRGGAGEPGKNGAKGEPGPRG*E SEQ ID NO: 250


 


MMP-9
G*LRGGAGPPGPEGGKGAAGPPGPPGAAGTPGLQGMPGERGGLGSPGPKG*D



SEQ ID NO: 251


 


MMP-8 & -9
G*QQGAIGSPGPAGPRGPVGPSGPPG*K SEQ ID NO: 252


 


MMP-9
K*GDPGPPGIPGRNGDPGIPGQPG*S SEQ ID NO: 253


 


MMP-9
G*LRGGAGPPGPEGGKGAAGPPGPPG*A SEQ ID NO: 254


 


MMP-9
G*KNGETGPQGPPGPTGPGGDKGDTGPPGPQG*L SEQ ID NO: 255


 


MMP-9
G*YQGPPGEPGQAGPSGPPGPPG*A SEQ ID NO: 256


 


MMP-9
G*VAGPPGGSGPAGPPGPQG*V SEQ ID NO: 257


 


MMP-8, -9 & -
G*DKGEPGGPGADGVPGKDGPRGPTGPIGPPGPAG*Q SEQ ID NO: 258


13


 


ADAMTS-5
Q*GHAGAQGPPGPPGIN*G SEQ ID NO: 259


 


CathepsinK
A*GERGAPGPA*G SEQ ID NO: 260


 


CathepsinK
A*GIPGFPGMK*G SEQ ID NO: 261


 


CathepsinK
F*PGMKGHRGFD*G SEQ ID NO: 262


 


CathepsinK
G*FPGARGLPGPPG*S SEQ ID NO: 263


 


CathepsinK
A*GFPGARGLPGPPG*S SEQ ID NO: 264


 


CathepsinK
P*PGPPGPPGTSGHP*G SEQ ID NO: 265


 


CathepsinK
G*FPGMKGHRGFD*G SEQ ID NO: 266


 


CathepsinK
Q*PGDKGEGGAPGLPGI*A SEQ ID NO: 267


 


CathepsinK
R*GDKGETGERGAAGIK*G SEQ ID NO: 268


 


CathepsinK
D*GRNGEKGETGAPGLK*G SEQ ID NO: 269


 


CathepsinK
A*GQPGDKGEGGAPGLPGIA*G SEQ ID NO: 270


 


CathepsinK
G*GPPGENGKPGEPGPKGD*A SEQ ID NO: 271


 


CathepsinK
A*GIPGFPGMKGHRGFD*G SEQ ID NO: 272


 


CathepsinK
R*GGAGEPGKNGAKGEPGPR*G SEQ ID NO: 273


 


CathepsinK
K*GERGSPGGPGAAGFPGARGLPGPP*G SEQ ID NO: 274


 


CathepsinK
EDGKDGSPGEPGANGLP*G SEQ ID NO: 275


 


CathepsinK
G*AAGFPGARGLPGPPGSNGNPGPPGPS*G SEQ ID NO: 276


 


CathepsinK
R*PGPPGPSGPRGQPGVMGFPGPKGN*D SEQ ID NO: 277


 


CathepsinK
Q*GPPGPPGINGSPGGKGEMGPAGIPGAP*G SEQ ID NO: 278


 


CathepsinK
A*GKDGESGRPGRPGERGLPGPPGIK*G SEQ ID NO: 279


 


CathepsinK
A*GARGNDGARGSDGQPGPPGPPGTAGFPG*S SEQ ID NO: 280


 


CathepsinK
S*PGVSGPKGDAGQPGEKGSPGAQGPPGAPG*P SEQ ID NO: 281


 


CathepsinK
R*GSDGQPGPPGPPGTAGFPGSPGAKGEVGPA*G SEQ ID NO: 282


 


CathepsinK
Q*GPPGPPGINGSPGGKGEMGPAGIPGAPGLM*G SEQ ID NO: 283


 


CathepsinK
A*GPPGPPGPPGTSGHPGSPGSPGYQGPPGEPG*Q SEQ ID NO: 284


 


CathepsinK
F*PGAPGQNGEPGGKGERGAPGEKGEGGPPGVA*G SEQ ID NO: 285


 


CathepsinK
A*GFPGAPGQNGEPGGKGERGAPGEKGEGGPPG*V SEQ ID NO: 286


 


CathepsinK
A*GARGNDGARGSDGQPGPPGPPGTAGFPGSPGAKGEVGPA*G



SEQ ID NO: 287


 


CathepsinK
R*GAAGEPGRDGVPGGPGMRGMPGSPGGPGSDGKPGPPGSQGESGRPGPPGPS*G



SEQ ID NO: 288


 


CathepsinS
G*IAGITGARGL*A SEQ ID NO: 289


 


CathepsinS
A*GPPGPPGAAGTPGLQG*M SEQ ID NO: 290


 


CathepsinS
N*GLSGERGPPGPQGLPG*L SEQ ID NO: 291


 


CathepsinS
M*GARGPPGPAGANGAPGLR*G SEQ ID NO: 292


 


CathepsinS
N*GLSGERGPPGPQGLPGLA*G SEQ ID NO: 293


 


CathepsinS
G*IAGITGARGLAGPPGMPGPRG*S SEQ ID NO: 294


 


CathepsinS
G*IAGITGARGLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 295


 


CathepsinS
R*GGAGPPGPEGGKGAAGPPGPPGAAGTPGLQ*G SEQ ID NO: 296


 


CathepsinS
S*GPKGDAGQPGEKGSPGAQGPPGAPGPLG*I SEQ ID NO: 297


 


CathepsinS
G*IAGITGARGLAGPPGMPGPRGSPGPQGVK*G SEQ ID NO: 298


 


CathepsinS
S*GPKGDAGQPGEKGSPGAQGPPGAPGPLG*I SEQ ID NO: 299


 


CathepsinS
G*IAGITGARGLAGPPGMPGPRGSPGPQGVK*G SEQ ID NO: 300


 


CathepsinS
A*VGGLAGYPGPAGPPGPPGPPGTSGHPGSPGSPGYQ*G SEQ ID NO: 301


 


CathepsinS
E*PGPQGHAGAQGPPGPPGINGSPGGKGEMGPAGIPGAPG*L SEQ ID NO: 302


 


ADAMTS1
I*PGFPGMKGHR*G SEQ ID NO: 303


 


ADAMTS1
R*GSPGGPGAAGFPGAR*G SEQ ID NO: 304


 


ADAMTS1
K*GPAGIPGFPGMKGHR*G SEQ ID NO: 305


 


ADAMTS1
R*GLAGPPGMPGPRGSPGPQ*G SEQ ID NO: 306


 


ADAMTS1
A*GITGARGLAGPPGMPGPR*G SEQ ID NO: 307


 


ADAMTS1
L*GIAGITGARGLAGPPGMPGPR*G SEQ ID NO: 308


 


ADAMTS1
T*GARGLAGPPGMPGPRGSPGPQ*G SEQ ID NO: 309


 


ADAMTS1
Q*GPPGPPGINGSPGGKGEMGPAG*I SEQ ID NO: 310


 


ADAMTS1
L*PGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO: 311


 


ADAMTS1
A*GITGARGLAGPPGMPGPRGSPGPQ*G SEQ ID NO: 312


 


ADAMTS1
T*GARGLAGPPGMPGPRGSPGPQGVK*G SEQ ID NO: 313


 


ADAMTS1
R*GLPGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO: 314


 


ADAMTS1
G*RPGLPGAAGARGNDGARGSDGQPGPPG*P SEQ ID NO: 315


 


ADAMTS1
N*GAPGPMGPRGAPGERGRPGLPGAAGAR*G SEQ ID NO: 316


 


ADAMTS1
A*GSRGAPGPQGPRGDKGETGERGAAGIK*G SEQ ID NO: 317


 


ADAMTS1
R*GLAGPPGMPGPRGSPGPQGVKGESGKPGAN*G SEQ ID NO: 318


 


ADAMTS1
R*GLAGPPGMPGPRGSPGPQGVKGESGKPGANGL*S SEQ ID NO: 319


 


ADAMTS1
P*GPPGSNGNPGPPGPSGSPGKDGPPGPAGNTGAPGS*P SEQ ID NO: 320


 


ADAMTS1
T*GARGLAGPPGMPGPRGSPGPQGVKGESGKPGAN*G SEQ ID NO: 321


 


ADAMTS1
R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQGVKGER*G SEQ ID NO: 322


 


ADAMTS1
G*GPPGVAGPPGGSGPAGPPGPQGVKGERGSPGGPGAAGF*P SEQ ID NO: 323


 


ADAMTS1
K*SGDRGESGPAGPAGAPGPAGSRGAPGPQGPRGDKGETGERGAAGIK*G



SEQ ID NO: 324


 


ADAMTS4
I*PGFPGMKGHR*G SEQ ID NO: 325


 


ADAMTS4
R*GLAGPPGMPGPR*G SEQ ID NO: 326


 


ADAMTS4
G*PQGLQGLPGTGGPP*G SEQ ID NO: 327


 


ADAMTS4
K*GPAGIPGFPGMKGHR*G SEQ ID NO: 328


 


ADAMTS4
R*GLAGPPGMPGPRGSPGPQG*V SEQ ID NO: 329


 


ADAMTS4
G*GPPGENGKPGEPGPKGDAGAP*G SEQ ID NO: 330


 


ADAMTS4
A*PGFRGPAGPNGIPGEKGPAGER*G SEQ ID NO: 331


 


ADAMTS4
E*KGSPGAQGPPGAPGPLGIAGITGAR*G SEQ ID NO: 332


 


ADAMTS4
L*PGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO: 333


 


ADAMTS4
R*GAPGFRGPAGPNGIPGEKGPAGER*G SEQ ID NO: 334


 


ADAMTS4
R*GLPGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO: 335


 


ADAMTS4
R*GPVGPSGPPGKDGTSGHPGPIGPPGPR*G SEQ ID NO: 336


 


ADAMTS4
A*PGPQGPRGDKGETGERGAAGIKGHR*G SEQ ID NO: 337


 


ADAMTS4
R*GAPGPQGPRGDKGETGERGAAGIKGHR*G SEQ ID NO: 338


 


ADAMTS4
R*GFPGNPGAPGSPGPAGQQGAIGSPGPAGPR*G SEQ ID NO: 339


 


ADAMTS4
L*PGPPGIKGPAGIPGFPGMKGHRGFDGR*N SEQ ID NO: 340


 


ADAMTS4
D*AGQPGEKGSPGAQGPPGAPGPLGIAGITGAR*G SEQ ID NO: 341


 


ADAMTS4
R*GPTGPIGPPGPAGQPGDKGEGGAPGLPGIAGPR*G SEQ ID NO: 342


 


ADAMTS4
K*GDAGQPGEKGSPGAQGPPGAPGPLGIAGITGAR*G SEQ ID NO: 343


 


ADAMTS4
R*NGEKGETGAPGLKGENGLPGENGAPGPMGPR*G SEQ ID NO: 344


 


ADAMTS4
A*PGFRGPAGPNGIPGEKGPAGERGAPGPAGPRGA*A SEQ ID NO: 345


 


ADAMTS4
R*GAPGFRGPAGPNGIPGEKGPAGERGAPGPAGPR*G SEQ ID NO: 346


 


ADAMTS4
R*GSPGERGETGPPGPAGFPGAPGQNGEPGGKGER*G SEQ ID NO: 347


 


ADAMTS4
G*HAGAQGPPGPPGINGSPGGKGEMGPAGIPGAPGLMG*A SEQ ID NO: 348


 


ADAMTS4
R*GLAGPPGMPGPRGSPGPQGVKGESGKPGANGLSGER*G SEQ ID NO: 349


 


ADAMTS8
L*GIAGITGARGL*A SEQ ID NO: 350


 


ADAMTS8
I*PGFPGMKGHR*G SEQ ID NO: 351


 


ADAMTS8
R*GLAGPPGMPGPR*G SEQ ID NO: 352


 


ADAMTS8
Q*GPPGAPGPLGIAGITGAR*G SEQ ID NO: 353


 


ADAMTS8
A*GITGARGLAGPPGMPGPR*G SEQ ID NO: 354


 


ADAMTS8
A*GIPGAPGLMGARGPPGPAGAN*G SEQ ID NO: 355


 


ADAMTS8
R*GLAGPPGMPGPRGSPGPQGVKG*E SEQ ID NO: 356


 


ADAMTS8
K*GSPGAQGPPGAPGPLGIAGITGAR*G SEQ ID NO: 357


 


ADAMTS8
L*PGPPGIKGPAGIPGFPGMKGHR*G SEQ ID NO: 358


 


ADAMTS8
K*DGTSGHPGPIGPPGPRGNRGER*G SEQ ID NO: 359


 


ADAMTS8
A*GITGARGLAGPPGMPGPRGSPGPQ*G SEQ ID NO: 360


 


ADAMTS8
R*GLAGPPGMPGPRGSPGPQGVKGESG*K SEQ ID NO: 361


 


ADAMTS8
R*GLAGPPGMPGPRGSPGPQGVKGESGKPGAN*G SEQ ID NO: 362


 


ADAMTS8
R*GLAGPPGMPGPRGSPGPQGVKGESGKPGANGL*S SEQ ID NO: 363


 


ADAMTS8
P*GPPGSNGNPGPPGPSGSPGKDGPPGPAGNTGAPGS*P SEQ ID NO: 364


 


ADAMTS8
R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQGVKGER*G SEQ ID NO: 365


 


ADAMTS8
K*SGDRGESGPAGPAGAPGPAGSRGAPGPQGPRGDKGETGERGA*A



SEQ ID NO: 366


 


ADAMTS8
R*GAPGEKGEGGPPGVAGPPGGSGPAGPPGPQGVKGERGSPGGPGAAGFPGAR*G



SEQ ID NO: 367


 


MMP9
 *AIGPSG    * SEQ ID NO: 368


 


MMP9
117′ PGIPGRNGDP*. 124′ SEQ ID NO: 369


 


MMP9
142′ .*ESCPTGPQNY 151′ SEQ ID NO: 370


 


MMP9
113′ PKGDTGPRGP*. ′122 SEQ ID NO: 371









The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type Ill collagen. Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:







TABLE 12





N-terminal sequences of protease generated peptide


fragments of Collagen type III.







Collagen type III









GIPGAP SEQ ID NO: 372
GDPGPP SEQ ID NO: 373
LAGPPG SEQ ID NO: 89


 


IAGITG SEQ ID NO: 375
IKGHRG SEQ ID NO: 376
RGLAGP SEQ ID NO: 377


 


TGARGL SEQ ID NO: 378
ITGARG SEQ ID NO: 379
VKGESG SEQ ID NO: 380


 


KGDAGQ SEQ ID NO: 381
LRGGAG SEQ ID NO: 382
ERGLPG SEQ ID NO: 385


 


GKSGDR SEQ ID NO: 383
LQGLPG SEQ ID NO: 384
AIGSPG SEQ ID NO: 143


 


DGTSGH SEQ ID NO: 135
VGGLAG SEQ ID NO: 155
LSGERG SEQ ID NO: 176


 


GPPGVA SEQ ID NO: 158
AGPPGM SEQ ID NO: 145
IGSPGP SEQ ID NO: 386


 


GLSGER SEQ ID NO: 387
GLAGPP SEQ ID NO: 388
PQGPPG SEQ ID NO: 389


 


GARGLA SEQ ID NO: 111
KGESGK SEQ ID NO: 374
VKGERG SEQ ID NO: 159


 


GAPGEK SEQ ID NO: 141
QPGVMG SEQ ID NO: 144
IKGPAG SEQ ID NO: 169


 


IPGAPG SEQ ID NO: 117
FRGPAG SEQ ID NO: 137
QQGAIG SEQ ID NO: 390


 


GPPGPT SEQ ID NO: 391
INGSPG SEQ ID NO: 392
GPPGEP SEQ ID NO: 393


 


GLPGPP SEQ ID NO: 394
KNGETG SEQ ID NO: 395
LPGIAG SEQ ID NO: 396


 


GINGSP SEQ ID NO: 397
PGENGK SEQ ID NO: 398
LKGENG SEQ ID NO: 399


 


LMGARG SEQ ID NO: 400
YQGPPG SEQ ID NO: 401
GERGAP SEQ ID NO: 402


 


DKGEPG SEQ ID NO: 403
GHAGAQ SEQ ID NO: 404
GSDGQP SEQ ID NO: 405


 


PGMKGH SEQ ID NO: 406
FPGARG SEQ ID NO: 407
GFPGAR SEQ ID NO: 408


 


FPGMKG SEQ ID NO: 409
PGDKGE SEQ ID NO: 410
GDKGET SEQ ID NO: 411


 


GQPGDK SEQ ID NO: 412
GPPGEN SEQ ID NO: 413
GIPGFP SEQ ID NO: 414


 


GERGSP SEQ ID NO: 415
PGVPGA SEQ ID NO: 416
AAGFPG SEQ ID NO: 417


 


GPPGPP SEQ ID NO: 100
GKDGES SEQ ID NO: 418
GARGND SEQ ID NO: 419


 


GFPGAP SEQ ID NO: 420
NTGAPG SEQ ID NO: 437
GAAGEP SEQ ID NO: 421


 


IAGITG SEQ ID NO: 375
GLSGER SEQ ID NO: 387
GARGPP SEQ ID NO: 422


 


GPPGSN SEQ ID NO: 423
GPKGDA SEQ ID NO: 424
GGAGPP SEQ ID NO: 425


 


PGPQGH SEQ ID NO: 426
PGFPGM SEQ ID NO: 427
GSPGGP SEQ ID NO: 428


 


SGDRGE SEQ ID NO: 429
GITGAR SEQ ID NO: 430
GIAGIT SEQ ID NO: 431


 


PGPPGI SEQ ID NO: 432
ESCPTG SEQ ID NO: 433
HAGAQG SEQ ID NO: 434


 


GAPGFR SEQ ID NO: 435
RPGLPG SEQ ID NO: 436
GAPGPM SEQ ID NO: 438


 


GSPGER SEQ ID NO: 439
PQGLQG SEQ ID NO: 440
GPAGIP SEQ ID NO: 441


 


AIGPSG SEQ ID NO: 368
PGFRGP SEQ ID NO: 443
KGSPGA SEQ ID NO: 444


 


GAPGPQ SEQ ID NO: 445
GFPGNP SEQ ID NO: 446
GPVGPS SEQ ID NO: 447


 


GPTGPI SEQ ID NO: 448
GDAGQP SEQ ID NO: 449
NGEKGE SEQ ID NO: 450


 


SRGAPG SEQ ID NO: 451
VAGPPG SEQ ID NO: 452
PGPQGP SEQ ID NO: 453


 


AGQPGE SEQ ID NO: 454
PGAPGG SEQ ID NO: 455
PGAPGQ SEQ ID NO: 456


 


AGQQGA SEQ ID NO: 457
PGPPGP SEQ ID NO: 458
SPGGKG SEQ ID NO: 459


 


GARGLA SEQ ID NO: 111
GRNGEK SEQ ID NO: 460
GPPGAP SEQ ID NO: 461


 


GSRGAP SEQ ID NO: 462
GGAGEP SEQ ID NO: 463
GSPGAQ SEQ ID NO: 464


 


SPGAQG SEQ ID NO: 465
PGVSGP SEQ ID NO: 466
PGAPGL SEQ ID NO: 467


 


GIPGQP SEQ ID NO: 468
DAGAPG SEQ ID NO: 469
GPPGIN SEQ ID NO: 470









or with any of the following sequences at the C-terminal of a peptide:








TABLE 13





C-terminal sequences of protease generated peptide


fragments of Collagen type III.







Collagen type III









GPPGPA SEQ ID NO: 94
NGDPGI SEQ ID NO: 471
SPGPQG SEQ ID NO: 472


 


GMPGPR SEQ ID NO: 473
SPGPAG SEQ ID NO: 474
PGPQGV SEQ ID NO: 475


 


ERGAAG SEQ ID NO: 476
PGPLGI SEQ ID NO: 477
AAGTPG SEQ ID NO: 478


 


ERGPPG SEQ ID NO: 147
PGPPGT SEQ ID NO: 479
GNRGER SEQ ID NO: 480


 



APGLRG SEQ ID NO: 481
HPGSPG SEQ ID NO: 482


 


PSGPPG SEQ ID NO: 483
GSPGPA SEQ ID NO: 484
GPAGPP SEQ ID NO: 485


 


GLPGLA SEQ ID NO: 486
GPPGPQ SEQ ID NO: 490
QGPPGP SEQ ID NO: 487


 


GLAGTA SEQ ID NO: 488
FPGPKG SEQ ID NO: 491
PGLMGA SEQ ID NO: 489


 


LAGPPG SEQ ID NO: 89
GPAGIP SEQ ID NO: 441
FPGARG SEQ ID NO: 407


 


IPGFPG SEQ ID NO: 492
PPGPPG SEQ ID NO: 119
GFPGMK SEQ ID NO: 493


 


FPGAPG SEQ ID NO: 494
GAIGPS SEQ ID NO: 495
AGIPGF SEQ ID NO: 496


 


GPPGIC SEQ ID NO: 2187
LPGAAG SEQ ID NO: 2188
APGPLG SEQ ID NO: 2189


 


PGPQGL SEQ ID NO: 497
GAPGLM SEQ ID NO: 498
GPPGIN SEQ ID NO: 470


 


SPGPKG SEQ ID NO: 499
GEPGPR SEQ ID NO: 500
IPGQPG SEQ ID NO: 501


 


TGAPGS SEQ ID NO: 502
GHRGFD SEQ ID NO: 503
LPGPPG SEQ ID NO: 72


 


PGPKGD SEQ ID NO: 506
PGLPGI SEQ ID NO: 504
GAAGIK SEQ ID NO: 505


 


GLPGIA SEQ ID NO: 507
PQGLPG SEQ ID NO: 508
GAPGLR SEQ ID NO: 509


 


GLPGPP SEQ ID NO: 394
GANGLP SEQ ID NO: 510
GPPGPS SEQ ID NO: 511


 


IPGAPG SEQ ID NO: 117
GPPGIK SEQ ID NO: 512
TAGFPG SEQ ID NO: 513


 


GEVGPA SEQ ID NO: 514
PPGPQG SEQ ID NO: 103
GPPGVA SEQ ID NO: 158


 


GEVGPA SEQ ID NO: 499
GFPGAR SEQ ID NO: 408
TGARGL SEQ ID NO: 378


 


EKGPAG SEQ ID NO: 515
EPGPRG SEQ ID NO: 516
PPGAPG SEQ ID NO: 517


 


TSGHPG SEQ ID NO: 518
GAPGPA SEQ ID NO: 519
TPGLQG SEQ ID NO: 520


 


GTPGLQ SEQ ID NO: 521
GTSGHP SEQ ID NO: 522
MPGPRG SEQ ID NO: 523


 


GPQGVK SEQ ID NO: 524
GAPGLK SEQ ID NO: 525
GSPGYQ SEQ ID NO: 526


 


PPGPAG SEQ ID NO: 52
PGPKGN SEQ ID NO: 527
GAAGAR SEQ ID NO: 528


 


FPGMKG SEQ ID NO: 409
PGANGL SEQ ID NO: 529
GTGGPP SEQ ID NO: 530


 


GLSGER SEQ ID NO: 387
TGPRGP SEQ ID NO: 177
GITGAR SEQ ID NO: 430


 


GMKGHR SEQ ID NO: 531
EGGPPG SEQ ID NO: 532
GSPGPQ SEQ ID NO: 533


 


EMGPAG SEQ ID NO: 534
GIAGPR SEQ ID NO: 535
QPGPPG SEQ ID NO: 536


 


GRNGDP SEQ ID NO: 171
GKPGAN SEQ ID NO: 537
VKGESG SEQ ID NO: 380


 


GVKGER SEQ ID NO: 538
PGAAGF SEQ ID NO: 539
TGERGA SEQ ID NO: 540


 


PQGVKG SEQ ID NO: 541
GDAGAP SEQ ID NO: 542
GPAGER SEQ ID NO: 543


 


GPPGPR SEQ ID NO: 544
GPAGPR SEQ ID NO: 545
RGFDGR SEQ ID NO: 546


 


AGPRGA SEQ ID NO: 547
GGKGER SEQ ID NO: 548
APGLMG SEQ ID NO: 549


 


GPAGAN SEQ ID NO: 550









Collagen IV

We have determined that the enzymes listed in the following table cleave type IV collagen at least the following cleavage sites (marked “.”):







TABLE 14







Cleavage fragments of collagen type IV








Protease
Neo-Epitope





FAP
D.IDGYRGPPGP.Q SEQ ID NO: 551


 


FAP
S.MGPPGTPSVDHGF.L SEQ ID NO: 552


 


FAP
P.DGLPGSMGPPGTPSVDHG.F SEQ ID NO: 553


 


FAP
P.DGLPGSMGPPGTPSVDHGF.L SEQ ID NO: 554


 


FAP
P.DGLPGSMGPPGTPSVDHGFL.V SEQ ID NO: 555


 


FAP
P.SGRDGLPGPPGSPGPPGQPGY.T SEQ ID NO: 556


 


FAP
P.SGRDGLPGPPGSPGPPGQPGYTN.G SEQ ID NO: 557


 


FAP
P.SGRDGLPGPPGSPGPPGQPGYTNG.I SEQ ID NO: 558


 


FAP
I.PGSKGEQGFMGPPGPQGQPGLPGS.P SEQ ID NO: 559


 


FAP
P.RGFPGPPGPDGLPGSMGPPGTPSVD.H SEQ ID NO: 560


 


FAP
E.PGPPGLPGSVGSPG.V SEQ ID NO: 561


 


FAP
I.DGYRGPPGPQGP.P SEQ ID NO: 562


 


FAP
P.RGFPGPPGPDGLPGSMG.P SEQ ID NO: 563


 


FAP
D.GLPGSMGPPGTPSVDHGF.L SEQ ID NO: 564


 


FAP
D.GLPGSMGPPGTPSVDHGFL.V SEQ ID NO: 565


 


FAP
P.GLPGQQGAPGIPGFPGSKGEMGVMGTP.G SEQ ID NO: 566


 


FAP
I.GIPGMPGSPGLKGSPGSVGYPGSPGLPGE.K SEQ ID NO: 567


 


FAP
P.GPPGPPGEKGQMGLSFQGPKGDKGDQGVSGPPGVP.G



SEQ ID NO: 568


 


FAP
P.GIGPPGARGPPGGQGPPGLSGPPGIKGEKGFPGFPGL.D



SEQ ID NO: 569


 


FAP
E.PGLPGIPGVSGPK.G SEQ ID NO: 570


 


FAP
G.EKGQKGDTGPPGPPGLV.I SEQ ID NO: 571


 


FAP
L.PGIGVQGPPGPPGIPGPIGQPGLHGIPGEKGDPGPP.G SEQ ID NO: 572


 


FAP
G.SPGIPGHQGEMG.P SEQ ID NO: 573


 


FAP
E.PGMQGEPGPPGP.P SEQ ID NO: 574


 


FAP
G.PPGRLGAPGTPGLPGP.R SEQ ID NO: 575


 


FAP
P.PGPKGFPGIPGP.P SEQ ID NO: 576


 


FAP
A.KGQPGLPGFPGT.P SEQ ID NO: 577


 


FAP
D.RGPPGPPGIRGPPGP.P SEQ ID NO: 578


 


FAP
P.GPPGEKGKPGQDGIPGP.A SEQ ID NO: 579


 


FAP
L.LGSKGEKGEPGLPGIPGVSGPKGY.Q SEQ ID NO: 580


 


MMP-9
D.GLPGSMGPPGTPSVDHG.F SEQ ID NO: 581


 


MMP-9
D.GLPGSMGPPGTPSVDHGF.L SEQ ID NO: 564


 


MMP-9
T.GPLGEKGERGYPGTPGPRGE.P SEQ ID NO: 582


 


MMP-9
G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPG.L



SEQ ID NO: 583


 


MMP-9
P.DGLPGSMGPPGTPSVDHGFL.V SEQ ID NO: 555


 


MMP-9
D.PGLKGDKGDVGLPGKPGSMDKVDMGS.M SEQ ID NO: 584


 


MMP-9
L.PGPMGPPGLPGIDGV.K SEQ ID NO: 585


 


MMP-9
D.GLPGSMGPPGTPSVDHGFL.V SEQ ID NO: 565


 


MMP-9
G.IRGEPGPPGLPGSVGSPGVPGIGPPG.A SEQ ID NO: 586


 


MMP-9
G.FPGPPGPDGLPGSMGPPGTPSVDHGF.L SEQ ID NO: 587


 


MMP-9
G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPG.A SEQ ID NO: 588


 


MMP-9
G.IRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPG.L



SEQ ID NO: 589


 


MMP-9
E.DGVIGMMGFPGAIGP.P SEQ ID NO: 590


 


MMP-9
Y.PGNPGILGPPGEDGVIGMMGFPGAIGPPGPPG.N SEQ ID NO: 591


 


MMP-9
I.PPSDEICEPGPPGP.P SEQ ID NO: 592


 


MMP-9
L.PGLPGPKGEPGLPGYPGNPGIKGS.V SEQ ID NO: 593


 


MMP-9
G.IKGDKGSMGHPGPKGPP.G SEQ ID NO: 594


 


MMP-9
T.PGSPGCAGSPGLPGSPGPPG.P SEQ ID NO: 595


 


MMP-9
P.GAPGPQGLPGPPGFPGPVGPPGPPGFFGFPGAMGPRGPKGHMGE.R



SEQ ID NO: 596


 


MMP-9
G.LPGFAGNPGP SEQ ID NO: 597


 


MMP-9 + FAP
G.AEGLPGSPGFPGPQG.D SEQ ID NO: 598


 


MMP-9 + FAP
M.GPPGVPGFQGPKGLP.G SEQ ID NO: 599


 


MMP-9 + FAP
D.IDGYRGPPGPQGPPG.E SEQ ID NO: 600


 


MMP-9 + FAP
G.DQGDQGVPGAKGLPGP.P SEQ ID NO: 601


 


MMP-9 + FAP
G.DRGPQGQPGLPGLPGP.M SEQ ID NO: 602


 


MMP-9 + FAP
P.DGLPGSMGPPGTPSVDHGF.L SEQ ID NO: 554


 


MMP-9 + FAP
E.KGSIGIPGMPGSPGLKGSPGSVGYP.G SEQ ID NO: 603


 


MMP-9 + FAP
G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPG.A SEQ ID NO: 588


 


MMP-9 + FAP
G.IRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPG.L



SEQ ID NO: 589


 


MMP-9 + FAP
G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPG.L



SEQ ID NO: 583


 


MMP-9 + FAP
G.LQGIRGEPGPPGLPGSVGSPGVPGIGPPGARGPPGGQGPPGLSGPPG.I



SEQ ID NO: 604


 


MMP-9 + FAP
I.PPSDEICEPGPPGP.P SEQ ID NO: 592


 


MMP-9 + FAP
P.GPPGLMGPPGPPGLPGP.K SEQ ID NO: 605


 


MMP-9 + FAP
G.ERGSPGIPGAPGPIGPPGSPG.L SEQ ID NO: 606


 


MMP-9 + FAP
P.GIPGAPGAPGFPGSKGEPGDILTFPGMKGDKGELGSPGAPGL.P



SEQ ID NO: 607


 


MMP-9 + FAP
C.DGGVPNTGPPGEPGPP.G SEQ ID NO: 608


 


MMP12, Alpha1
.ILGHVPGML. SEQ ID NO: 2190


 


MMP12, Alpha1
.PGLPGQPGPPGLPVPGQ. SEQ ID NO: 2191


 


MMP12, Alpha1
.SGYPGNPGLPGIPGQDGPPGPPGIPGCNGTKGERGPLGPPGL.



SEQ ID NO: 2192


 


MMP12, Alpha1
.VSGPPGVPGQA. SEQ ID NO: 2193


 


MMP12, Alpha1
.VSGPPGVPGQAQ. SEQ ID NO: 2194


 


MMP12, Alpha2
.KRGPPGPPGLPGPPGPDGFL. SEQ ID NO: 2195


 


MMP12, Alpha2
.LHGFPGAPGQEGPLG. SEQ ID NO: 2196


 


MMP12, Alpha2
.LPGPDGPPGERGLPGEVL. SEQ ID NO: 2197


 


MMP12, Alpha2
.LRGIPGF. SEQ ID NO: 2198


 


MMP12, Alpha2
.PGFPGAPGTVGAPGIAGIPQK. SEQ ID NO: 2199


 


MMP12, Alpha2
.QQGNRGLGF. SEQ ID NO: 2200


 


MMP12, Alpha2
.VGQPGPNGIPSDTL. SEQ ID NO: 2201


 


MMP12, Alpha3
.GEPGMQGEPGPPGPPGNLGPCGPRGKPGKDGKPGTPGPAGEKG.



SEQ ID NO: 2202


 


MMP12, Alpha3
.GEPGPPGPPGNLGPCGPRGKPGKDGKPGTPGPAGEKGNK.



SEQ ID NO: 2203


 


MMP12, Alpha3
.PGIPGTPGPPGLPGLQGPVGPPG. SEQ ID NO: 2204


 


MMP12, Alpha3
.PGDIVFRK. SEQ ID NO: 2205


 


MMP12, Alpha4
.GNKGDPASHFGPPGPKG. SEQ ID NO: 2206


 


MMP12, Alpha4
.PGPRGKPGM. SEQ ID NO: 2207


 


MMP12, Alpha5
.PGLPGQPGTRGL. SEQ ID NO: 2208


 


MMP12, Alpha5
.PGPPGPLGIPGRSGVPGLKGDDGLQGQPGLPGPTGEKGSK.



SEQ ID NO: 2209


 


MMP12, Alpha5
.PGPPGPLGIPGRSGVPGLKGDDGLQGQPGLPGPTGEKGSKG.



SEQ ID NO: 2210


 


MMP12, Alpha5
.SKGEKGEPGLPGIPGVSGPKGYQGLPGDPGQPGLSGQPGL.



SEQ ID NO: 2211









The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type IV collagen. Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:







TABLE 15





N-terminal sequences of protease generated peptide


fragments of Collagen type IV.







Collagen type IV









IDGYRG SEQ ID NO: 609
MGPPGT SEQ ID NO: 610
DGLPGS SEQ ID NO: 611


 


PGSKGE SEQ ID NO: 612
RGFPGP SEQ ID NO: 613
PGPPGL SEQ ID NO: 614


 


GPPGPP SEQ ID NO: 100
GLPGSM SEQ ID NO: 615
GLPGQQ SEQ ID NO: 616


 


LGSKGE SEQ ID NO: 617
GIGPPG SEQ ID NO: 611
PGLPGI SEQ ID NO: 504


 


PGIGVQ SEQ ID NO: 618
SPGIPG SEQ ID NO: 619
PGMQGE SEQ ID NO: 620


 


PGPKGF SEQ ID NO: 621
KGQPGL SEQ ID NO: 622
RGPPGP SEQ ID NO: 148


 


PPSDEI SEQ ID NO: 623
PGLKGD SEQ ID NO: 624
GPLGEK SEQ ID NO: 625


 


IRGEPG SEQ ID NO: 626
FPGPPG SEQ ID NO: 627
LQGIRG SEQ ID NO: 628


 


DGVIGM SEQ ID NO: 629
PGNPGI SEQ ID NO: 630
PGLPGP SEQ ID NO: 58


 


IKGDKG SEQ ID NO: 631
PGSPGC SEQ ID NO: 632
GAPGPQ SEQ ID NO: 445


 


GPPGVP SEQ ID NO: 633
DQGDQG SEQ ID NO: 634
DRGPQG SEQ ID NO: 442


 


KGSIGI SEQ ID NO: 635
PPGRLG SEQ ID NO: 636
EKGQKG SEQ ID NO: 637


 


ERGSPG SEQ ID NO: 38
GIPGAP SEQ ID NO: 372
DGGVPN SEQ ID NO: 638


 


SGRDGL SEQ ID NO: 639
GPPGEK SEQ ID NO: 640
AEGLPG SEQ ID NO: 641


 


DGYRGP SEQ ID NO: 642
GPPGLM SEQ ID NO: 643
LPGFAG SEQ ID NO: 644


 


GIPGMP SEQ ID NO: 645
PGPMGP SEQ ID NO: 25
.PGIPGT SEQ ID NO: 2227


 


.ILGHVP SEQ ID NO: 2212
.LPGPDG SEQ ID NO: 2214
.PGDIVF SEQ ID NO: 2215


 


.PGLPGQ SEQ ID NO: 2213
.LRGIPG SEQ ID NO: 760
.GNKGDP SEQ ID NO: 2216


 


.SGYPGN SEQ ID NO: 2217
.PGFPGA SEQ ID NO: 2218
.PGPRGK SEQ ID NO: 2219


 


.VSGPPG SEQ ID NO: 2220
.QQGNRG SEQ ID NO: 2221
.PGPPGP SEQ ID NO: 458


 


.KRGPPG SEQ ID NO: 2223
.VGQPGP SEQ ID NO: 2222
.SKGEKG SEQ ID NO: 2226


 


.LHGFPG SEQ ID NO: 2225
.GEPGMQ SEQ ID NO: 2224
.GEPGPP SEQ ID NO: 675









or with any of the following sequences at the C-terminal of a peptide:








TABLE 16





C-terminal sequences of protease generated peptide


fragments of Collagen type IV.







Collagen type IV









RGPPGP SEQ ID NO: 148
SVDHGF SEQ ID NO: 646
PSVDHG SEQ ID NO: 647


 


VDHGFL SEQ ID NO: 648
PGQPGY SEQ ID NO: 649
QPGYTN SEQ ID NO: 650


 


PGLPGS SEQ ID NO: 651
GTPSVD SEQ ID NO: 652
SVGSPG SEQ ID NO: 653


 


LPGSMG SEQ ID NO: 654
GPPGVP SEQ ID NO: 633
PGFPGL SEQ ID NO: 655


 


PGLPGE SEQ ID NO: 656
GDPGPP SEQ ID NO: 373
HQGEMG SEQ ID NO: 657


 


GPPGLV SEQ ID NO: 658
PGIPGP SEQ ID NO: 659
PGFPGT SEQ ID NO: 660


 


PGLPGP SEQ ID NO: 58
DGIPGP SEQ ID NO: 661
SGPKGY SEQ ID NO: 662


 


PGPRGE SEQ ID NO: 663
GQGPPG SEQ ID NO: 664
KVDMGS SEQ ID NO: 665


 


GIGPPG SEQ ID NO: 611
PGIDGV SEQ ID NO: 666
KGHMGE SEQ ID NO: 667


 


PGAIGP SEQ ID NO: 668
PPGPPG SEQ ID NO: 119
KGLPGP SEQ ID NO: 669


 


GPKGPP SEQ ID NO: 671
SPGPPG SEQ ID NO: 672
GPKGLP SEQ ID NO: 670


 


GSVGYP SEQ ID NO: 673
PQGPPG SEQ ID NO: 389
PPGSPG SEQ ID NO: 674


 


GEPGPP SEQ ID NO: 675
AGNPGP SEQ ID NO: 676
PGIKGS SEQ ID NO: 677


 


PGYTNG SEQ ID NO: 678
FPGPQG SEQ ID NO: 679
PGPQGP SEQ ID NO: 453


 


LSGPPG SEQ ID NO: 680
PGAPGL SEQ ID NO: 467
GVMGTP SEQ ID NO: 681


 


GVSGPK SEQ ID NO: 682
PGPPGP SEQ ID NO: 458


 


HVPGML. SEQ ID NO: 2228
LPVPGQ. SEQ ID NO: 2229
LGPPGL. SEQ ID NO: 2230


 


GVPGQA. SEQ ID NO: 2231
VPGQAQ. SEQ ID NO: 2232
GPDGFL. SEQ ID NO: 2233


 


QEGPLG. SEQ ID NO: 2234
LPGEVL. SEQ ID NO: 2235
RGIPGF. SEQ ID NO: 2236


 


NRGLGF. SEQ ID NO: 2238
IPSDTL. SEQ ID NO: 2239
PAGEKG. SEQ ID NO: 2240


 


GEKGNK. SEQ ID NO: 2241
PVGPPG. SEQ ID NO: 2242
DIVFRK. SEQ ID NO: 2243


 


PPGPKG. SEQ ID NO: 167
RGKPGM. SEQ ID NO: 2244
PGTRGL. SEQ ID NO: 2245


 


GEKGSK. SEQ ID NO: 2246
EKGSKG. SEQ ID NO: 2247
SGQPGL. SEQ ID NO: 2248


 


AGIPQK. SEQ ID NO: 2237









Collagen V

We have determined that the enzymes listed in the following table cleave type v collagen at least the following cleavage sites (marked “.” or in the ablsence of a ‘.’, at the end of the sequence):







TABLE 14A







Cleavage fragments of collagen type V








Protease
Neo-epitope (COV)





MMP2, Alpha3
K.GDPGPPGPIGSLG.H SEQ ID NO: 683


 


MMP2, Alpha3
G.LRGIPGPVGEPG.L SEQ ID NO: 684


 


MMP2, Alpha3
V.IGPPGLQGLPGPPGE.K SEQ ID NO: 685


 


MMP2, Alpha3
G.KDGIPGPLGPLGPPG.A SEQ ID NO: 686


 


MMP2, Alpha3
G.LRGIPGPVGEPGLL.G SEQ ID NO: 687


 


MMP2, Alpha3
G.VLGPQGKTGEVGPLG.E SEQ ID NO: 688


 


MMP2, Alpha3
K.DGIPGPLGPLGPPGAA.G SEQ ID NO: 689


 


MMP2, Alpha3
G.EDGERGAEGPPGPTG.Q SEQ ID NO: 690


 


MMP2, Alpha3
G.LQGPPGFPGPKGPPG.H SEQ ID NO: 691


 


MMP2, Alpha3
P.IGSLGHPGPPGVAGPLG.Q SEQ ID NO: 692


 


MMP2, Alpha3
G.IRGPPGTVIMMPFQ.F SEQ ID NO: 693


 


MMP2, Alpha3
G.QMGPPGPLGPSGLPGLK.G SEQ ID NO: 694


 


MMP2, Alpha3
G.LLGAPGQMGPPGPLGPSG.L SEQ ID NO: 695


 


MMP2, Alpha3
G.LRGIPGPVGEPGLLGAPG.Q SEQ ID NO: 696


 


MMP2, Alpha3
G.LLGPRGSPGPTGRPGVTG.I SEQ ID NO: 697


 


MMP2, Alpha3
G.IRGPPGTVIMMPFQF.A SEQ ID NO: 698


 


MMP2, Alpha3
G.KDGIPGPLGPLGPPGAAGP.S SEQ ID NO: 699


 


MMP2, Alpha3
G.KDGIPGPLGPLGPPGAAGPSG.E SEQ ID NO: 700


 


MMP2, Alpha3
Q.GLPGLEGREGAKGELGPPGPLG.K SEQ ID NO: 701


 


MMP2, Alpha3
L.GPIGEKGKSGKTGQPGLEGERGPPGSRG.E SEQ ID NO: 702


 


MMP2, Alpha3
G.LRGIPGPVGEPGLLGAPGQMGPPGPLGPSG.L SEQ ID NO: 703


 


MMP2, Alpha3
G.ANGSPGERGPLGPAGGIGLPGQSGSEGPVGPAG.K SEQ ID NO: 704


 


MMP2, Alpha3
G.LIGTPGEKGPPGNPGIPGLPGSDGPLGHPGHEGPTG.E SEQ ID NO: 705


 


MMP2, Alpha1
G.LPGEPGPRG.L SEQ ID NO: 706


 


MMP2, Alpha1
L.ALRGPAGPMG.L SEQ ID NO: 707


 


MMP2, Alpha1
R.LALRGPAGPMG.L SEQ ID NO: 708


 


MMP2, Alpha1
G.LTGRPGPVGPPGSGG.L SEQ ID NO: 709


 


MMP2, Alpha1
G.LLGPKGPPGPPGPPG.V SEQ ID NO: 710


 


MMP2, Alpha1
G.IPGRPGPQGPPGPAG.E SEQ ID NO: 711


 


MMP2, Alpha1
P.GPDGPPGPMGPPGLP.G SEQ ID NO: 712


 


MMP2, Alpha1
G.QPGPSGADGEPGPRG.Q SEQ ID NO: 713


 


MMP2, Alpha1
G.ETGFQGKTGPPGPPG.V SEQ ID NO: 714


 


MMP2, Alpha1
G.LRGFPGDRGLPGPV.G SEQ ID NO: 715


 


MMP2, Alpha1
G.LRGFPGDRGLPGPVG.A SEQ ID NO: 716


 


MMP2, Alpha1
G.KTGPIGPQGAPGKPGPDG.L SEQ ID NO: 717


 


MMP2, Alpha1
G.PPGRPGLPGADGLPGPPG.T SEQ ID NO: 718


 


MMP2, Alpha1
G.LKGNEGPPGPPGPAGSPGE.R SEQ ID NO: 719


 


MMP2, Alpha1
G.LRGFPGDRGLPGPVGALG.L SEQ ID NO: 720


 


MMP2, Alpha1
G.ERGHPGPPGPPGEQGLPG.L SEQ ID NO: 721


 


MMP2, Alpha1
I.GPPGEQGEKGDRGLPGPQG.S SEQ ID NO: 722


 


MMP2, Alpha1
G.EAGHPGPPGPPGPPGEVIQPLP.I SEQ ID NO: 723


 


MMP2, Alpha1
K.PGPKGNSGGDGPAGPPGERGPNGP.Q SEQ ID NO: 724


 


MMP2, Alpha1
G.EQGLPGSPGPDGPPGPMGPPGLPG.L SEQ ID NO: 725


 


MMP2, Alpha1
E.GPPGEKGGQGPPGPQGPIGYPGPRG.V SEQ ID NO: 726


 


MMP2, Alpha1
G.FPGPKGPPGPPGKDGLPGHPGQRG.E SEQ ID NO: 727


 


MMP2
L.PFRFGGGGDA SEQ ID NO: 728


 


MMP2 and 9
GSKGPMVSAQ.E SEQ ID NO: 729


 


MMP2 and 9
Q.ESQAQAILQQ SEQ ID NO: 730


 


MMP9, Alpha1
L.ALRGPAGPMG.L SEQ ID NO: 707


 


MMP9, Alpha1
G.AIGPPGEKGPLG.K SEQ ID NO: 731


 


MMP9, Alpha1
G.GPNGDPGPLGPPG.E SEQ ID NO: 732


 


MMP9, Alpha1
P.PGPPGEQGLPGL.A SEQ ID NO: 733


 


MMP9, Alpha1
G.LLGPKGPPGPPGPPG.V SEQ ID NO: 734


 


MMP9, Alpha1
G.IPGRPGPQGPPGPAG.E SEQ ID NO: 711


 


MMP9, Alpha1
G.QPGPSGADGEPGPRG.Q SEQ ID NO: 713


 


MMP9, Alpha1
G.QQGNPGAQGLPGPQG.A SEQ ID NO: 735


 


MMP9, Alpha1
G.KEGPPGEKGGQGPPG.P SEQ ID NO: 736


 


MMP9, Alpha1
G.ETGFQGKTGPPGPPG.V SEQ ID NO: 737


 


MMP9, Alpha1
G.EKGHPGLIGLIGPPG.E SEQ ID NO: 738


 


MMP9, Alpha1
G.LRGFPGDRGLPGPVG.A SEQ ID NO: 716


 


MMP9, Alpha1
G.KTGPIGPQGAPGKPGPDG.L SEQ ID NO: 739


 


MMP9, Alpha1
P.GPDGPPGPMGPPGLPGLK.G SEQ ID NO: 740


 


MMP9, Alpha1
G.ERGHPGPPGPPGEQGLPG.L SEQ ID NO: 721


 


MMP9, Alpha1
G.ERGPNGPQGPTGFPGPKGPPGPPG.K SEQ ID NO: 741


 


MMP9, Alpha1
L.IGLIGPPGEQGEKGDRGLPGPQGS.S SEQ ID NO: 742


 


MMP9, Alpha1
E.GPPGEKGGQGPPGPQGPIGYPGPRG.V SEQ ID NO: 726


 


MMP9, Alpha1
I.GPPGPPGLPGPPGPKGAKGSSGPTGPKGE.A SEQ ID NO: 743


 


MMP9, Alpha1
P.LGPPGEKGKLGVPGLPGYPGRQGPKGSI.G SEQ ID NO: 744


 


MMP9, Alpha1
Q.GPKGSIGFPGFPGANGEKGGRGTPGKPGPRG.Q SEQ ID NO: 745


 


MMP9, Alpha3
P.GPKGDPGPPGPIG.S SEQ ID NO: 746


 


MMP9, Alpha3
K.GDPGPPGPIGSLG.H SEQ ID NO: 683


 


MMP9, Alpha3
A.PGIPGEKGLPGL.Q SEQ ID NO: 747


 


MMP9, Alpha3
Q.GPPGPKGDPGPPGP.I SEQ ID NO: 748


 


MMP9, Alpha3
G.SLGHPGPPGVAGPLG.Q SEQ ID NO: 749


 


MMP9, Alpha3
G.KDGIPGPLGPLGPPG.A SEQ ID NO: 686


 


MMP9, Alpha3
G.VLGPQGKTGEVGPLG.E SEQ ID NO: 688


 


MMP9, Alpha3
G.ELGFQGQTGPPGPAG.V SEQ ID NO: 750


 


MMP9, Alpha3
G.EDGERGAEGPPGPTG.Q SEQ ID NO: 690


 


MMP9, Alpha3
G.LQGPPGFPGPKGPPG.H SEQ ID NO: 691


 


MMP9, Alpha3
G.EKGHIGLIGLIGPPG.E SEQ ID NO: 751


 


MMP9, Alpha3
G.QMGPPGPLGPSGLPGLK.G) SEQ ID NO: 694


 


MMP9, Alpha3
G.PVGEPGLLGAPGQMGPPG.P SEQ ID NO: 752


 


MMP9, Alpha3
G.LRGIPGPVGEPGLLGAPG.Q SEQ ID NO: 696


 


MMP9, Alpha3
G.LLGPRGSPGPTGRPGVTG.I SEQ ID NO: 697


 


MMP9, Alpha3
G.KDGIPGPLGPLGPPGAAGPSG.E SEQ ID NO: 700


 


MMP9, Alpha3
Q.GLPGLEGREGAKGELGPPGPLG.K SEQ ID NO: 701


 


MMP9, Alpha3
G.SRGERGPPGPTGKDGIPGPLGPLG.P SEQ ID NO: 753


 


MMP9, Alpha3
G.EKGKSGKTGQPGLEGERGPPGSRG.E SEQ ID NO: 754


 


MMP9, Alpha3
L.GPIGEKGKSGKTGQPGLEGERGPPGSRG.E SEQ ID NO: 702


 


MMP9, Alpha3
G.ANGSPGERGPLGPAGGIGLPGQSGSEGPVGPAG.K SEQ ID NO: 704


 


MMP9, Alpha3
G.LIGTPGEKGPPGNPGIPGLPGSDGPLGHPGHEGPTG.E SEQ ID NO: 705


 


MMP13, Alpha1
L.PGEPGPRG.L SEQ ID NO: 755


 


MMP13, Alpha1
A.LRGPAGPMG.L SEQ ID NO: 756


 


MMP13, Alpha1
G.LPGEPGPRG.L SEQ ID NO: 706


 


MMP13, Alpha1
L.ALRGPAGPMG.L SEQ ID NO: 707


 


MMP13, Alpha1
R.LALRGPAGPMG.L SEQ ID NO: 708


 


MMP13, Alpha1
G.LRGFPGDRGLPGPVG.A SEQ ID NO: 716


 


MMP13, Alpha1
Q.ESQAQAILQQARLA.L SEQ ID NO: 730


 


MMP13, Alpha1
P.GPDGPPGPMGPPGLPGLK.G SEQ ID NO: 740


 


MMP13, Alpha1
G.PQGAIGPPGEKGPLGKPGLPGMPGADGPPGHPG.K SEQ ID NO: 757


 


MMP13, Alpha1
A.GPMGLTGRPGPVGPPGSGGLKGEPGDVGPQGPRG.V SEQ ID NO: 758


 


MMP13, Alpha3
G.VLGPQGKTGEVGPLG.E SEQ ID NO: 688


 


MMP13, Alpha3
G.LRGIPGPVGEPGLLGAPG.Q SEQ ID NO: 696


 


MMP13, Alpha3
G.LRGIPGPVGEPGLLGAPGQMGPPGPLGPSG.L SEQ ID NO: 703


 


MMP13, Alpha3
G.LRGIPGPVGEPGLLGAPGQMGPPGPLGPSGLPG.L SEQ ID NO: 759









P is hydroxyproline, K indicates hydroxylysine, glycosylation, lipoxidation or cross linking.

The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type v collagen. Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:







TABLE 15a





N-terminal sequences of protease generated peptide


fragments of Collagen type V.







Collagen type V









GDPGPP SEQ ID NO: 373
LRGIPG SEQ ID NO: 760
IGPPGI SEQ ID NO: 761


 


LQGPPG SEQ ID NO: 62
IGSLGH SEQ ID NO: 762
IRGPPG SEQ ID NO: 763


 


ANGSPG SEQ ID NO: 764
LIGTPG SEQ ID NO: 765
LPGEPG SEQ ID NO: 766


 


IPGRPG SEQ ID NO: 767
GPDGPP SEQ ID NO: 768
QPGPSG SEQ ID NO: 769


 


LKGNEG SEQ ID NO: 770
ERGHPG SEQ ID NO: 771
GPPGEQ SEQ ID NO: 772


 


FPGPKG SEQ ID NO: 491
PFRFGG SEQ ID NO: 773
ESQAQA SEQ ID NO: 774


 


LLGPKG SEQ ID NO: 775
QQGNPG SEQ ID NO: 776
KEGPPG SEQ ID NO: 777


 


IGLIGP SEQ ID NO: 778
GPPGPP SEQ ID NO: 100
LGPPGE SEQ ID NO: 779


 


GPPGPK SEQ ID NO: 780
SLGHPG SEQ ID NO: 781
KDGIPG SEQ ID NO: 782


 


PVGEPG SEQ ID NO: 783
LRGIPG SEQ ID NO: 760
KDGIPG SEQ ID NO: 782


 


ANGSPG SEQ ID NO: 764
LIGTPG SEQ ID NO: 765
PGEPGP SEQ ID NO: 784


 


LLGAPG SEQ ID NO: 785
GLPGLE SEQ ID NO: 786
GIPGEK SEQ ID NO: 787


 


LALRGP SEQ ID NO: 788
LTGRPG SEQ ID NO: 789
LLGPKG SEQ ID NO: 775


 


LRGFPG SEQ ID NO: 790
KTGPIG SEQ ID NO: 791
PPGRPG SEQ ID NO: 792


 


PGPKGN SEQ ID NO: 527
EQGLPG SEQ ID NO: 793
GPPGEK SEQ ID NO: 640


 


AIGGPP SEQ ID NO: 794
GPNGDP SEQ ID NO: 795
PGPPGE SEQ ID NO: 796


 


LLGPRG SEQ ID NO: 797
GPDGPP SEQ ID NO: 768
ERGPNG SEQ ID NO: 798


 


GPKGDP SEQ ID NO: 799
GDPGPP SEQ ID NO: 373
PGIPGE SEQ ID NO: 800


 


LQGPPG SEQ ID NO: 62
EKGHIG SEQ ID NO: 801
QMGPPG SEQ ID NO: 802


 


SRGERG SEQ ID NO: 803
EKGKSG SEQ ID NO: 804
GPIGEK SEQ ID NO: 805


 


LPGEPG SEQ ID NO: 766
PQGAIG SEQ ID NO: 806
GPMGLT SEQ ID NO: 807


 


QMGPPG SEQ ID NO: 802
ETGFQG SEQ ID NO: 808
GSKGPM SEQ ID NO: 809


 


ALRGPA SEQ ID NO: 810
EAGHPG SEQ ID NO: 811
EKGHPG SEQ ID NO: 812


 


KDGIP SEQ ID NO: 813
VLGPQG SEQ ID NO: 814
EDGERG SEQ ID NO: 815


 


GPKGSI SEQ ID NO: 816
ELGFQG SEQ ID NO: 817
LRGPAG SEQ ID NO: 818









P is hydroxyproline, K indicates hydroxylysine, glycosylation, lipoxidation or cross linking.

or with any of the following sequences at the C-terminal of a peptide:








TABLE 16a





C-terminal sequences of protease generated peptide


fragments of Collagen type V.







Collagen type V









PIGSLG SEQ ID NO: 819
PVGEPG SEQ ID NO: 783
PGPPGE SEQ ID NO: 796


 


PPGPTG SEQ ID NO: 820
PKGPPG SEQ ID NO: 821
VAGPLG SEQ ID NO: 822


 


RPGVTG SEQ ID NO: 823
MMPFQF SEQ ID NO: 824
PGAAGP SEQ ID NO: 825


 


PVGPAG SEQ ID NO: 826
HEGPTG SEQ ID NO: 827
EPGPRG SEQ ID NO: 516


 


GPPGLP SEQ ID NO: 828
GQGPPG SEQ ID NO: 664
PPGPPG SEQ ID NO: 119


 


AGSPGE SEQ ID NO: 829
PVGALG SEQ ID NO: 830
EQGLPG SEQ ID NO: 793


 


YPGPRG SEQ ID NO: 831
HPGQRG SEQ ID NO: 832
GGGGDA SEQ ID NO: 833


 


LIGPPG SEQ ID NO: 834
GLPGLK SEQ ID NO: 835
PPGPPG SEQ ID NO: 119


 


PPGPIG SEQ ID NO: 174
KGLPGL SEQ ID NO: 836
PGPPGP SEQ ID NO: 458


 


QMGPPG SEQ ID NO: 802
LLGAPG SEQ ID NO: 785
RPGVTG SEQ ID NO: 823


 


QQARLA SEQ ID NO: 837
PPGHPG SEQ ID NO: 838
PQGPRG SEQ ID NO: 150


 


PLGPPG SEQ ID NO: 839
GEPGLL SEQ ID NO: 840
EVGPLG SEQ ID NO: 841


 


IMMPFQ SEQ ID NO: 842
GLPGLK SEQ ID NO: 835
PLGPSG SEQ ID NO: 843


 


AAGPSG SEQ ID NO: 844
PPGPLG SEQ ID NO: 845
PPGSRG SEQ ID NO: 846


 


PAGPMG SEQ ID NO: 847
PPGSGG SEQ ID NO: 848
PPGPPG SEQ ID NO: 119


 


GLPGPV SEQ ID NO: 849
LPGPVG SEQ ID NO: 850
KPGPDG SEQ ID NO: 851


 


LPGPQG SEQ ID NO: 852
VIQPLP SEQ ID NO: 853
RGPNGP SEQ ID NO: 854


 


PGPQGS SEQ ID NO: 855
EKGPLG SEQ ID NO: 856
PLGPPG SEQ ID NO: 839


 


GPPGAA SEQ ID NO: 857
TGPKGE SEQ ID NO: 858
GPKGSI SEQ ID NO: 816


 


PPGPAG SEQ ID NO: 52
PPGPAG SEQ ID NO: 52
PPGPTG SEQ ID NO: 820


 


QGLPGL SEQ ID NO: 859
PSGLPG SEQ ID NO: 860
LLGAPG SEQ ID NO: 785


 


PPGSRG SEQ ID NO: 846
LPGPPG SEQ ID NO: 72
PPGLPG SEQ ID NO: 861


 


PPGPLG SEQ ID NO: 845
KPGPRG SEQ ID NO: 862
PKGPPG SEQ ID NO: 821


 


PLGPLG SEQ ID NO: 863
PPGSRG SEQ ID NO: 846









P is hydroxyproline, K indicates hydroxylysine, glycosylation, lipoxidation or cross linking.

Collagen VI

We have determined that the enzymes listed in the following table cleave type vi collagen at least the following cleavage sites (marked “.” or in the ablsence of a ‘.’, at the end of the sequence):







TABLE 14b







Cleavage fragments of collagen type VI








Pro-



tease
Neoepitope





MMP2
G.YRGPEGPQGPPG.H SEQ ID NO: 864


 


MMP2
G.PIGPKGYRGDEGPP.G SEQ ID NO: 865


 


MMP2,
I.GIGIGNADIT.E SEQ ID NO: 866


(a3)


 


MMP2,
G.AQGPAGPAGPPG.L SEQ ID NO: 867


(a3)


 


MMP9
G.LIGEQGISGPRG.S SEQ ID NO: 868


 


MMP9
P.PGLIGEQGISGPR.G SEQ ID NO: 869


 


MMP9
E.PGEPGPKGGIGNRG.P SEQ ID NO: 870


 


MMP9
G.ISGPRGSGGAAGAPGERGRTGPLG.R SEQ ID NO: 871


 


MMP13
PGPAGPPGDPGLMG SEQ ID NO: 872


 


FAP-1
VAAKPAAVRPPAAAAAKPVATKPEVPRP SEQ ID NO: 873


 


FAP-1
GEPGLNGTTGPKGI SEQ ID NO: 874


 


FAP-1
IGPKGIPGEDGYRGYPG SEQ ID NO: 875


 


FAP-1
VAVVQHAPSESVDNASMPPVKVEFSL SEQ ID NO: 876


 


FAP-2
LGPMGVPGRD SEQ ID NO: 877


 


FAP-2
GEPGPPGEKGEAGDEGNPGPDGAPGERG SEQ ID NO: 878


 


FAP-2
RGPIGSIGPKGIPGEDGYRGYPGDEGGP SEQ ID NO: 879


 


FAP-2
PPPPQPARSAS SEQ ID NO: 880


 


FAP-2
FGPSAATPAPPG SEQ ID NO: 881


 


FAP-2
GPKGETGDLGPMGVPGRDGVPGGPGETGK SEQ ID NO: 882









The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neoepitope formed by cleavage of type v collagen. Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:







TABLE 15b





N-terminal sequences of protease generated peptide fragments


of Collagen type VI.







Collagen type VI









YRGPEG SEQ ID NO: 883
PIGPKG SEQ ID NO: 865
GIGIGN SEQ ID NO: 885


 


ISGPRG SEQ ID NO: 886
PGPAGP SEQ ID NO: 887
VAAKPA SEQ ID NO: 888


 


GEPGPP SEQ ID NO: 675
RGPIGS SEQ ID NO: 889
PPPPQP SEQ ID NO: 890


 


AQGPAG SEQ ID NO: 891
LIGEQG SEQ ID NO: 892
PGLIGE SEQ ID NO: 893


 


GEPGLN SEQ ID NO: 894
IGPKGI SEQ ID NO: 895
VAVVQH SEQ ID NO: 896


 


FGPSAA SEQ ID NO: 897
GPKGET SEQ ID NO: 898
PGEPGP SEQ ID NO: 784


 


LGPMGV SEQ ID NO: 899









or with any of the following sequences at the C-terminal of a peptide:








TABLE 16b





C-terminal sequences of protease generated peptide


fragments of Collagen type VI.







Collagen type VI









GDEGPP SEQ ID NO900
GNADIT SEQ ID NO901
PAGPPG SEQ ID NO133


 


DPGLMG SEQ ID NO902
PEVPRP SEQ ID NO903
TGPKGI SEQ ID NO904


 


GDEGGP SEQ ID NO905
PARSAS SEQ ID NO906
TPAPPG SEQ ID NO915


 


ISGPRG SEQ ID NO886
GISGPR SEQ ID NO907
GIGNRG SEQ ID NO908


 


YRGYPG SEQ ID NO909
KVEFSL SEQ ID NO910
GVPGRD SEQ ID NO911


 


PGETGK SEQ ID NO912
RTGPLG SEQ ID NO913
APGERG SEQ ID NO914









Proteoglycans

In another aspect of the invention, said peptide fragments are fragments of proteoglycans versican, lumican, perlecan, biglycan and decorin, which are all identified in fibrotic tissue. Several candidate proteases may be responsible for the digestion of proteoglycans in fibrotic lesions We have determined that the enzymes listed in table 17 generate lumican, versican, biglycan, perlecan and decorin resulting in at least following cleavage products:







TABLE 17







Cleavage fragments of biglycan, decorin,


versican, lumican, and perlecan.







Protease












Biglycan


MMP-3
SVPKEISPDTTLLDLQNNDISE SEQ ID NO: 916


 


MMP-3
KSVPKEISPDTTLLDLQNNDISE SEQ ID NO: 917


 


MMP-9
NSGFEPGAFDGLKLNYLRISEAK SEQ ID NO: 918


 


MMP-9
LKSVPKEISPDTTLLDLQNNDISE SEQ ID NO: 919


 


MMP-12
LRISEAKLTGIPKDLPET SEQ ID NO: 920


 


MMP-13
LKSVPKEISPDTTLLDLQNNDISE SEQ ID NO: 919


 


MMP-13
LTGIPKDLPETLNELHLDHNKIQAIE SEQ ID NO:



921


 


ADAMTS4
RISEAKLTGIPKDLPETLNE SEQ ID NO: 922


 


ADAMTS4
AIELEDLLRYSK SEQ ID NO: 923


 


ADAMTS4
AIELEDLLRY SEQ ID NO: 924


 


ADAMTS4
EAKLTGIPKDLPETLNE SEQ ID NO: 925


 


ADAMTS4
LKAVPKEISPDTTLLDLQNNDISE SEQ ID NO: 926


 


MMP-8
LLDLQNNDISELRKDD SEQ ID NO: 927


 


MMP-8
IELEDLLRYS SEQ ID NO: 928


 


CathepsinS
NSGFEPGAFDGLK SEQ ID NO: 929


 



Decorin


MMP-12
IVIELGTNPLK SEQ ID NO: 930


 


MMP-3
DEASGIGPEVPDDR SEQ ID NO: 931


 


MMP-3
LHLDGNKISRVDAAS SEQ ID NO: 932


 


MMP-3
VNNKISKVSPGAFTPL SEQ ID NO: 933


 


MMP-3
LILVNNKISKVSPGAFTPLVKLER SEQ ID NO: 934


 


MMP-9
SNPVQYWEIQPSTFR SEQ ID NO: 935


 


CathepsinK
SSGIENGAFQGMK SEQ ID NO: 884


 


CathepsinK
SSGIENGAFQGMKKLS SEQ ID NO: 946


 


ADAMTS1
KITEIKDGDFK SEQ ID NO: 936


 


ADAMTS1
GLPPSLTELHLDGNK SEQ ID NO: 937


 



Versican


Unknown
LLASDAGLYR SEQ ID NO: 938


 


Unknown
LATVGELQAAWR SEQ ID NO: 939


 


Unknown
ETTVLVAQNGNIK SEQ ID NO: 940


 



Lumican


Unknown
SLEDLQLTHNK SEQ ID NO: 941


 


Unknown
LKEDAVSAAFK SEQ ID NO: 942


 



Perlecan


Unknown
SIEYSPQLEDAGSR SEQ ID NO: 943


 


Unknown
LEGDTLIIPR SEQ ID NO: 944


 


ADAMTS4
VSEAVVEKLEPEYR SEQ ID NO: 945


 


ADAMTS4
EVSEAVVEKLEPEYR SEQ ID NO: 947


 


ADAMTS4
SIEYSPQLEDASAKEFR SEQ ID NO: 948









The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neo-epitope formed by cleavage of type versican, lumican, decorin, perlecan, and biglycan.
Suitable immunological binding partners may therefore be specifically reactive with any of the following at the N terminal of a peptide:







TABLE 18





N-terminal sequences of protease generated peptide fragments


of biglycan, decorin, lumican, versican, and perlecan.







Biglycan









SVPKEI SEQ ID NO: 949
GLKLNY SEQ ID NO: 950
RISEAK SEQ ID NO: 951


 


NSGFEP SEQ ID NO: 952
LKSVPK SEQ ID NO: 953
AIELED SEQ ID NO: 954


 


IELEDL SEQ ID NO: 957
QCSDLG SEQ ID NO: 955
EAKLTG SEQ ID NO: 956


 


LRISEA SEQ ID NO: 958
LTGIPK SEQ ID NO: 959
LKAVPK SEQ ID NO: 960


 


LLDLQN SEQ ID NO: 961


 







Decorin









IVIELG SEQ ID NO: 962
DEASGI SEQ ID NO: 963
VNNKIS SEQ ID NO: 964


 


NGLNQM SEQ ID NO: 965
LHLDGN SEQ ID NO: 966
LILVNN SEQ ID NO: 967


 


SSGIEN SEQ ID NO: 968
KITEIK SEQ ID NO: 969
GLPPSL SEQ ID NO: 970


 


SNPVQY SEQ ID NO: 971


 







Versican









LLASDA SEQ ID NO: 972
LATVGE SEQ ID NO: 973
ETTVLV SEQ ID NO: 974


 


ENQDAR SEQ ID NO: 975
NGFDQC SEQ ID NO: 976
SLTVVK SEQ ID NO: 977


 







Lumican


 









SLEDLQ SEQ ID NO: 978
LKEDAV SEQ ID NO: 979
HLQHNR SEQ ID NO: 980


 


LQHNRL SEQ ID NO: 985


 







Perlecan









SIEYSP SEQ ID NO: 981
LVNFTR SEQ ID NO: 982
VSEAVV SEQ ID NO: 983


 


EVSEAV SEQ ID NO: 984





 






or with any of the following sequences in table 19, at the C-terminal of a peptide:








TABLE 19





C-terminal sequences of protease generated peptide fragments of


biglycan, decorin, lumican, versican, and perlecan.







Biglycan









NNDISE SEQ ID NO: 986
YWEVQP SEQ ID NO: 987
EDLLRY SEQ ID NO: 988


 


RISEAK SEQ ID NO: 951
KIQAIE SEQ ID NO: 989
PETLNE SEQ ID NO: 990


 


LRKDDF SEQ ID NO: 991
LLRYSK SEQ ID NO: 992
ELRKDD SEQ ID NO: 993


 


KDLPET SEQ ID NO: 994
DLLRYS SEQ ID NO: 995
AFDGLK SEQ ID NO: 996


 


LNELHL SEQ ID NO: 997


 







Decorin









GTNPLK SEQ ID NO: 998
EVPDDR SEQ ID NO: 999
GAFTPL SEQ ID NO: 1000


 


SSGIEN SEQ ID NO: 968
RVDAAS SEQ ID NO: 1001
LVKLER SEQ ID NO: 1002


 


GMKKLS SEQ ID NO: 1003
KDGDFK SEQ ID NO: 1004
HLDGNK SEQ ID NO: 1005


 


QPSTFR SEQ ID NO: 1006
AFQGMK SEQ ID NO: 1007


 







Versican









CDVMYG SEQ ID NO: 1008
NGFDQC SEQ ID NO: 976
QNGINK SEQ ID NO: 1009


 


IGQDYK SEQ ID NO: 1010


 







Lumican









QLTHNK SEQ ID NO: 1011
VSAAFK SEQ ID NO: 1012
GLKSLE SEQ ID NO: 1013


 







Perlecan









EDAGSR SEQ ID NO: 1014
EFREVS SEQ ID NO: 1015
VAQQDS SEQ ID NO: 1016


 


SAKEFR SEQ ID NO: 1017
LEPEYR SEQ ID NO: 1018









CRP

Several candidate proteases may be responsible for the digestion of CRP in fibrotic tissue the literature reports many different proteases in fibrotic tissue. Most likely, this is the result of the large range of complicated processes eventually leading to fibrosis. However, in our assessment, early phases may consist of a range of MMPs, whereas later stages may rely more on cathepsin K degradation of the matrix, resulting in different neo-epitope profiles dependent on the levels of disease. We have through a range of in vitro cleavages of pure native proteins determined that the enzymes listed in the following tables of cleaved CRP at least following cleavage sites (marked * in Table 20, but at the ends of each sequence in Table 21):







TABLE 20







CRP fragments generated by specific proteases.








Protease/



Protein
Neo-epitope





CRP + CatK
K*ESDTSYVSLKAPLT*K SEQ ID NO: 1019


 


CRP + CatK
G*GNFEGSQSLVGDIG*N SEQ ID NO: 1020


 


CRP + MMP9
A*LKYEVQGEVFTKPQ*L SEQ ID NO: 1021


 


CRP + MMP9
G*IVEFWVDGKPRV*R SEQ ID NO: 1022


 


CRP + MMP1/
R*KAFVFPKE*S SEQ ID NO: 1023


MMP3


 


CRP + MMP3
K*YEVQGEVFTKPQLWP*- SEQ ID NO: 1024


 


CRP + MMP3
D*SFGGNFEGSQS*L SEQ ID NO: 1025


 


CRP + MMP3
D*FVLSPDEINT*I SEQ ID NO: 1026


 


CRP + MMP3
S*LKKGYTVGAEA*S SEQ ID NO: 1027


 


CRP + MMP3
A*FGQTDMSRKA*F SEQ ID NO: 1028


 


CRP + MMP3
S*LKKGYTVGAEAS*I SEQ ID NO: 1029


 


CRP + MMP3
G*EVFTKPQLWP*- SEQ ID NO: 1030


 


CRP + MMP3
S*IILGQEQDSFGGN.F SEQ ID NO: 1031


 


CRP + MMP3
K*YEVQGEVFTKPQ.L SEQ ID NO: 1032
















TABLE 21







CRP fragments generated by specific proteases.









Protease
Neoepitope
Aminoacid Nos*





MMP9
AFVFPK SEQ ID NO: 1033
026-031


 


MMP9
FGQTDMSR SEQ ID NO: 1034
017-024


 


MMP9
FGQTDMSRK SEQ ID NO: 1035
017-025


 


MMP9
FGQTDMSRKA SEQ ID NO: 1036
017-026


 


MMP9
FGQTDMSRKAF SEQ ID NO: 1037
017-027


 


MMP9
FGQTDMSRKAFVFPKE SEQ ID NO: 1038
017-032


 


MMP9
FGQTDMSRKAFVFPKESDTS SEQ ID NO: 1039
017-036


 


MMP9
FGQTDMSRKAFVFPKESDTSYV SEQ ID NO: 1040
017-038


 


MMP9
FGQTDMSRKAFVFPKESDTSYVS SEQ ID NO: 1041
017-039


 


MMP9
TDMSRKAFVFPKESDTSYV SEQ ID NO: 1042
020-038


 


MMP9
MSRKAFVFPKESDTS SEQ ID NO: 1043
022-036


 


MMP9
SRKAFVFPKESDTSYV SEQ ID NO: 1044
023-038


 


MMP9
RKAFVFPKE SEQ ID NO: 1045
024-032


 


MMP9
RKAFVFPKESDTSYV SEQ ID NO: 1046
024-038


 


MMP9
RKAFVFPKESDTSYVS SEQ ID NO: 1047
024-039


 


MMP9
KAFVFPKE SEQ ID NO: 1048
025-032


 


MMP9
KAFVFPKESD SEQ ID NO: 1049
025-034


 


MMP9
KAFVFPKESDT SEQ ID NO: 1050
025-035


 


MMP9
KAFVFPKESDTS SEQ ID NO: 1051
025-036


 


MMP9
KAFVFPKESDTSYV SEQ ID NO: 1052
025-038


 


MMP9
KAFVFPKESDTSYVS SEQ ID NO: 1053
025-039


 


MMP9
AFVFPKE SEQ ID NO: 1054
026-032


 


MMP9
AFVFPKESDT SEQ ID NO: 1055
026-035


 


MMP9
AFVFPKESDTSYV SEQ ID NO: 1056
026-038


 


MMP9
AFVFPKESDTSYVS SEQ ID NO: 1057
026-039


 


MMP9
AFVFPKESDTSYVSL SEQ ID NO: 1058
026-040


 


MMP9
FVFPK SEQ ID NO: 1059
027-031


 


MMP9
FVFPKE SEQ ID NO: 1060
027-032


 


MMP9
FVFPKESD SEQ ID NO: 1061
027-034


 


MMP9
FVFPKESDTS SEQ ID NO: 1062
027-036


 


MMP9
FVFPKESDTSY SEQ ID NO: 1063
027-037


 


MMP9
FVFPKESDTSYV SEQ ID NO: 1064
027-038


 


MMP9
FVFPKESDTSYVS SEQ ID NO: 1065
027-039


 


MMP9
FVFPKESDTSYVSL SEQ ID NO: 1066
027-040


 


MMP9
VFPKESDTS SEQ ID NO: 1067
028-036


 


MMP9
VFPKESDTSYV SEQ ID NO: 1068
028-038


 


MMP9
VFPKESDTSYVS SEQ ID NO: 1069
028-039


 


MMP9
VFPKESDTSYVSL SEQ ID NO: 1070
028-040


 


MMP9
FPKESDTSYVS SEQ ID NO: 1071
029-039


 


MMP9
KESDTSYVSLKAPLTKP SEQ ID NO: 1072
031-047


 


MMP9
SDTSYVSLKAPLTKP SEQ ID NO: 1073
033-047


 


MMP9
SLKAPLTKP SEQ ID NO: 1074
039-047


 


MMP9
SLKAPLTKPLK SEQ ID NO: 1075
039-049


 


MMP9
LKAPLTKPLK SEQ ID NO: 1076
040-049


 


MMP9
FYTELSSTRGYS SEQ ID NO: 1077
057-068


 


MMP9
LSSTRGYS SEQ ID NO: 1078
061-068


 


MMP9
SSTRGYS SEQ ID NO: 1079
062-068


 


MMP9
STRGYS SEQ ID NO: 1080
063-068


 


MMP9
IFSYATKRQ SEQ ID NO: 1081
069-077


 


MMP9
IFSYATKRQDNEILI SEQ ID NO: 1082
069-083


 


MMP9
SYATKRQDNEILI SEQ ID NO: 1083
071-083


 


MMP9
YATKRQDNEIL SEQ ID NO: 1084
072-082


 


MMP9
YATKRQDNEILI SEQ ID NO: 1085
072-083


 


MMP9
YATKRQDNEILIF SEQ ID NO: 1086
072-084


 


MMP9
TKRQDNEILI SEQ ID NO: 1087
074-083


 


MMP9
TKRQDNEILIF SEQ ID NO: 1088
074-084


 


MMP9
TKRQDNEILIFWSKDI SEQ ID NO: 1089
074-089


 


MMP9
KRQDNEILI SEQ ID NO: 1090
075-083


 


MMP9
KRQDNEILIF SEQ ID NO: 1091
075-084


 


MMP9
WSKDIGYS SEQ ID NO: 1092
085-092


 


MMP9
SKDIGYS SEQ ID NO: 1093
086-092


 


MMP9
IVEFWVDGKPRV SEQ ID NO: 1094
124-135


 


MMP9
EFWVDGKPR SEQ ID NO: 1095
126-134


 


MMP9
WVDGKPRV SEQ ID NO: 1096
128-135


 


MMP9
VDGKPRV SEQ ID NO: 1097
129-135


 


MMP9
SLKKGYTVGAE SEQ ID NO: 1098
138-148


 


MMP9
SLKKGYTVGAEA SEQ ID NO: 1099
138-149


 


MMP9
SLKKGYTVGAEAS SEQ ID NO: 1100
138-150


 


MMP9
LKKGYTV SEQ ID NO: 1101
139-145


 


MMP9
LKKGYTVG SEQ ID NO: 1102
139-146


 


MMP9
LKKGYTVGA SEQ ID NO: 1103
139-147


 


MMP9
LKKGYTVGAE SEQ ID NO: 1104
139-148


 


MMP9
LKKGYTVGAEA SEQ ID NO: 1105
139-149


 


MMP9
LKKGYTVGAEAS SEQ ID NO: 1106
139-150


 


MMP9
LKKGYTVGAEASI SEQ ID NO: 1107
139-151


 


MMP9
SIILGQEQDSFGGN SEQ ID NO: 1108
150-163


 


MMP9
SIILGQEQDSFGGNFEGSQ SEQ ID NO: 1109
150-168


 


MMP9
SIILGQEQDSFGGNFEGSQS SEQ ID NO: 1110
150-169


 


MMP9
IILGQEQDSFGGNFEGS SEQ ID NO: 1111
151-067


 


MMP9
IILGQEQDSFGGNFEGSQS SEQ ID NO: 1112
151-169


 


MMP9
ILGQEQDSFGGN SEQ ID NO: 1113
152-163


 


MMP9
ILGQEQDSFGGNFEGSQ SEQ ID NO: 1114
152-168


 


MMP9
ILGQEQDSFGGNFEGSQS SEQ ID NO: 1115
152-169


 


MMP9
LGQEQDSFGGNFEGSQ SEQ ID NO: 1116
153-168


 


MMP9
LGQEQDSFGGNFEGSQS SEQ ID NO: 1117
153-169


 


MMP9
GQEQDSFGGNFEGSQS SEQ ID NO: 1118
154-169


 


MMP9
SFGGNFEGSQS SEQ ID NO: 1119
159-169


 


MMP9
QSLVGDIGNVN SEQ ID NO: 1120
168-178


 


MMP9
INTIYLGGPFSPNV SEQ ID NO: 1121
189-202


 


MMP9
INTIYLGGPFSPNVLN SEQ ID NO: 1122
189-204


 


MMP9
IYLGGPFSPNVLN SEQ ID NO: 1123
192-204


 


MMP9
YLGGPFSPNVLN SEQ ID NO: 1124
193-204


 


MMP9
LGGPFSPN SEQ ID NO: 1125
194-201


 


MMP9
SPNVLNWRALKYEVQGEVFTKPQLWP SEQ ID NO: 1126
199-224


 


MMP9
LNWRA SEQ ID NO: 1127
203-207


 


MMP9
LNWRAL SEQ ID NO: 1128
203-208


 


MMP9
LNWRALK SEQ ID NO: 1129
203-209


 


MMP9
WRALKYE SEQ ID NO: 1130
205-211


 


MMP9
WRALKYEV SEQ ID NO: 1131
205-212


 


MMP9
WRALKYEVQGE SEQ ID NO: 1132
205-215


 


MMP9
ALKYEV SEQ ID NO: 1133
207-212


 


MMP9
LKYEVQ SEQ ID NO: 1134
208-213


 


MMP9
LKYEVQG SEQ ID NO: 1135
208-214


 


MMP9
LKYEVQGE SEQ ID NO: 1136
208-215


 


MMP9
LKYEVQGEVFTKP SEQ ID NO: 1137
208-220


 


MMP9
LKYEVQGEVFTKPQ SEQ ID NO: 1138
208-221


 


MMP9
LKYEVQGEVFTKPQLWP SEQ ID NO: 1139
208-224


 


MMP9
KYEVQGE SEQ ID NO: 1140
209-215


 


MMP9
KYEVQGEVFTKPQ SEQ ID NO: 1141
209-221


 


MMP9
KYEVQGEVFTKPQLWP SEQ ID NO: 1142
209-224


 


MMP9
YEVQGEVFTKP SEQ ID NO: 1143
210-220


 


MMP9
YEVQGEVFTKPQ SEQ ID NO: 1144
210-221


 


MMP9
YEVQGEVFTKPQLWP SEQ ID NO: 1145
210-224


 


MMP9
VQGEVFTKPQ SEQ ID NO: 1146
212-221


 


MMP9
VQGEVFTKPQLWP SEQ ID NO: 1147
212-224


 


MMP9
QGEVFTKPQ SEQ ID NO: 1148
213-221


 


MMP9
GEVFTKP SEQ ID NO: 1149
214-220


 


MMP9
GEVFTKPQ SEQ ID NO: 1150
214-221


 


MMP9
EVFTKPQ SEQ ID NO: 1151
215-221


 


MMP9
EVFTKPQLWP SEQ ID NO: 1152
215-224


 


MMP9
VFTKPQ SEQ ID NO: 1153
216-221


 


MMP9
FTKPQ SEQ ID NO: 1154
217-221


 


MMP9
FTKPQLWP SEQ ID NO: 1155
217-224


 


MMP9
TKPQLWP SEQ ID NO: 1156
218-224


 


MMP9
KPQLWP SEQ ID NO: 1157
219-224


 


MMP12
FGQTDMSRKA SEQ ID NO: 1036
017-026


 


MMP12
MSRKAFVFP SEQ ID NO: 1158
022-030


 


MMP12
MSRKAFVFPKE SEQ ID NO: 1159
022-032


 


MMP12
MSRKAFVFPKESD SEQ ID NO: 1160
022-034


 


MMP12
MSRKAFVFPKESDTS SEQ ID NO: 1043
022-036


 


MMP12
MSRKAFVFPKESDTSYVS SEQ ID NO: 1161
022-039


 


MMP12
SRKAFVFP SEQ ID NO: 1162
023-030


 


MMP12
SRKAFVFPKESD SEQ ID NO: 1163
023-034


 


MMP12
SRKAFVFPKESDTS SEQ ID NO: 1164
023-036


 


MMP12
RKAFVFP SEQ ID NO: 1165
024-030


 


MMP12
RKAFVFPKESD SEQ ID NO: 1166
024-034


 


MMP12
KAFVFP SEQ ID NO: 1167
025-030


 


MMP12
KAFVFPKE SEQ ID NO: 1048
025-032


 


MMP12
KAFVFPKESD SEQ ID NO: 1049
025-034


 


MMP12
AFVFPKE SEQ ID NO: 1054
026-032


 


MMP12
AFVFPKESDTS SEQ ID NO: 1168
026-036


 


MMP12
AFVFPKESDTSYVS SEQ ID NO: 1057
026-039


 


MMP12
FVFPKE SEQ ID NO: 1060
027-032


 


MMP12
FVFPKESD SEQ ID NO: 1061
027-034


 


MMP12
FVFPKESDTS SEQ ID NO: 1062
027-036


 


MMP12
FVFPKESDTSY SEQ ID NO: 1063
027-037


 


MMP12
FVFPKESDTSYVS SEQ ID NO: 1065
027-039


 


MMP12
VFPKESD SEQ ID NO: 1169
028-034


 


MMP12
KESDTSY SEQ ID NO: 1170
031-037


 


MMP12
KESDTSYVS SEQ ID NO: 1171
031-039


 


MMP12
VSLKAP SEQ ID NO: 1172
038-043


 


MMP12
LKAPLT SEQ ID NO: 1173
040-045


 


MMP12
LKAPLTKP SEQ ID NO: 1174
040-047


 


MMP12
YTELSSTRGYS SEQ ID NO: 1175
058-068


 


MMP12
LSSTRGYS SEQ ID NO: 1078
061-068


 


MMP12
STRGYS SEQ ID NO: 1080
063-068


 


MMP12
YATKRQDNE SEQ ID NO: 1176
072-080


 


MMP12
YATKRQDNEI SEQ ID NO: 1177
072-081


 


MMP12
YATKRQDNEIL SEQ ID NO: 1084
072-082


 


MMP12
TKRQDNEIL SEQ ID NO: 1178
074-082


 


MMP12
KRQDNEIL SEQ ID NO: 1179
075-082


 


MMP12
ILIFWSKD SEQ ID NO: 1180
081-088


 


MMP12
IFWSKD SEQ ID NO: 1181
083-088


 


MMP12
SKDIGYS SEQ ID NO: 1093
086-092


 


MMP12
WVDGKPRV SEQ ID NO: 1096
128-135


 


MMP12
WVDGKPRVR SEQ ID NO: 1182
128-136


 


MMP12
VRKSLKKGYTVGAEAS SEQ ID NO: 1183
135-150


 


MMP12
SLKKGYT SEQ ID NO: 1184
138-144


 


MMP12
SLKKGYTVG SEQ ID NO: 1185
138-146


 


MMP12
SLKKGYTVGA SEQ ID NO: 1186
138-147


 


MMP12
SLKKGYTVGAE SEQ ID NO: 1098
138-148


 


MMP12
SLKKGYTVGAEA SEQ ID NO: 1099
138-149


 


MMP12
SLKKGYTVGAEAS SEQ ID NO: 1100
138-150


 


MMP12
SLKKGYTVGAEASI SEQ ID NO: 1187
138-151


 


MMP12
LKKGYTV SEQ ID NO: 1101
139-145


 


MMP12
LKKGYTVG SEQ ID NO: 1102
139-146


 


MMP12
LKKGYTVGA SEQ ID NO: 1103
139-147


 


MMP12
LKKGYTVGAE SEQ ID NO: 1104
139-148


 


MMP12
LKKGYTVGAEA SEQ ID NO: 1105
139-149


 


MMP12
LKKGYTVGAEAS SEQ ID NO: 1106
139-150


 


MMP12
LKKGYTVGAEASI SEQ ID NO: 1107
139-151


 


MMP12
KKGYTVGAEAS SEQ ID NO: 1188
140-150


 


MMP12
KGYTVGAEAS SEQ ID NO: 1189
141-150


 


MMP12
KGYTVGAEASI SEQ ID NO: 1190
141-151


 


MMP12
SIILGQEQDSFGGN SEQ ID NO: 1108
150-163


 


MMP12
IILGQEQD SEQ ID NO: 1191
151-158


 


MMP12
IILGQEQDSFGGN SEQ ID NO: 1192
151-163


 


MMP12
IILGQEQDSFGGNFEGSQS SEQ ID NO: 1112
151-169


 


MMP12
ILGQEQDSFGGN SEQ ID NO: 1113
152-163


 


MMP12
LVGDIGNVNMWD SEQ ID NO: 1193
170-181


 


MMP12
INTIYLGGPFSPNVLN SEQ ID NO: 1122
189-204


 


MMP12
IYLGGPFSPN SEQ ID NO: 1194
192-201


 


MMP12
IYLGGPFSPNV SEQ ID NO: 1195
192-202


 


MMP12
IYLGGPFSPNVLN SEQ ID NO: 1123
192-204


 


MMP12
LGGPFSPNVLN SEQ ID NO: 1196
194-204


 


MMP12
WRALKYE SEQ ID NO: 1130
205-210


 


MMP12
YEVQGEVFTKP SEQ ID NO: 1143
210-220


 


MMP12
YEVQGEVFTKPQ SEQ ID NO: 1144
210-221


 


MMP12
YEVQGEVFTKPQLWP SEQ ID NO: 1145
210-224


 


MMP12
EVQGEVFTKP SEQ ID NO: 1197
211-220


 


MMP12
EVQGEVFTKPQLWP SEQ ID NO: 1198
211-224


 


MMP12
VQGEVFTKP SEQ ID NO: 1199
212-220


 


MMP12
VQGEVFTKPQ SEQ ID NO: 1146
212-221


 


MMP12
VQGEVFTKPQLWP SEQ ID NO: 1147
212-224


 


MMP12
GEVFTKPQLWP SEQ ID NO: 1200
214-224


 


MMP12
EVFTKP SEQ ID NO: 1201
215-220


 


MMP12
EVFTKPQLWP SEQ ID NO: 1152
215-224


 


MMP12
VFTKPQ SEQ ID NO: 1153
216-221


 


MMP12
VFTKPQL SEQ ID NO: 1202
216-222


 


MMP12
VFTKPQLWP SEQ ID NO: 1203
216-224


 


MMP12
FTKPQLWP SEQ ID NO: 1155
217-224


 


MMP12
TKPQLWP SEQ ID NO: 1156
218-224


 


MMP1
AFVFPK SEQ ID NO: 1033
006-031


 


MMP1
KAFVFPK SEQ ID NO: 1204
025-031


 


MMP1
VRKSLK SEQ ID NO: 1205
135-140


 


MMP1
YEVQGEVFTKPQLWP SEQ ID NO: 1145
210-224


 


MMP3
FGQTDMSRKA SEQ ID NO: 1036
017-026


 


MMP3
FGQTDMSRKAF SEQ ID NO: 1037
017-027


 


MMP3
MSRKAFVFPKESDTSYV SEQ ID NO: 1206
022-038


 


MMP3
MSRKAFVFPKESDTSYVS SEQ ID NO: 1161
022-039


 


MMP3
SRKAFVFPKESDTSYV SEQ ID NO: 1044
023-038


 


MMP3
SRKAFVFPKESDTSYVS SEQ ID NO: 1207
023-039


 


MMP3
RKAFVFPKESDTSYV SEQ ID NO: 1046
024-038


 


MMP3
RKAFVFPKESDTSYVS SEQ ID NO: 1047
024-039


 


MMP3
KAFVFPKE SEQ ID NO: 1048
025-032


 


MMP3
KAFVFPKESDTS SEQ ID NO: 1051
025-036


 


MMP3
KAFVFPKESDTSYVS SEQ ID NO: 1053
025-039


 


MMP3
KAFVFPKESDTSYVSL SEQ ID NO: 1208
025-040


 


MMP3
KAFVFPKESDTSYVSLK SEQ ID NO: 1209
025-041


 


MMP3
AFVFPKESDTSYVS SEQ ID NO: 1057
026-039


 


MMP3
AFVFPKESDTSYVSL SEQ ID NO: 1058
026-040


 


MMP3
AFVFPKESDTSYVSLKAP SEQ ID NO: 1210
026-043


 


MMP3
FVFPKESDTSYV SEQ ID NO: 1064
027-038


 


MMP3
FVFPKESDTSYVSLK SEQ ID NO: 1211
027-041


 


MMP3
VFPKESDTSYVSLK SEQ ID NO: 1212
028-041


 


MMP3
KESDTSYVSLKAP SEQ ID NO: 1213
031-043


 


MMP3
TKRQDNEILIFW SEQ ID NO: 1214
074-085


 


MMP3
IVEFWVDGKPRVRKS SEQ ID NO: 1215
124-138


 


MMP3
SLKKGYTVGAEA SEQ ID NO: 1099
138-149


 


MMP3
SLKKGYTVGAEAS SEQ ID NO: 1100
138-150


 


MMP3
LKKGYTVGAEA SEQ ID NO: 1105
139-149


 


MMP3
LKKGYTVGAEAS SEQ ID NO: 1106
139-150


 


MMP3
LKKGYTVGAEASI SEQ ID NO: 1107
139-151


 


MMP3
LKKGYTVGAEASII SEQ ID NO: 1216
139-152


 


MMP3
SIILGQEQDSFGGNFEGSQS SEQ ID NO: 1110
150-169


 


MMP3
IILGQEQDSFGGN SEQ ID NO: 1192
151-163


 


MMP3
IILGQEQDSFGGNFEGSQS SEQ ID NO: 1112
151-169


 


MMP3
ILGQEQDSFGGNFEGSQS SEQ ID NO: 1115
152-169


 


MMP3
LGQEQDSFGGNFEGSQS SEQ ID NO: 1117
153-169


 


MMP3
QEQDSFGGNFEGSQS SEQ ID NO: 1217
155-169


 


MMP3
SFGGNFEGSQS SEQ ID NO: 1119
159-169


 


MMP3
LVGDIGNVNMWD SEQ ID NO: 1193
170-181


 


MMP3
FVLSPDEINT SEQ ID NO: 1218
182-191


 


MMP3
YLGGPFSPNVLN SEQ ID NO: 1124
193-204


 


MMP3
LKYEVQGEVFTKPQ SEQ ID NO: 1138
208-221


 


MMP3
KYEVQGEVFTKPQ SEQ ID NO: 1141
209-221


 


MMP3
KYEVQGEVFTKPQLWP SEQ ID NO: 1142
209-224


 


MMP3
YEVQGEVFTKPQ SEQ ID NO: 1144
210-221


 


MMP3
YEVQGEVFTKPQLWP SEQ ID NO: 1145
210-224


 


MMP3
EVQGEVFTKPQLWP SEQ ID NO: 1198
211-224


 


MMP3
VQGEVFTKPQLWP SEQ ID NO: 1147
212-224


 


MMP3
GEVFTKPQLWP SEQ ID NO: 1200
214-224


 


MMP3
EVFTKPQLWP SEQ ID NO: 1152
215-224


 


MMP3
SKDIGYSFTVGGSEI SEQ ID NO: 1219
86-100


 


MMP8
FGQTDMSR SEQ ID NO: 1034
017-024


 


MMP8
FGQTDMSRK SEQ ID NO: 1035
017-025


 


MMP8
FGQTDMSRKA SEQ ID NO: 1036
017-026


 


MMP8
FGQTDMSRKAF SEQ ID NO: 1037
017-027


 


MMP8
FGQTDMSRKAFV SEQ ID NO: 1220
017-028


 


MMP8
FGQTDMSRKAFVFPKESDTSYV SEQ ID NO: 1040
017-038


 


MMP8
MSRKAFVFPKESDTSYV SEQ ID NO: 1206
022-038


 


MMP8
SRKAFVFPKESDTSYV SEQ ID NO: 1044
023-038


 


MMP8
RKAFVFPKESDTSYV SEQ ID NO: 1046
024-038


 


MMP8
KAFVFPKESDT SEQ ID NO: 1050
025-035


 


MMP8
KAFVFPKESDTS SEQ ID NO: 1051
025-036


 


MMP8
KAFVFPKESDTSYV SEQ ID NO: 1052
025-038


 


MMP8
KAFVFPKESDTSYVS SEQ ID NO: 1053
025-039


 


MMP8
AFVFPKESDTSYV SEQ ID NO: 1056
026-038


 


MMP8
FVFPKESDTSYV SEQ ID NO: 1064
027-038


 


MMP8
VFPKESDTSYV SEQ ID NO: 1068
028-038


 


MMP8
FPKESDTSYV SEQ ID NO: 1221
029-038


 


MMP8
SLKAPL SEQ ID NO: 1222
039-044


 


MMP8
SLKAPLTKP SEQ ID NO: 1074
039-047


 


MMP8
SLKAPLTKPLKA SEQ ID NO: 1223
039-050


 


MMP8
RGYSIFSYA SEQ ID NO: 1224
065-073


 


MMP8
FSYATKRQDNEILI SEQ ID NO: 1225
070-083


 


MMP8
SYATKRQDNEILI SEQ ID NO: 1083
071-083


 


MMP8
YATKRQDNEILI SEQ ID NO: 1085
072-083


 


MMP8
ATKRQDNEILI SEQ ID NO: 1226
073-083


 


MMP8
TKRQDNEILI SEQ ID NO: 1087
074-083


 


MMP8
TKRQDNEILIF SEQ ID NO: 1088
074-084


 


MMP8
FWSKDIGYS SEQ ID NO: 1227
084-092


 


MMP8
FWSKDIGYSFT SEQ ID NO: 1228
084-094


 


MMP8
FWSKDIGYSFTV SEQ ID NO: 1229
084-095


 


MMP8
WSKDIGYSFTV SEQ ID NO: 1230
085-095


 


MMP8
KSLKKGYTVGAEA SEQ ID NO: 1231
137-149


 


MMP8
SLKKGYTVGAEA SEQ ID NO: 1099
138-149


 


MMP8
LKKGYTV SEQ ID NO: 1101
139-145


 


MMP8
LKKGYTVGAEA SEQ ID NO: 1105
139-149


 


MMP8
LKKGYTVGAEAS SEQ ID NO: 1106
139-150


 


MMP8
KKGYTVGAEA SEQ ID NO: 1232
140-149


 


MMP8
GAEASIILGQE SEQ ID NO: 1233
146-156


 


MMP8
GAEASIILGQEQD SEQ ID NO: 1234
146-158


 


MMP8
SIILGQEQD SEQ ID NO: 1235
150-158


 


MMP8
SIILGQEQDSFGGNFEGSQ SEQ ID NO: 1109
150-168


 


MMP8
SIILGQEQDSFGGNFEGSQS SEQ ID NO: 1110
150-169


 


MMP8
IILGQEQDSFGGN SEQ ID NO: 1192
151-163


 


MMP8
IILGQEQDSFGGNFEGSQ SEQ ID NO: 1236
151-168


 


MMP8
IILGQEQDSFGGNFEGSQS SEQ ID NO: 1112
151-169


 


MMP8
ILGQEQDSFGGN SEQ ID NO: 1113
152-163


 


MMP8
ILGQEQDSFGGNFEGS SEQ ID NO: 1237
152-167


 


MMP8
ILGQEQDSFGGNFEGSQ SEQ ID NO: 1114
152-168


 


MMP8
ILGQEQDSFGGNFEGSQS SEQ ID NO: 1115
152-169


 


MMP8
LGQEQDSFGGN SEQ ID NO: 1238
153-163


 


MMP8
LGQEQDSFGGNFEGS SEQ ID NO: 1239
153-167


 


MMP8
LGQEQDSFGGNFEGSQ SEQ ID NO: 1116
153-168


 


MMP8
LGQEQDSFGGNFEGSQS SEQ ID NO: 1117
153-169


 


MMP8
LGQEQDSFGGNFEGSQSL SEQ ID NO: 1240
153-170


 


MMP8
LGQEQDSFGGNFEGSQSLV SEQ ID NO: 1241
153-171


 


MMP8
QDSFGGNFEGSQS SEQ ID NO: 1242
157-169


 


MMP8
SFGGNFEGSQ SEQ ID NO: 1243
159-168


 


MMP8
SFGGNFEGSQS SEQ ID NO: 1119
159-169


 


MMP8
SFGGNFEGSQSLV SEQ ID NO: 1244
159-171


 


MMP8
LVGDIGNVNMW SEQ ID NO: 1245
170-180


 


MMP8
INTIYLGGPFSPN SEQ ID NO: 1246
189-201


 


MMP8
TIYLGGPFSPN SEQ ID NO: 1247
191-201


 


MMP8
IYLGGPFSPN SEQ ID NO: 1194
192-201


 


MMP8
YLGGPFSPNV SEQ ID NO: 1248
193-202


 


MMP8
YLGGPFSPNVLN SEQ ID NO: 1124
193-204


 


MMP8
LGGPFSPNVLN SEQ ID NO: 1196
194-204


 


MMP8
VLNWRA SEQ ID NO: 1249
202-207


 


MMP8
VLNWRAL SEQ ID NO: 1250
202-208


 


MMP8
VLNWRALK SEQ ID NO: 1251
202-209


 


MMP8
LNWRAL SEQ ID NO: 1128
203-208


 


MMP8
LNWRALK SEQ ID NO: 1129
203-209


 


MMP8
LNWRALKYEV SEQ ID NO: 1252
203-212


 


MMP8
NWRAL SEQ ID NO: 1253
204-208


 


MMP8
NWRALKY SEQ ID NO: 1254
204-210


 


MMP8
NWRALKYEV SEQ ID NO: 1255
204-212


 


MMP8
NWRALKYEVQ SEQ ID NO: 1256
204-213


 


MMP8
WRALKYE SEQ ID NO: 1130
205-211


 


MMP8
WRALKYEVQ SEQ ID NO: 1257
205-213


 


MMP8
WRALKYEVQGE SEQ ID NO: 1132
205-215


 


MMP8
RALKYEV SEQ ID NO: 1258
206-212


 


MMP8
RALKYEVQ SEQ ID NO: 1259
206-213


 


MMP8
RALKYEVQGE SEQ ID NO: 1260
206-215


 


MMP8
ALKYEV SEQ ID NO: 1133
207-212


 


MMP8
ALKYEVQGEVFTKPQ SEQ ID NO: 1261
207-221


 


MMP8
LKYEVQGE SEQ ID NO: 1136
208-215


 


MMP8
LKYEVQGEVFTKPQ SEQ ID NO: 1138
208-221


 


MMP8
KYEVQGEVFTKPQ SEQ ID NO: 1141
209-221


 


MMP8
KYEVQGEVFTKPQLWP SEQ ID NO: 1142
209-224


 


MMP8
YEVQGEVFTKPQ SEQ ID NO: 1144
210-221


 


MMP8
YEVQGEVFTKPQLWP SEQ ID NO: 1145
210-224


 


MMP8
EVQGEVFTKPQ SEQ ID NO: 1262
211-221


 


MMP8
EVQGEVFTKPQLWP SEQ ID NO: 1198
211-224


 


MMP8
VQGEVFTKPQ SEQ ID NO: 1146
212-221


 


MMP8
VQGEVFTKPQLWP SEQ ID NO: 1147
212-224


 


MMP8
QGEVFTKPQ SEQ ID NO: 1148
213-221


 


MMP8
QGEVFTKPQL SEQ ID NO: 1263
213-222


 


MMP8
QGEVFTKPQLWP SEQ ID NO: 1264
213-224


 


MMP8
GEVFTKPQ SEQ ID NO: 1150
214-221


 


MMP8
GEVFTKPQLWP SEQ ID NO: 1200
214-224


 


MMP8
VFTKPQ SEQ ID NO: 1153
216-221


 


MMP8
VFTKPQLWP SEQ ID NO: 1203
216-224


 


MMP8
FTKPQLWP SEQ ID NO: 1155
217-224


 


MMP8
TKPQLWP SEQ ID NO: 1156
218-224


 


ADAMTS-1
ESDTSYVSLK SEQ ID NO: 1265
032-041


 


ADAMTS-1
QEQDSFGGNFEGSQ SEQ ID NO: 1266
155-168


 


ADAMTS-1
QEQDSFGGNFEGSQSLVG SEQ ID NO: 1267
155-172


 


ADAMTS-1
GNFEGSQSLVG SEQ ID NO: 1268
162-172


 


ADAMTS-1
YEVQGEVFT SEQ ID NO: 1269
210-218


 


ADAMTS-1
YEVQGEVFTKPQ SEQ ID NO: 1144
210-221


 


ADAMTS-1
GEVFTKPQ SEQ ID NO: 1150
214-221


 


ADAMTS-8
VFPKESDTSYVS SEQ ID NO: 1069
028-039


 


ADAMTS-8
QEQDSFGGNFEGSQSLVG SEQ ID NO: 1267
155-172


 


ADAMTS-8
EINTIYL SEQ ID NO: 1270
188-194


 


ADAMTS-8
KYEVQ SEQ ID NO: 1271
209-213


 


ADAMTS-8
KYEVQGE SEQ ID NO: 1140
209-215


 


Cat K
FGQTDMSR SEQ ID NO: 1034
017-024


 


Cat K
AFVFPK SEQ ID NO: 1033
026-031


 


Cat K
FVFPK SEQ ID NO: 1059
027-031


 


Cat K
ESDTSYVSLK SEQ ID NO: 1265
032-041


 


Cat K
ESDTSYVSLKAPLT SEQ ID NO: 1272
032-045


 


Cat K
SDTSYVSLK SEQ ID NO: 1273
033-041


 


Cat K
DTSYVSLK SEQ ID NO: 1274
034-041


 


Cat K
STRGYS SEQ ID NO: 1080
063-068


 


Cat K
IFWSKDIG SEQ ID NO: 1275
083-090


 


Cat K
KGYTVGAE SEQ ID NO: 1276
141-148


 


Cat K
AEASIILGQEQDSFG SEQ ID NO: 1277
147-161


 


Cat K
LGQEQDSFG SEQ ID NO: 1278
153-161


 


Cat K
LGQEQDSFGGNFE SEQ ID NO: 1279
153-165


 


Cat K
GQEQDSFG SEQ ID NO: 1280
154-161


 


Cat K
GQEQDSFGGNFE SEQ ID NO: 1281
154-165


 


Cat K
GQEQDSFGGNFEGSQ SEQ ID NO: 1282
154-168


 


Cat K
GQEQDSFGGNFEGSQS SEQ ID NO: 1118
154-169


 


Cat K
QEQDSFGGN SEQ ID NO: 1283
155-163


 


Cat K
QEQDSFGGNFE SEQ ID NO: 1284
155-165


 


Cat K
QEQDSFGGNFEG SEQ ID NO: 1285
155-166


 


Cat K
QEQDSFGGNFEGS SEQ ID NO: 1286
155-167


 


Cat K
QEQDSFGGNFEGSQ SEQ ID NO: 1266
155-168


 


Cat K
QEQDSFGGNFEGSQS SEQ ID NO: 1217
155-169


 


Cat K
GNFEGSQSLV SEQ ID NO: 1287
162-171


 


Cat K
GNFEGSQSLVG SEQ ID NO: 1268
162-172


 


Cat K
GNFEGSQSLVGDIG SEQ ID NO: 1288
162-175


 


Cat K
GSQSLVGDIG SEQ ID NO: 1289
166-175


 


Cat K
GSQSLVGDIGNVN SEQ ID NO: 1290
166-178


 


Cat K
DFVLSPDEIN SEQ ID NO: 1291
181-190


 


Cat K
FVLSPDEINT SEQ ID NO: 1218
182-191


 


Cat K
VLSPDEINT SEQ ID NO: 1291
183-191


 


Cat K
GPFSPNVLN SEQ ID NO: 1292
196-204


 


Cat K
SPNVLNWR SEQ ID NO: 1293
199-206


 


Cat K
KYEVQG SEQ ID NO: 1294
209-214


 


Cat K
YEVQGEVFT SEQ ID NO: 1269
210-218


 


Cat K
YEVQGEVFTKPQ SEQ ID NO: 1144
210-221


 


Cat K
VQGEVFTKPQ SEQ ID NO: 1146
212-221


 


Cat K
GEVFTKPQ SEQ ID NO: 1150
214-221


 


Cat K
EVFTKPQ SEQ ID NO: 1151
215-221


 


Cat S
FGQTDMSR SEQ ID NO: 1034
017-024


 


Cat S
AFVFPKESDTSYVS SEQ ID NO: 1057
026-039


 


Cat S
FVFPKESDTSYVS SEQ ID NO: 1065
027-039


 


Cat S
VFPKESDTSYVS SEQ ID NO: 1069
028-039


 


Cat S
FPKESDTSYVS SEQ ID NO: 1071
029-039


 


Cat S
ESDTSYVSLK SEQ ID NO: 1265
032-041


 


Cat S
TSWESASGIVE SEQ ID NO: 1295
116-126


 


Cat S
KGYTVG SEQ ID NO: 1296
141-146


 


Cat S
QEQDSFGGNFE SEQ ID NO: 1284
155-165


 


Cat S
QEQDSFGGNFEG SEQ ID NO: 1285
155-166


 


Cat S
QEQDSFGGNFEGSQ SEQ ID NO: 1266
155-168


 


Cat S
QEQDSFGGNFEGSQS SEQ ID NO: 1217
155-169


 


Cat S
QEQDSFGGNFEGSQSLV SEQ ID NO: 1297
155-171


 


Cat S
QEQDSFGGNFEGSQSLVG SEQ ID NO: 1267
155-172


 


Cat S
SFGGNFEGSQSLVG SEQ ID NO: 1298
159-172


 


Cat S
GNFEGSQSLVG SEQ ID NO: 1268
162-172


 


Cat S
GNFEGSQSLVGDIG SEQ ID NO: 1288
162-175


 


Cat S
SPDEINTIYL SEQ ID NO: 1299
185-194


 


Cat S
SPDEINTIYLG SEQ ID NO: 1300
185-195


 


Cat S
LGGPFSPNVLN SEQ ID NO: 1196
194-204


 


Cat S
GGPFSPNVLN SEQ ID NO: 1301
195-204


 


Cat S
GPFSPNVLN SEQ ID NO: 1292
196-204


 


Cat S
ALKYE SEQ ID NO: 1302
207-211


 


Cat S
ALKYEVQ SEQ ID NO: 1303
207-213


 


Cat S
YEVQGEVF SEQ ID NO: 1304
210-217


 


Cat S
YEVQGEVFT SEQ ID NO: 1269
210-218


 


Cat S
YEVQGEVFTKPQ SEQ ID NO: 1144
210-221


 


Cat S
YEVQGEVFTKPQLWP SEQ ID NO: 1145
210-224


 


Cat S
VQGEVFTKPQLWP SEQ ID NO: 1147
212-224


 


Cat S
GEVFTKPQ SEQ ID NO: 1150
214-221


 


Cat S
GEVFTKPQLWP SEQ ID NO: 1200
214-224


 


Cat S
EVFTKPQLWP SEQ ID NO: 1152
215-224


 


Cat S
TKPQLWP SEQ ID NO: 1156
218-224


 


Cat S
KPQLWP SEQ ID NO: 1157
219-224





*numbers in the sequence of CRP






Accordingly, in a method of the invention, said peptide fragments preferably comprise a neo-epitope formed by cleavage of CRP by a protease at a site marked by the sign * in any one of the above partial sequences of CRP in Table 20 or at either end of any partial sequence of CRP in Table 21.
The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neo-epitope formed by cleavage of CRP.
Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:







TABLE 22





N-terminal sequences of protease generated peptide


fragments of CRP.







CRP









AFVFPK SEQ ID NO: 1033
SFGGNF SEQ ID NO: 1305
FGQTDM SEQ ID NO: 1306


 


VSLKAP SEQ ID NO: 1172
KAFVFP SEQ ID NO: 1167
EVFTKP SEQ ID NO: 1201


 


TDMSRK SEQ ID NO: 1307
MSRKAF SEQ ID NO: 1308
SRKAFV SEQ ID NO: 1309


 


VFPKES SEQ ID NO: 1310
FPKESD SEQ ID NO: 1311
KESDTS SEQ ID NO: 1312


 


LSSTRG SEQ ID NO: 1313
SSTRGY SEQ ID NO: 1314
STRGYS SEQ ID NO: 1080


 


KRQDNE SEQ ID NO: 1315
WSKDIG SEQ ID NO: 1316
SKDIGY SEQ ID NO: 1317


 


SIILGQ SEQ ID NO: 1318
IILGQE SEQ ID NO: 1319
ILGQEQ SEQ ID NO: 1320


 


IYLGGP SEQ ID NO: 1321
YLGGPF SEQ ID NO: 1322
LGGPFS SEQ ID NO: 1323


 


ALKYEV SEQ ID NO: 1133
KYEVQG SEQ ID NO: 1294
VQGEVF SEQ ID NO: 1324


 


KPQLWP SEQ ID NO: 1157
YTELSS SEQ ID NO: 1325
ILIFWS SEQ ID NO: 1326


 


LVGDIG SEQ ID NO: 1327
QEQDSF SEQ ID NO: 1328
RGYSIF SEQ ID NO: 1329


 


GAEASI SEQ ID NO: 1330
QDSFGG SEQ ID NO: 1331
TIYLGG SEQ ID NO: 1332


 


EINTIY SEQ ID NO: 1333
DTSYVS SEQ ID NO: 1334
AEASII SEQ ID NO: 1335


 


TSWESA SEQ ID NO: 1336
SPDEIN SEQ ID NO: 1337
GGPFSP SEQ ID NO: 1338


 


YEVQGE SEQ ID NO: 1339
FVLSPD SEQ ID NO: 1340
LKKGYT SEQ ID NO: 1341


 


RKAFVF SEQ ID NO: 1342
IVEFWV SEQ ID NO: 1343
ESDTSY SEQ ID NO: 1344


 


TKPQLW SEQ ID NO: 1345
EVQGEV SEQ ID NO: 1346
FVFPK SEQ ID NO: 1059


 


SDTSYV SEQ ID NO: 1347
SLKAPL SEQ ID NO: 1222
LKAPLT SEQ ID NO: 1173


 


IFSYAT SEQ ID NO: 1348
SYATKR SEQ ID NO: 1349
YATKRQ SEQ ID NO: 1350


 


EFWVDG SEQ ID NO: 1351
WVDGKP SEQ ID NO: 1352
VDGKPR SEQ ID NO: 1353


 


LGQEQD SEQ ID NO: 1354
GQEQDS SEQ ID NO: 1355
QSLVGD SEQ ID NO: 1356


 


SPNVLN SEQ ID NO: 1357
LNWRA SEQ ID NO: 1127
LNWRAL SEQ ID NO: 1128


 


QGEVFT SEQ ID NO: 1358
GEVFTK SEQ ID NO: 1359
VFTKPQ SEQ ID NO: 1153


 


IFWSKD SEQ ID NO: 1181
VRKSLK SEQ ID NO: 1205
KKGYTV SEQ ID NO: 1360


 


FSYATK SEQ ID NO: 1361
ATKRQD SEQ ID NO: 1362
FWSKDI SEQ ID NO: 1363


 


VLNWRA SEQ ID NO: 1249
NWRAL SEQ ID NO: 1253
NWRALK SEQ ID NO: 1364


 


GSQSLV SEQ ID NO: 1365
DFVLSP SEQ ID NO: 1366
VLSPDE SEQ ID NO: 1367


 


LKYEVQ SEQ ID NO: 1134
TKRQDN SEQ ID NO: 1368
KGYTVG SEQ ID NO: 1296


 


GNFEGS SEQ ID NO: 1369
SLKKGY SEQ ID NO: 1370
KSLKKG SEQ ID NO: 1371


 


FVFPKE SEQ ID NO: 1060
INTIYL SEQ ID NO: 1372
RALKYE SEQ ID NO: 1373


 


FYTELS SEQ ID NO: 1374
WRALKY SEQ ID NO: 1375
GPFSPN SEQ ID NO: 1376









or with any of the following sequences at the C-terminal of a peptide:








TABLE 23





C-terminal sequences of protease generated peptide fragments of CRP.







CRP









AFVFPK SEQ ID NO: 1033
KPQLWP SEQ ID NO: 1157
PDEINT SEQ ID NO: 1377


 


SPDEIN SEQ ID NO: 1337
DSFGGN SEQ ID NO: 1378
VFTKPQ SEQ ID NO: 1153


 


KESDTS SEQ ID NO: 1312
SDTSYV SEQ ID NO: 1347
DTSYVS SEQ ID NO: 1334


 


LTKPLK SEQ ID NO: 1379
STRGYS SEQ ID NO: 1080
YATKRQ SEQ ID NO: 1350


 


KDIGYS SEQ ID NO: 1380
DGKPRV SEQ ID NO: 1381
VDGKPR SEQ ID NO: 1353


 


GAEASI SEQ ID NO: 1330
QGEVFT SEQ ID NO: 1358
NFEGSQ SEQ ID NO: 1382


 


SPNVLN SEQ ID NO: 1357
GPFSPN SEQ ID NO: 1376
RALKYE SEQ ID NO: 1373


 


ALKYEV SEQ ID NO: 1133
YEVQGE SEQ ID NO: 1339
LKYEVQ SEQ ID NO: 1134


 


KAFVFP SEQ ID NO: 1167
VSLKAP SEQ ID NO: 1172
LKAPLT SEQ IDNO 1173


 


LKKGYT SEQ ID NO: 1341
LGQEQD SEQ ID NO: 1354
NVNMWD SEQ ID NO: 1383


 


PRVRKS SEQ ID NO: 1384
TVGSEI SEQ ID NO: 1385
SRKAFV SEQ ID NO: 1309


 


YSFTV SEQ ID NO: 1386
IILGQE SEQ ID NO: 1319
EGSQSL SEQ ID NO: 1387


 


INTIYL SEQ ID NO: 1372
WSKDIG SEQ ID NO: 1316
EQDSFG SEQ ID NO: 1388


 


NVLNWR SEQ ID NO: 1389
ASGIVE SEQ ID NO: 1390
NTIYLG SEQ ID NO: 1391


 


VGAEAS SEQ ID NO: 1392
APLTKP SEQ ID NO: 1393
FVFPKE SEQ ID NO: 1060


 


QTDMSR SEQ ID NO: 1394
TDMSRK SEQ ID NO: 1307
MSRKAF SEQ ID NO: 1308


 


PKESDT SEQ ID NO: 1395
TSYVSL SEQ ID NO: 1396
ESDTSY SEQ ID NO: 1344


 


DNEILI SEQ ID NO: 1397
QDNEIL SEQ ID NO: 1398
NEILIF SEQ ID NO: 1399


 


YTVGAE SEQ ID NO: 1400
TVGAEA SEQ ID NO: 1401
PFSPNV SEQ ID NO: 1402


 


FEGSQS SEQ ID NO: 1403
GNFEGS SEQ ID NO: 1369
DIGNVN SEQ ID NO: 1404


 


LNWRA SEQ ID NO: 1127
LNWRAL SEQ ID NO: 1128
NWRALK SEQ ID NO: 1364


 


KYEVQG SEQ ID NO: 1294
EVFTKP SEQ ID NO: 1201
VFTKPQ SEQ ID NO: 1153


 


KRQDNE SEQ ID NO: 1315
RQDNEI SEQ ID NO: 1405
IFWSKD SEQ ID NO: 1181


 


FTKPQL SEQ ID NO: 1406
VRKSLK SEQ ID NO: 1205
SYVSLK SEQ ID NO: 1407


 


SLKAPL SEQ ID NO: 1222
TKPLKA SEQ ID NO: 1408
SIFSYA SEQ ID NO: 1409


 


GSQSLV SEQ ID NO: 1365
GNVNMW SEQ ID NO: 1410
SQSLVG SEQ ID NO: 1411


 


FGGNFE SEQ ID NO: 1412
GGNFEG SEQ ID NO: 1413
LVGDIG SEQ ID NO: 1327


 


VQGEVF SEQ ID NO: 1324
GKPRVR SEQ ID NO: 1414
FWSKDI 1363 SEQ ID NO:


 


DMSRKA SEQ ID NO: 1415
EILIFW SEQ ID NO: 1416
KKGYTV SEQ ID NO: 1360


 


FPKESD SEQ ID NO: 1311
IGYSFT SEQ ID NO: 1417









Elastin

Several candidate proteases may be responsible for the digestion of elastin in fibrotic tissue. We have through a range of in vitro cleavages of pure native proteins determined that the enzymes listed in the following table cleaved elastin at least at the cleavage sites at each end of the following sequences or at the cleavage sites marked ‘.’ or where no ‘.’ is shown, at the ends of the sequences:







TABLE 24







Elastin fragments generated by specific proteases.









Protease
Sequence between cleavage sites
Nos*





MMP9 + 12
GVPGAIPGGVPG SEQ ID NO: 1418
028-039


 


MMP9 + 12
AIPGGVPGGVFYPGAGLG SEQ ID NO: 1419
032-049


 


MMP9 + 12
AIPGGVPGGVFYPGAGLGA SEQ ID NO: 1420
032-050


 


MMP9 + 12
GVPGGVFYPGAGLGA SEQ ID NO: 1421
036-050


 


MMP9 + 12
GVPGGVFYPGAGLGALG SEQ ID NO: 1422
036-052


 


MMP9 + 12
VPGGVFYPGAGLGALGG SEQ ID NO: 1423
037-053


 


MMP9 + 12
GVFYPGAGLGALGGGALGPGG SEQ ID NO: 1424
040-060


 


MMP9 + 12
VFYPGAGLG SEQ ID NO: 1425
041-049


 


MMP9 + 12
VFYPGAGLGA SEQ ID NO: 1426
041-050


 


MMP9 + 12
VFYPGAGLGAL SEQ ID NO: 1427
041-051


 


MMP9 + 12
VFYPGAGLGALG SEQ ID NO: 1428
041-052


 


MMP9 + 12
VFYPGAGLGALGG SEQ ID NO: 1429
041-053


 


MMP9 + 12
VFYPGAGLGALGGG SEQ ID NO: 1430
041-054


 


MMP9 + 12
VFYPGAGLGALGGGAL SEQ ID NO: 1431
041-056


 


MMP9 + 12
VFYPGAGLGALGGGALG SEQ ID NO: 1432
041-057


 


MMP9 + 12
VFYPGAGLGALGGGALGPG SEQ ID NO: 1433
041-059


 


MMP9 + 12
VFYPGAGLGALGGGALGPGG SEQ ID NO: 1434
041-060


 


MMP9 + 12
VFYPGAGLGALGGGALGPGGKPLKPVPGG SEQ ID NO: 1435
041-069


 


MMP9 + 12
LGALGGGALGPGGKPLKPVPGG SEQ ID NO: 1436
048-069


 


MMP9 + 12
ALGGGALGPGGKPLKPVPGG SEQ ID NO: 1437
050-069


 


MMP9 + 12
LGGGALGPGGKPLKPVPG SEQ ID NO: 1438
051-068


 


MMP9 + 12
LGGGALGPGGKPLKPVPGG SEQ ID NO: 1439
051-069


 


MMP9 + 12
GGALGPGGKPLKPVPGG SEQ ID NO: 1440
053-069


 


MMP9 + 12
LGPGGKPLKPVPGG SEQ ID NO: 1441
056-069


 


MMP9 + 12
GPGGKPLKPVPGG SEQ ID NO: 1442
057-069


 


MMP9 + 12
PGGKPLKPVPGG SEQ ID NO: 1443
058-069


 


MMP9 + 12
GKPLKPVPGG SEQ ID NO: 1444
060-069


 


MMP9 + 12
PLKPVPGG SEQ ID NO: 1445
062-069


 


MMP9 + 12
LKPVPGG SEQ ID NO: SEQ ID NO: 1446
063-069


 


MMP9 + 12
GLAGAGLGAGLGAFP SEQ ID NO: 1447
069-083


 


MMP9 + 12
GLAGAGLGAGLGAFPA SEQ ID NO: 1448
069-084


 


MMP9 + 12
LAGAGLGAGLG SEQ ID NO: 1449
070-080


 


MMP9 + 12
LAGAGLGAGLGAFP SEQ ID NO: 1450
070-083


 


MMP9 + 12
LAGAGLGAGLGAFPA SEQ ID NO: 1451
070-084


 


MMP9 + 12
LAGAGLGAGLGAFPAVT SEQ ID NO: 1452
070-086


 


MMP9 + 12
LAGAGLGAGLGAFPAVTFPG SEQ ID NO: 1453
070-089


 


MMP9 + 12
LAGAGLGAGLGAFPAVTFPGA SEQ ID NO: 1454
070-090


 


MMP9 + 12
LAGAGLGAGLGAFPAVTFPGALVPGG SEQ ID NO: 1455
070-095


 


MMP9 + 12
LAGAGLGAGLGAFPAVTFPGALVPGGVA SEQ ID NO: 1456
070-097


 


MMP9 + 12
LAGAGLGAGLGAFPAVTFPGALVPGGVADAAAA
070-102



SEQ ID NO: 1457


 


MMP9 + 12
AGAGLGAGLGAFPAVTFPGALVPGG SEQ ID NO: 1458
071-095


 


MMP9 + 12
GAGLGAGLGAFPA SEQ ID NO: 1459
072-084


 


MMP9 + 12
GAGLGAGLGAFPAVTFPGA SEQ ID NO: 1460
072-090


 


MMP9 + 12
AGLGAGLGAFPA SEQ ID NO: 1461
073-084


 


MMP9 + 12
GLGAGLGAFPA SEQ ID NO: 1462
074-084


 


MMP9 + 12
LGAGLGAFPA SEQ ID NO: 1463
075-084


 


MMP9 + 12
LGAGLGAFPAVTFPGA SEQ ID NO: 1464
075-090


 


MMP9 + 12
LGAGLGAFPAVTFPGALVPGG SEQ ID NO: 1465
075-095


 


MMP9 + 12
LGAGLGAFPAVTFPGALVPGGVADAAAA SEQ ID NO: 1466
075-102


 


MMP9 + 12
AGLGAFPAVTFPG SEQ ID NO: 1467
077-089


 


MMP9 + 12
LGAFPAVTFPGA SEQ ID NO: 1468
079-090


 


MMP9 + 12
LGAFPAVTFPGALVPGGVA SEQ ID NO: 1469
079-097


 


MMP9 + 12
LGAFPAVTFPGALVPGGVADAAAA SEQ ID NO: 1470
079-102


 


MMP9 + 12
AFPAVTFPGALVPGG SEQ ID NO: 1471
081-095


 


MMP9 + 12
AVTFPGALVPGG SEQ ID NO: 1472
084-095


 


MMP9 + 12
AVTFPGALVPGGVADAAAA SEQ ID NO: 1473
084-102


 


MMP9 + 12
VTFPGALVPGG SEQ ID NO: 1474
085-095


 


MMP9 + 12
VTFPGALVPGGVADAAAA SEQ ID NO: 1475
085-102


 


MMP9 + 12
LVPGGVADAAAA SEQ ID NO: 1476
091-102


 


MMP9 + 12
LVPGGVADAAAAYK SEQ ID NO: 1477
091-104


 


MMP9 + 12
VADAAAAYK SEQ ID NO: 1478
096-104


 


MMP9 + 12
KAAKAGA SEQ ID NO: 1479
104-110


 


MMP9 + 12
LGVSAGAVVPQPGA SEQ ID NO: 1480
121-134


 


MMP9 + 12
VPGVGLPGVYPGGVLPGAR SEQ ID NO: 1481
141-159


 


MMP9 + 12
PGVGLPGVYPGGVLPGAR SEQ ID NO: 1482
142-159


 


MMP9 + 12
GLPGVYPGGVLPGAR SEQ ID NO: 1483
145-159


 


MMP9 + 12
PGVYPGGVLPGAR SEQ ID NO: 1484
147-159


 


MMP9 + 12
ARFPGVG SEQ ID NO: 1485
158-164


 


MMP9 + 12
ARFPGVGVLPG SEQ ID NO: 1486
158-168


 


MMP9 + 12
RFPGVGVLPGVPTGAG SEQ ID NO: 1487
159-174


 


MMP9 + 12
FPGVGVLPGVPTG SEQ ID NO: 1488
160-172


 


MMP9 + 12
FPGVGVLPGVPTGA SEQ ID NO: 1489
160-173


 


MMP9 + 12
FPGVGVLPGVPTGAGV SEQ ID NO: 1490
160-175


 


MMP9 + 12
FPGVGVLPGVPTGAGVKPK SEQ ID NO: 1491
160-178


 


MMP9 + 12
KPKAPGV SEQ ID NO: 1492
176-182


 


MMP9 + 12
PKAPGV SEQ ID NO: 1493
177-182


 


MMP9 + 12
GAFAGIPGVGPFG SEQ ID NO: 1494
184-196


 


MMP9 + 12
VGPFGGPQPGVPLGYP SEQ ID NO: 1495
192-207


 


MMP9 + 12
GPQPGVPLGYP SEQ ID NO: 1496
197-207


 


MMP9 + 12
PQPGVPLGYP SEQ ID NO: 1497
198-207


 


MMP9 + 12
PGVPLGYP SEQ ID NO: 1498
200-207


 


MMP9 + 12
GYPIKAPK SEQ ID NO: 1499
205-212


 


MMP9 + 12
PKLPGGY SEQ ID NO: 1500
211-217


 


MMP9 + 12
YTTGKLPYGYGPG SEQ ID NO: 1501
221-233


 


MMP9 + 12
YTTGKLPYGYGPGGVAGAAGK SEQ ID NO: 1502
221-241


 


MMP9 + 12
TTGKLPYGYG SEQ ID NO: 1503
222-231


 


MMP9 + 12
TTGKLPYGYGPGGVAGAAGK SEQ ID NO: 1504
222-241


 


MMP9 + 12
LPYGYGPGGVAGAAGK SEQ ID NO: 1505
226-241


 


MMP9 + 12
GYGPGGVAGAAGK SEQ ID NO: 1506
229-241


 


MMP9 + 12
YGPGGVAGAAGK SEQ ID NO: 1507
230-241


 


MMP9 + 12
AGYPTGTGVGPQAAAAAAAK SEQ ID NO: 1508
242-261


 


MMP9 + 12
TGVGPQAAAAAAAK SEQ ID NO: 1509
248-261


 


MMP9 + 12
PQAAAAAAAK SEQ ID NO: 1510
252-261


 


MMP9 + 12
FGAGAAGVLPGVGGAGVPGVPGAIPGIGG SEQ ID NO: 1511
266-294


 


MMP9 + 12
FGAGAAGVLPGVGGAGVPGVPGAIPGIGGIAGVGTPAA
266-303



SEQ ID NO: 1512


 


MMP9 + 12
GVLPGVGGAGVPGVPG SEQ ID NO: 1513
272-287


 


MMP9 + 12
VLPGVGGAGVPGVPGAIPGIGG SEQ ID NO: 1514
273-294


 


MMP9 + 12
VLPGVGGAGVPGVPGAIPGIGGIAGVGTPA SEQ ID NO: 1515
273-302


 


MMP9 + 12
VLPGVGGAGVPGVPGAIPGIGGIAGVGTPAA SEQ ID NO: 1516
273-303


 


MMP9 + 12
GAGVPGVPGAIPG SEQ ID NO: 1517
279-291


 


MMP9 + 12
GAGVPGVPGAIPGIGGIAGVG SEQ ID NO: 1518
279-299


 


MMP9 + 12
AGVPGVPGAIPGIG SEQ ID NO: 1519
280-293


 


MMP9 + 12
AGVPGVPGAIPGIGG SEQ ID NO: 1520
280-294


 


MMP9 + 12
AGVPGVPGAIPGIGGIAG SEQ ID NO: 1521
280-297


 


MMP9 + 12
AGVPGVPGAIPGIGGIAGVGTPA SEQ ID NO: 1522
280-302


 


MMP9 + 12
GVPGVPGAIPGIGG SEQ ID NO: 1523
281-294


 


MMP9 + 12
GVPGVPGAIPGIGGIA SEQ ID NO: 1524
281-296


 


MMP9 + 12
GVPGVPGAIPGIGGIAGVG SEQ ID NO: 1525
281-299


 


MMP9 + 12
VPGVPGAIPGIGG SEQ ID NO: 1526
282-294


 


MMP9 + 12
GVPGAIPGIGGIAGVGTPA SEQ ID NO: 1527
284-302


 


MMP9 + 12
VPGAIPGIGGIAGVG SEQ ID NO: 1528
285-299


 


MMP9 + 12
VPGAIPGIGGIAGVGTPA SEQ ID NO: 1529
285-302


 


MMP9 + 12
VPGAIPGIGGIAGVGTPAAA SEQ ID NO: 1530
285-304


 


MMP9 + 12
VPGAIPGIGGIAGVGTPAAAAAAAAAAK SEQ ID NO: 1531
285-312


 


MMP9 + 12
AIPGIGGIAGVG SEQ ID NO: 1532
288-299


 


MMP9 + 12
AIPGIGGIAGVGTPA SEQ ID NO: 1533
288-302


 


MMP9 + 12
AIPGIGGIAGVGTPAA SEQ ID NO: 1534
288-303


 


MMP9 + 12
AIPGIGGIAGVGTPAAA SEQ ID NO: 1535
288-304


 


MMP9 + 12
AIPGIGGIAGVGTPAAAAAA SEQ ID NO: 1536
288-307


 


MMP9 + 12
AIPGIGGIAGVGTPAAAAAAAAAAK SEQ ID NO: 1537
288-312


 


MMP9 + 12
IPGIGGIAGVGTPAAA SEQ ID NO: 1538
289-304


 


MMP9 + 12
IGGIAGVGTPAAAA SEQ ID NO: 1539
292-305


 


MMP9 + 12
GIAGVGTPAAAA SEQ ID NO: 1540
294-305


 


MMP9 + 12
GIAGVGTPAAAAAAAA SEQ ID NO: 1541
294-309


 


MMP9 + 12
GIAGVGTPAAAAAAAAAAK SEQ ID NO: 1542
294-312


 


MMP9 + 12
IAGVGTPAAAAAAAA SEQ ID NO: 1543
295-309


 


MMP9 + 12
IAGVGTPAAAAAAAAA SEQ ID NO: 1544
295-310


 


MMP9 + 12
IAGVGTPAAAAAAAAAAK SEQ ID NO: 1545
295-312


 


MMP9 + 12
TPAAAAAAAAAAK SEQ ID NO: 1546
300-312


 


MMP9 + 12
PAAAAAAAAAAK SEQ ID NO: 1547
301-312


 


MMP9 + 12
AAAAAAAAAAK SEQ ID NO: 1548
302-312


 


MMP9 + 12
AAAAAAAAAK SEQ ID NO: 1549
303-312


 


MMP9 + 12
AAAAAAAAK SEQ ID NO: 1550
304-312


 


MMP9 + 12
AAAAAAAAKA SEQ ID NO: 1551
304-313


 


MMP9 + 12
AAAAAAAK SEQ ID NO: 1552
305-312


 


MMP9 + 12
LVPGGPGFGPGVVGVPGA SEQ ID NO: 1553
322-339


 


MMP9 + 12
GPGFGPGVVGVPG SEQ ID NO: 1554
326-338


 


MMP9 + 12
GPGFGPGVVGVPGAGVPGVG SEQ ID NO: 1555
326-345


 


MMP9 + 12
GPGFGPGVVGVPGAGVPGVGVPGAGIPVVPG SEQ ID NO: 1556
326-356


 


MMP9 + 12
PGFGPGVVGVPG SEQ ID NO: 1557
327-338


 


MMP9 + 12
PGFGPGVVGVPGA SEQ ID NO: 1558
327-339


 


MMP9 + 12
PGFGPGVVGVPGAG SEQ ID NO: 1559
327-340


 


MMP9 + 12
PGVVGVPGAGVPG SEQ ID NO: 1560
331-343


 


MMP9 + 12
PGVVGVPGAGVPGVGVPG SEQ ID NO: 1561
331-348


 


MMP9 + 12
PGVVGVPGAGVPGVGVPGAGIPVVPGA SEQ ID NO: 1562
331-357


 


MMP9 + 12
VVGVPGAGVPGVGVPGA SEQ ID NO: 1563
333-349


 


MMP9 + 12
VGVPGAGVPGVGVPGAGIPVVPGAGIPGAAVPGVVSPEA
334-372



SEQ ID NO: 1564


 


MMP9 + 12
AGVPGVGVPGAGIPVVPG SEQ ID NO: 1565
339-356


 


MMP9 + 12
GVPGVGVPGAGIPVVPG SEQ ID NO: 1566
340-356


 


MMP9 + 12
GVPGVGVPGAGIPVVPGA SEQ ID NO: 1567
340-357


 


MMP9 + 12
VPGVGVPGAGIPVVPG SEQ ID NO: 1568
341-356


 


MMP9 + 12
VGVPGAGIPVVPG SEQ ID NO: 1569
344-356


 


MMP9 + 12
VGVPGAGIPVVPGAGIPG SEQ ID NO: 1570
344-361


 


MMP9 + 12
VPGAGIPVVPG SEQ ID NO: 1571
346-356


 


MMP9 + 12
AGIPVVPGAGIPG SEQ ID NO: 1572
349-361


 


MMP9 + 12
AGIPVVPGAGIPGAAVPGVVSPEAAAK SEQ ID NO: 1573
349-375


 


MMP9 + 12
GIPVVPGAGIPG SEQ ID NO: 1574
350-361


 


MMP9 + 12
IPGAAVPGVVSPEAAAK SEQ ID NO: 1575
359-375


 


MMP9 + 12
GAAVPGVVSPEAAAK SEQ ID NO: 1576
361-375


 


MMP9 + 12
AVPGVVSPEAAAK SEQ ID NO: 1577
363-375


 


MMP9 + 12
VPGVVSPEAAAK SEQ ID NO: 1578
364-375


 


MMP9 + 12
YGARPGVG SEQ ID NO: 1579
383-390


 


MMP9 + 12
YGARPGVGVG SEQ ID NO: 1580
383-392


 


MMP9 + 12
YGARPGVGVGGIPT SEQ ID NO: 1581
383-396


 


MMP9 + 12
YGARPGVGVGGIPTY SEQ ID NO: 1582
383-397


 


MMP9 + 12
YGARPGVGVGGIPTYG SEQ ID NO: 1583
383-398


 


MMP9 + 12
YGARPGVGVGGIPTYGVG SEQ ID NO: 1584
383-400


 


MMP9 + 12
YGARPGVGVGGIPTYGVGA SEQ ID NO: 1585
383-401


 


MMP9 + 12
YGARPGVGVGGIPTYGVGAG SEQ ID NO: 1586
383-402


 


MMP9 + 12
GARPGVGV SEQ ID NO: 1587
384-391


 


MMP9 + 12
GARPGVGVGG SEQ ID NO: 1588
384-393


 


MMP9 + 12
GARPGVGVGGIP SEQ ID NO: 1589
384-395


 


MMP9 + 12
GARPGVGVGGIPTY SEQ ID NO: 1590
384-397


 


MMP9 + 12
GARPGVGVGGIPTYGV SEQ ID NO: 1591
384-399


 


MMP9 + 12
GARPGVGVGGIPTYGVG SEQ ID NO: 1592
384-400


 


MMP9 + 12
GARPGVGVGGIPTYGVGAGGF SEQ ID NO: 1593
384-404


 


MMP9 + 12
GARPGVGVGGIPTYGVGAGGFPGF SEQ ID NO: 1594
384-407


 


MMP9 + 12
GARPGVGVGGIPTYGVGAGGFPGFG SEQ ID NO: 1595
384-408


 


MMP9 + 12
GARPGVGVGGIPTYGVGAGGFPGFGVGVG SEQ ID NO: 1596
384-412


 


MMP9 + 12
ARPGVGVGG SEQ ID NO: 1597
385-393


 


MMP9 + 12
ARPGVGVGGIP SEQ ID NO: 1598
385-395


 


MMP9 + 12
ARPGVGVGGIPTY SEQ ID NO: 1599
385-397


 


MMP9 + 12
ARPGVGVGGIPTYGVGA SEQ ID NO: 1600
385-401


 


MMP9 + 12
ARPGVGVGGIPTYGVGAGG SEQ ID NO: 1601
385-403


 


MMP9 + 12
ARPGVGVGGIPTYGVGAGGFPG SEQ ID NO: 1602
385-406


 


MMP9 + 12
ARPGVGVGGIPTYGVGAGGFPGF SEQ ID NO: 1603
385-407


 


MMP9 + 12
RPGVGVG SEQ ID NO: 1604
386-392


 


MMP9 + 12
RPGVGVGG SEQ ID NO: 1605
386-393


 


MMP9 + 12
PGVGVGGIPTY SEQ ID NO: 1606
387-397


 


MMP9 + 12
PGVGVGGIPTYG SEQ ID NO: 1607
387-398


 


MMP9 + 12
PGVGVGGIPTYGVGAG SEQ ID NO: 1608
387-412


 


MMP9 + 12
VGGIPTYGVGAG SEQ ID NO: 1609
391-402


 


MMP9 + 12
GVGAGGFPGFGVGVGGIPGVA SEQ ID NO: 1610
398-418


 


MMP9 + 12
VGAGGFPGFGVGVG SEQ ID NO: 1611
399-412


 


MMP9 + 12
VGVGGIPGVAGVPSVGGVPGVGGVPGVGISPEA SEQ ID NO: 1612
409-441


 


MMP9 + 12
VAGVPSVGGVPGVGGVPG SEQ ID NO: 1613
417-434


 


MMP9 + 12
VAGVPSVGGVPGVGGVPGVGISPEA SEQ ID NO: 1614
417-441


 


MMP9 + 12
SVGGVPGVGGVPGVGISPEA SEQ ID NO: 1615
422-441


 


MMP9 + 12
VGGVPGVGGVPGVGISPEA SEQ ID NO: 1616
423-441


 


MMP9 + 12
GVPGVGGVPGVGIS SEQ ID NO: 1617
425-438


 


MMP9 + 12
GVPGVGGVPGVGISPEA SEQ ID NO: 1618
425-441


 


MMP9 + 12
GVPGVGGVPGVGISPEAQA SEQ ID NO: 1619
425-443


 


MMP9 + 12
GVPGVGISPEAQAAAAAK SEQ ID NO: 1620
431-448


 


MMP9 + 12
GVGTPAAAAAK SEQ ID NO: 1621
482-492


 


MMP9 + 12
TPAAAAAK SEQ ID NO: 1622
485-492


 


MMP9 + 12
FGLVPGVGVAPGVG SEQ ID NO: 1623
500-513


 


MMP9 + 12
FGLVPGVGVAPGVGVAPG SEQ ID NO: 1624
500-517


 


MMP9 + 12
FGLVPGVGVAPGVGVAPGVGVAPG SEQ ID NO: 1625
500-523


 


MMP9 + 12
FGLVPGVGVAPGVGVAPGVGVAPGVG SEQ ID NO: 1626
500-525


 


MMP9 + 12
FGLVPGVGVAPGVGVAPGVGVAPGVGLAPG SEQ ID NO: 1627
500-529


 


MMP9 + 12
FGLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPG
500-535



SEQ ID NO: 1628


 


MMP9 + 12
FGLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPGV
500-536



SEQ ID NO: 1629


 


MMP9 + 12
FGLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPGVGVAPG
500-541



SEQ ID NO: 1630


 


MMP9 + 12
GLVPGVGVAPG SEQ ID NO: 1631
501-511


 


MMP9 + 12
GLVPGVGVAPGV SEQ ID NO: 1632
501-512


 


MMP9 + 12
GLVPGVGVAPGVGVA SEQ ID NO: 1633
501-515


 


MMP9 + 12
GLVPGVGVAPGVGVAP SEQ ID NO: 1634
501-516


 


MMP9 + 12
GLVPGVGVAPGVGVAPG SEQ ID NO: 1635
501-517


 


MMP9 + 12
GLVPGVGVAPGVGVAPGVG SEQ ID NO: 1636
501-519


 


MMP9 + 12
GLVPGVGVAPGVGVAPGVGVAPG SEQ ID NO: 1637
501-523


 


MMP9 + 12
GLVPGVGVAPGVGVAPGVGVAPGVGL SEQ ID NO: 1638
501-524


 


MMP9 + 12
GLVPGVGVAPGVGVAPGVGVAPGVGLA SEQ ID NO: 1639
501-525


 


MMP9 + 12
GLVPGVGVAPGVGVAPGVGVAPGVGLAPG SEQ ID NO: 1640
501-527


 


MMP9 + 12
GLVPGVGVAPGVGVAPGVGVAPGVGLAPGVG SEQ ID NO: 1641
501-529


 


MMP9 + 12
GLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVA
501-531



SEQ ID NO: 1642


 


MMP9 + 12
GLVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPG
501-533



SEQ ID NO: 1643


 


MMP9 + 12
LVPGVGVAPGVG SEQ ID NO: 1644
502-513


 


MMP9 + 12
LVPGVGVAPGVGVAPG SEQ ID NO: 1645
502-517


 


MMP9 + 12
LVPGVGVAPGVGVAPGVG SEQ ID NO: 1646
502-519


 


MMP9 + 12
LVPGVGVAPGVGVAPGVGVAPGVG SEQ ID NO: 1647
502-525


 


MMP9 + 12
LVPGVGVAPGVGVAPGVGVAPGVGLAPGVGVAPGVG
502-537



SEQ ID NO: 1648


 


MMP9 + 12
PGVGVAPGVGVAPG SEQ ID NO: 1649
504-517


 


MMP9 + 12
VGVAPGVGVAPGVGV SEQ ID NO: 1650
506-520


 


MMP9 + 12
VGVAPGVGVAPGVGVAPGVG SEQ ID NO: 1651
506-525


 


MMP9 + 12
VGVAPGVGVAPGVGVAPGVGLAPGVGVAPG SEQ ID NO: 1652
506-535


 


MMP9 + 12
VAPGVGVAPGVGVAPG SEQ ID NO: 1653
508-523


 


MMP9 + 12
VAPGVGVAPGVGVAPGVG SEQ ID NO: 1654
508-525


 


MMP9 + 12
VAPGVGVAPGVGVAPGVGLAPGVG SEQ ID NO: 1655
508-531


 


MMP9 + 12
VAPGVGVAPGVGVAPGVGLAPGVGVAPG SEQ ID NO: 1656
508-535


 


MMP9 + 12
VGVAPGVGVAPGVGLA SEQ ID NO: 1657
512-527


 


MMP9 + 12
VGVAPGVGVAPGVGLAPGVGVAPG SEQ ID NO: 1658
512-535


 


MMP9 + 12
VGVAPGVGVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG
512-552



SEQ ID NO: 1659


 


MMP9 + 12
VAPGVGVAPGVGLAPGVGVAPGVG SEQ ID NO: 1660
514-537


 


MMP9 + 12
VAPGVGVAPGVGLAPGVGVAPGVGVA SEQ ID NO: 1661
514-539


 


MMP9 + 12
VAPGVGVAPGVGLAPGVGVAPGVGVAPG SEQ ID NO: 1662
514-541


 


MMP9 + 12
VAPGVGVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGP
514-550



SEQ ID NO: 1663


 


MMP9 + 12
VAPGVGVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG
514-552



SEQ ID NO: 1664


 


MMP9 + 12
PGVGVAPGVGLAPG SEQ ID NO: 1665
516-529


 


MMP9 + 12
PGVGVAPGVGLAPGVGVAP SEQ ID NO: 1666
516-534


 


MMP9 + 12
PGVGVAPGVGLAPGVGVAPGVG SEQ ID NO: 1667
516-537


 


MMP9 + 12
VGVAPGVGLAPGVGVA SEQ ID NO: 1668
518-533


 


MMP9 + 12
VGVAPGVGLAPGVGVAP SEQ ID NO: 1669
518-534


 


MMP9 + 12
VGVAPGVGLAPGVGVAPGVGVAPG SEQ ID NO: 1670
518-541


 


MMP9 + 12
VGVAPGVGLAPGVGVAPGVGVAPGVG SEQ ID NO: 1671
518-543


 


MMP9 + 12
VGVAPGVGLAPGVGVAPGVGVAPGVGVAPG SEQ ID NO: 1672
518-547


 


MMP9 + 12
VGVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG
518-552



SEQ ID NO: 1673


 


MMP9 + 12
GVAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG
519-552



SEQ ID NO: 1674


 


MMP9 + 12
VAPGVGLAPGVGVA SEQ ID NO: 1675
520-533


 


MMP9 + 12
VAPGVGLAPGVGVAPG SEQ ID NO: 1676
520-535


 


MMP9 + 12
VAPGVGLAPGVGVAPGVG SEQ ID NO: 1677
520-537


 


MMP9 + 12
VAPGVGLAPGVGVAPGVGVA SEQ ID NO: 1678
520-539


 


MMP9 + 12
VAPGVGLAPGVGVAPGVGVAPG SEQ ID NO: 1679
520-541


 


MMP9 + 12
VAPGVGLAPGVGVAPGVGVAPGVGVA SEQ ID NO: 1680
520-545


 


MMP9 + 12
VAPGVGLAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO: 1681
520-552


 


MMP9 + 12
PGVGLAPGVGVAPG SEQ ID NO: 1682
522-535


 


MMP9 + 12
GVGLAPGVGVAPGVGVAPG SEQ ID NO: 1683
523-541


 


MMP9 + 12
VGLAPGVGVAPGVG SEQ ID NO: 1684
524-537


 


MMP9 + 12
VGLAPGVGVAPGVGVAPG SEQ ID NO: 1685
524-541


 


MMP9 + 12
VGLAPGVGVAPGVGVAPGVGVAPGIG SEQ ID NO: 1686
524-549


 


MMP9 + 12
VGLAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO: 1687
524-552


 


MMP9 + 12
VGLAPGVGVAPGVGVAPGVGVAPGIGPG SEQ ID NO: 1688
524-553


 


MMP9 + 12
VGLAPGVGVAPGVGVAPGVGVAPGIGPGGVAAA SEQ ID NO: 1689
524-556


 


MMP9 + 12
LAPGVGVAPGVGVAPGVG SEQ ID NO: 1690
526-543


 


MMP9 + 12
LAPGVGVAPGVGVAPGVGVA SEQ ID NO: 1691
526-545


 


MMP9 + 12
LAPGVGVAPGVGVAPGVGVAPGIGP SEQ ID NO: 1692
526-550


 


MMP9 + 12
LAPGVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO: 1693
526-552


 


MMP9 + 12
GVGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO: 1694
529-552


 


MMP9 + 12
VGVAPGVGVAPGVGVA SEQ ID NO: 1695
530-545


 


MMP9 + 12
VGVAPGVGVAPGVGVAPG SEQ ID NO: 1696
530-547


 


MMP9 + 12
VGVAPGVGVAPGVGVAPGIGPG SEQ ID NO: 1697
530-551


 


MMP9 + 12
VGVAPGVGVAPGVGVAPGIGPGG SEQ ID NO: 1698
530-552


 


MMP9 + 12
VGVAPGVGVAPGVGVAPGIGPGGVAAA SEQ ID NO: 1699
530-556


 


MMP9 + 12
VAPGVGVAPGVGVAP SEQ ID NO: 1700
532-546


 


MMP9 + 12
VAPGVGVAPGVGVAPGIG SEQ ID NO: 1701
532-549


 


MMP9 + 12
VAPGVGVAPGVGVAPGIGPGG SEQ ID NO: 1702
532-552


 


MMP9 + 12
PGVGVAPGVGVAPGIGPG SEQ ID NO: 1703
534-551


 


MMP9 + 12
PGVGVAPGVGVAPGIGPGG SEQ ID NO: 1704
534-552


 


MMP9 + 12
VGVAPGVGVAPGIGPGG SEQ ID NO: 1705
536-552


 


MMP9 + 12
VGVAPGVGVAPGIGPGGVAA SEQ ID NO: 1706
536-555


 


MMP9 + 12
VAPGVGVAPGIGPG SEQ ID NO: 1707
538-551


 


MMP9 + 12
PGVGVAPGIGPG SEQ ID NO: 1708
540-551


 


MMP9 + 12
VGVAPGIGPGGVAA SEQ ID NO: 1709
542-555


 


MMP9 + 12
PGGVAAAAK SEQ ID NO: 1710
550-558


 


MMP9 + 12
LRAAAGL SEQ ID NO: 1711
569-575


 


MMP9 + 12
LRAAAGLG SEQ ID NO: 1712
569-576


 


MMP9 + 12
LRAAAGLGA SEQ ID NO: 1713
569-577


 


MMP9 + 12
AAAGLGAGIPGLGVG SEQ ID NO: 1714
571-585


 


MMP9 + 12
AAAGLGAGIPGLGVGVG SEQ ID NO: 1715
571-587


 


MMP9 + 12
LGAGIPGLGVG SEQ ID NO: 1716
575-585


 


MMP9 + 12
LGAGIPGLGVGVG SEQ ID NO: 1717
575-587


 


MMP9 + 12
LGAGIPGLGVGVGVPGLGVG SEQ ID NO: 1718
575-594


 


MMP9 + 12
LGAGIPGLGVGVGVPG SEQ ID NO: 1719
575-590


 


MMP9 + 12
LGAGIPGLGVGVGVPGLGVGA SEQ ID NO: 1720
575-595


 


MMP9 + 12
LGAGIPGLGVGVGVPGL SEQ ID NO: 1721
575-591


 


MMP9 + 12
LGAGIPGLGVGVGVPGLG SEQ ID NO: 1722
575-592


 


MMP9 + 12
LGAGIPGLGVGVGVPGLGVGAGVPG SEQ ID NO: 1723
575-599


 


MMP9 + 12
LGAGIPGLGVGVGVPGLGVGAGVPGLG SEQ ID NO: 1724
575-601


 


MMP9 + 12
LGAGIPGLGVGVGVPGLGVGAGVPGLGVG SEQ ID NO: 1725
575-603


 


MMP9 + 12
LGAGIPGLGVGVGVPGLGVGAGVPGLGVGAGVPGFG
575-610



SEQ ID NO: 1726


 


MMP9 + 12
GAGIPGLGVGVGVPGLG SEQ ID NO: 1727
576-592


 


MMP9 + 12
AGIPGLGVGVGVPG SEQ ID NO: 1728
577-590


 


MMP9 + 12
GIPGLGVGVGVPGLGVGA SEQ ID NO: 1729
578-595


 


MMP9 + 12
LGVGVGVPGLGVGA SEQ ID NO: 1730
582-595


 


MMP9 + 12
VGVPGLGVGAGVPG SEQ ID NO: 1731
586-599


 


MMP9 + 12
VGVPGLGVGAGVPGL SEQ ID NO: 1732
586-600


 


MMP9 + 12
VGVPGLGVGAGVPGLG SEQ ID NO: 1733
586-601


 


MMP9 + 12
VGVPGLGVGAGVPGLGVG SEQ ID NO: 1734
586-603


 


MMP9 + 12
VGVPGLGVGAGVPGLGVGA SEQ ID NO: 1735
586-604


 


MMP9 + 12
VGAGVPGLGVGAGVPGFG SEQ ID NO: 1736
593-610


 


MMP9 + 12
PGALAAAK SEQ ID NO: 1737
646-653


 


MMP9 + 12
AKYGAAVPGVLGGLGA SEQ ID NO: 1738
655-670


 


MMP9 + 12
YGAAVPGVLGG SEQ ID NO: 1739
657-667


 


MMP9 + 12
YGAAVPGVLGGLG SEQ ID NO: 1740
657-669


 


MMP9 + 12
YGAAVPGVLGGLGA SEQ ID NO: 1741
657-670


 


MMP9 + 12
YGAAVPGVLGGLGALG SEQ ID NO: 1742
657-672


 


MMP9 + 12
YGAAVPGVLGGLGALGGVGIPGG SEQ ID NO: 1743
657-679


 


MMP9 + 12
YGAAVPGVLGGLGALGGVGIPGGVVGAGPAA SEQ ID NO: 1744
657-687


 


MMP9 + 12
GAAVPGVLGGLG SEQ ID NO: 1745
658-669


 


MMP9 + 12
GAAVPGVLGGLGALGGVGIPGG SEQ ID NO: 1746
658-679


 


MMP9 + 12
AVPGVLGGLGA SEQ ID NO: 1747
660-670


 


MMP9 + 12
AVPGVLGGLGALGGVGIPGG SEQ ID NO: 1748
660-679


 


MMP9 + 12
VLGGLGALGGVGIPGG SEQ ID NO: 1749
664-679


 


MMP9 + 12
GGLGALGGVGIPGGVVGAGPA SEQ ID NO: 1750
666-686


 


MMP9 + 12
GGLGALGGVGIPGGVVGAGPAAA SEQ ID NO: 1751
666-688


 


MMP9 + 12
LGALGGVGIPGG SEQ ID NO: 1752
668-379


 


MMP9 + 12
LGALGGVGIPGGVVGAGPA SEQ ID NO: 1753
668-686


 


MMP9 + 12
LGALGGVGIPGGVVGAGPAA SEQ ID NO: 1754
668-687


 


MMP9 + 12
LGALGGVGIPGGVVGAGPAAA SEQ ID NO: 1755
668-688


 


MMP9 + 12
LGALGGVGIPGGVVGAGPAAAA SEQ ID NO: 1756
668-689


 


MMP9 + 12
ALGGVGIPGGVVGAGPAA SEQ ID NO: 1757
670-687


 


MMP9 + 12
ALGGVGIPGGVVGAGPAAA SEQ ID NO: 1758
670-688


 


MMP9 + 12
LGGVGIPGGV SEQ ID NO: 1759
671-680


 


MMP9 + 12
LGGVGIPGGVVGAGPA SEQ ID NO: 1760
671-686


 


MMP9 + 12
LGGVGIPGGVVGAGPAAA SEQ ID NO: 1761
671-688


 


MMP9 + 12
LGGVGIPGGVVGAGPAAAAA SEQ ID NO: 1762
671-690


 


MMP9 + 12
LGGVGIPGGVVGAGPAAAAAAAK SEQ ID NO: 1763
671-693


 


MMP9 + 12
GVGIPGGVVGAGPAAAA SEQ ID NO: 1764
673-689


 


MMP9 + 12
GVGIPGGVVGAGPAAAAAAAK SEQ ID NO: 1765
673-693


 


MMP9 + 12
VGIPGGVVGAGPAAA SEQ ID NO: 1766
674-688


 


MMP9 + 12
VGIPGGVVGAGPAAAAAAAK SEQ ID NO: 1767
674-693


 


MMP9 + 12
IPGGVVGAGPAAAA SEQ ID NO: 1768
676-689


 


MMP9 + 12
VVGAGPAAAAAAAK SEQ ID NO: 1769
680-693


 


MMP9 + 12
VGAGPAAAAAAAK SEQ ID NO: 1770
681-693


 


MMP9 + 12
AGPAAAAAAAK SEQ ID NO: 1771
683-693


 


MMP9 + 12
GPAAAAAAAK SEQ ID NO: 1772
684-693


 


MMP9 + 12
PAAAAAAAK SEQ ID NO: 1773
685-693


 


MMP9 + 12
FGLVGAAGLGGLGVGGLGVPGVGG SEQ ID NO: 1774
701-724


 


MMP9 + 12
GLVGAAGLGGLG SEQ ID NO: 1775
702-713


 


MMP9 + 12
GLVGAAGLGGLGVGG SEQ ID NO: 1776
702-716


 


MMP9 + 12
GLVGAAGLGGLGVGGLGVPGVG SEQ ID NO: 1777
702-723


 


MMP9 + 12
GLVGAAGLGGLGVGGLGVPGVGG SEQ ID NO: 1778
702-724


 


MMP9 + 12
LVGAAGLGGLGVG SEQ ID NO: 1779
703-715


 


MMP9 + 12
LVGAAGLGGLGVGG SEQ ID NO: 1780
703-716


 


MMP9 + 12
LVGAAGLGGLGVGGL SEQ ID NO: 1781
703-717


 


MMP9 + 12
LVGAAGLGGLGVGGLGVPGVGGLG SEQ ID NO: 1782
703-726


 


MMP9 + 12
LVGAAGLGGLGVGGLGVPGVGGLGGIPPAAA SEQ ID NO: 1783
703-733


 


MMP9 + 12
VGAAGLGGLGVGG SEQ ID NO: 1784
704-716


 


MMP9 + 12
LGGLGVGGLGVPG SEQ ID NO: 1785
709-721


 


MMP9 + 12
LGGLGVGGLGVPGVG SEQ ID NO: 1786
709-723


 


MMP9 + 12
LGGLGVGGLGVPGVGGL SEQ ID NO: 1787
709-725


 


MMP9 + 12
LGGLGVGGLGVPGVGGLG SEQ ID NO: 1788
709-726


 


MMP9 + 12
LGVGGLGVPGVGGLG SEQ ID NO: 1789
712-726


 


MMP9 + 12
GLGVPGVGGLGGIPPAAAAK SEQ ID NO: 1790
716-735


 


MMP9 + 12
LGGIPPAAAAK SEQ ID NO: 1791
725-735


 


MMP9 + 12
LGGVLGGAGQFPL SEQ ID NO: 1792
744-756


 


MMP9 + 12
LGGVLGGAGQFPLGGVAAR SEQ ID NO: 1793
744-762


 


MMP9 + 12
LGGVLGGAGQFPLGGVAARPG SEQ ID NO: 1794
744-764


 


MMP9 + 12
LGGVLGGAGQFPLGGVAARPGFG SEQ ID NO: 1795
744-766


 


MMP9 + 12
GGVLGGAGQFPLGGVAARPG SEQ ID NO: 1796
745-764


 


MMP9 + 12
GAGQFPLGGVAAR SEQ ID NO: 1797
750-762


 


MMP9 + 12
GAGQFPLGGVAARPGFG SEQ ID NO: 1798
750-766


 


MMP9 + 12
AGQFPLGGVAARPGFG SEQ ID NO: 1799
751-766


 


MMP9 + 12
FPLGGVAARPG SEQ ID NO: 1800
754-764


 


MMP9 + 12
PLGGVAAR SEQ ID NO: 1801
755-762


 


MMP9 + 12
PLGGVAARPG SEQ ID NO: 1802
755-764


 


MMP9 + 12
PLGGVAARPGFG SEQ ID NO: 1803
755-766


 


MMP9 + 12
PLGGVAARPGFGL SEQ ID NO: 1804
755-767


 


MMP9 + 12
PLGGVAARPGFGLSPIFPG SEQ ID NO: 1805
755-773


 


MMP9 + 12
LGGVAAR SEQ ID NO: 1806
756-762


 


MMP9 + 12
LGGVAARP SEQ ID NO: 1807
756-763


 


MMP9 + 12
LGGVAARPG SEQ ID NO: 1808
756-764


 


MMP9 + 12
LGGVAARPGF SEQ ID NO: 1809
756-765


 


MMP9 + 12
LGGVAARPGFG SEQ ID NO: 1810
756-766


 


MMP9 + 12
LGGVAARPGFGL SEQ ID NO: 1811
756-767


 


MMP9 + 12
LGGVAARPGFGLSP SEQ ID NO: 1812
756-769


 


MMP9 + 12
LGGVAARPGFGLSPIFPG SEQ ID NO: 1813
756-773


 


MMP9 + 12
LGGVAARPGFGLSPIFPGG SEQ ID NO: 1814
756-774


 


MMP9 + 12
LGGVAARPGFGLSPIFPGGA SEQ ID NO: 1815
756-775


 


MMP9 + 12
GGVAARPGFG SEQ ID NO: 1816
757-766


 


MMP9 + 12
GGVAARPGFGL SEQ ID NO: 1817
757-767


 


MMP9 + 12
GGVAARPGFGLSPIFPGGA SEQ ID NO: 1818
757-775


 


MMP9 + 12
GVAARPGFGLSPIF SEQ ID NO: 1819
758-771


 


MMP9 + 12
GVAARPGFGLSPIFP SEQ ID NO: 1820
758-772


 


MMP9 + 12
VAARPGFG SEQ ID NO: 1821
759-766


 


MMP9 + 12
VAARPGFGLSPIFP SEQ ID NO: 1822
759-772


 


MMP9 + 12
VAARPGFGLSPIFPG SEQ ID NO: 1823
759-773


 


MMP9 + 12
RPGFGLSPIFPG SEQ ID NO: 1824
762-773


 


MMP9 + 12
PGFGLSPIFPGG SEQ ID NO: 1825
763-774


 


MMP9 + 12
PGFGLSPIFPGGA SEQ ID NO: 1826
763-775


 


ADAMTS-1
P.GVGLPGVYPGGVLPGAR.F SEQ ID NO: 1827
143-159


 


ADAMTS-1
G.VGLPGVYPGGVLPGAR.F SEQ ID NO: 1828
144-159


 


ADAMTS-1
G.LPGVYPGGVLPGAR.F SEQ ID NO: 1829
146-159


 


ADAMTS-1
P.GVYPGGVLPGAR.F SEQ ID NO: 1830
148-159


 


ADAMTS-1
K.AGYPTGTGVGPQAAAAAAAK.A SEQ ID NO: 1831
242-261


 


ADAMTS-1
G.GPGFGPGVVGVPGAGVPGVGVPGA.G SEQ ID NO: 1832
326-349


 


ADAMTS-1
G.FGPGVVGVPGAGVPGVGVPG.A SEQ ID NO: 1833
329-348


 


ADAMTS-1
F.GPGVVGVPGAGVPGVGVPG.A SEQ ID NO: 1834
330-348


 


ADAMTS-1
G.VPGVGVPGAGIPVVPG.A SEQ ID NO: 1835
341-356


 


ADAMTS-1
G.ARPGVGVGGIPTYGVG.A SEQ ID NO: 1836
385-400


 


ADAMTS-1
G.ARPGVGVGGIPTYGVGAGG.F SEQ ID NO: 1837
385-403


 


ADAMTS-1
A.RPGVGVGGIPTYGVGAG.G SEQ ID NO: 1838
386-402


 


ADAMTS-1
G.GVPGVGGVPGVGISPEAQAAAA.A SEQ ID NO: 1839
425-446


 


ADAMTS-1
G.VPGVGISPEAQAAAAAK.A SEQ ID NO: 1840
432-448


 


ADAMTS-1
G.VGISPEAQAAAAAK.A SEQ ID NO: 1841
435-448


 


ADAMTS-1
V.PGVGVAPGVGVAPGVGVAPGVGL.A SEQ ID NO: 1842
504-526


 


ADAMTS-1
G.VAPGVGVAPGVGVAPGVGLAPGVGVAPG.V SEQ ID NO: 1843
508-535


 


ADAMTS-1
G.VGVAPGVGVAPGVGLAPGVG.V SEQ ID NO: 1844
512-531


 


ADAMTS-1
G.VGVAPGVGVAPGVGLAPGVGVAPGVG.V SEQ ID NO: 1845
512-537


 


ADAMTS-1
A.PGVGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1846
528-551


 


ADAMTS-1
G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1847
532-551


 


ADAMTS-1
G.AAVPGVLGGLGALGGVGIPG.G SEQ ID NO: 1848
659-678


 


ADAMTS-1
G.AAGLGGLGVGGLGVPGVGGLG.G SEQ ID NO: 1849
706-726


 


ADAMTS-4
P.GVGLPGVYPGGVLPGAR.F SEQ ID NO: 1827
143-159


 


ADAMTS-4
G.LPGVYPGGVLPGAR.F SEQ ID NO: 1829
146-159


 


ADAMTS-4
K.AGYPTGTGVGPQAAAAAAAK.A SEQ ID NO: 1831
242-261


 


ADAMTS-4
G.GAGVPGVPGAIPGIGGIAGVG.T SEQ ID NO: 1850
279-299


 


ADAMTS-4
G.AGVPGVPGAIPGIGGIAGVG.T SEQ ID NO: 1851
280-299


 


ADAMTS-4
A.GVGTPAAAAAAAAAAK.A SEQ ID NO: 1852
297-312


 


ADAMTS-4
G.VGTPAAAAAAAAAAK.A SEQ ID NO: 1853
298-312


 


ADAMTS-4
G.GPGFGPGVVGVPGAGVPGVGVPG.A SEQ ID NO: 1854
326-348


 


ADAMTS-4
G.ARPGVGVGGIPTYGVGA.G SEQ ID NO: 1855
385-401


 


ADAMTS-4
A.RPGVGVGGIPTYGVGAG.G SEQ ID NO: 1838
386-402


 


ADAMTS-4
A.RPGVGVGGIPTYGVGAGG.F SEQ ID NO: 1856
386-403


 


ADAMTS-4
G.VGISPEAQAAAAAK.A SEQ ID NO: 1841
435-448


 


ADAMTS-4
G.VGVAPGVGVAPGVGVAPGVGLAPGVG.V SEQ ID NO: 1857
506-531


 


ADAMTS-4
A.PGVGVAPGVGLAPGVGVAPGVGVA.P SEQ ID NO: 1858
516-539


 


ADAMTS-4
G.VGVAPGVGLAPGVGVAPGVG.V SEQ ID NO: 1859
518-537


 


ADAMTS-4
L.APGVGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1860
527-551


 


ADAMTS-4
Y.GAAVPGVLGGLGALGGVGIPG.G SEQ ID NO: 1861
658-678


 


ADAMTS-4
G.AAVPGVLGGLGALGGVGIPG.G SEQ ID NO: 1848
659-678


 


ADAMTS-4
G.GAGQFPLGGVAARPGFGL.S SEQ ID NO: 1862
750-767


 


ADAMTS-8
L.VPGGVADAAAAYK.A SEQ ID NO: 1863
092-104


 


ADAMTS-8
G.VGLPGVYPGGVLPGAR.F SEQ ID NO: 1828
144-159


 


ADAMTS-8
G.LPGVYPGGVLPGAR.F SEQ ID NO: 1829
146-159


 


ADAMTS-8
P.GVYPGGVLPGAR.F SEQ ID NO: 1830
148-159


 


ADAMTS-8
V.YPGGVLPGAR.F SEQ ID NO: 1864
150-159


 


ADAMTS-8
F.GPGVVGVPGAGVPGVGVPG.A SEQ ID NO: 1834
330-348


 


ADAMTS-8
G.ARPGVGVGGIPTYGVGA.G SEQ ID NO: 1855
385-401


 


ADAMTS-8
V.APGVGVAPGVGVAPGVGLAPGVGV.A SEQ ID NO: 1865
509-532


 


ADAMTS-8
L.APGVGVAPGVGVAPGVGV.A SEQ ID NO: 1866
527-544


 


ADAMTS-8
L.APGVGVAPGVGVAPGVGVAPG.I SEQ ID NO: 1867
527-547


 


ADAMTS-8
L.APGVGVAPGVGVAPGVGVAPGIG.P SEQ ID NO: 1868
527-549


 


ADAMTS-8
L.APGVGVAPGVGVAPGVGVAPGIGP.G SEQ ID NO: 1869
527-550


 


ADAMTS-8
L.APGVGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1860
527-551


 


ADAMTS-8
L.APGVGVAPGVGVAPGVGVAPGIGPGGVAA.A SEQ ID NO: 1870
527-555


 


ADAMTS-8
G.VGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1871
530-551


 


ADAMTS-8
G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1847
532-551


 


ADAMTS-8
G.AAVPGVLGGLGALGGVGIPG.G SEQ ID NO: 1848
659-678


 


ADAMTS-8
G.AAVPGVLGGLGALGGVGIPGG.V SEQ ID NO: 1872
659-679


 


ADAMTS-8
A.AVPGVLGGLGALGGVGIPG.G SEQ ID NO: 1873
660-678


 


ADAMTS-8
A.VPGVLGGLGALGGVGIPGG.V SEQ ID NO: 1874
661-679


 


ADAMTS-8
A.GQFPLGGVAARPGFGL.S SEQ ID NO: 1875
752-767


 


Cat K
G.ALVPGGVADAAAAYK.A SEQ ID NO: 1876
090-104


 


Cat K
G.LPYTTGKLPYGYGPG.G SEQ ID NO: 1877
219-233


 


Cat K
A.AAAAAAKAAAKFGA.G SEQ ID NO: 1878
255-268


 


Cat K
A.GVGTPAAAAAAAAAAK.A SEQ ID NO: 1852
297-312


 


Cat K
A.AAAAAAAAAKAAKYGA.A SEQ ID NO: 1879
303-318


 


Cat K
G.FGPGVVGVPGAGVPGVGVPG.A SEQ ID NO: 1833
329-348


 


Cat K
G.VGISPEAQAAAAAK.A SEQ ID NO: 1841
435-448


 


Cat K
G.VAPGVGVAPGVGVAPGVGLAPGVG.V SEQ ID NO: 1880
508-531


 


Cat K
G.VGVAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1871
530-551


 


Cat K
G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1847
532-551


 


Cat S
T.FPGALVPGGVADAAAAYK.A SEQ ID NO: 1881
087-104


 


Cat S
G.VGLPGVYPGGVLPGAR.F SEQ ID NO: 1828
144-159


 


Cat S
G.LPGVYPGGVLPGARFPGVG.V SEQ ID NO: 1882
146-164


 


Cat S
G.YPTGTGVGPQAAAAAAAK.A SEQ ID NO: 1883
244-261


 


Cat S
G.GAGVPGVPGAIPGIGGIAGVG.T SEQ ID NO: 1850
279-299


 


Cat S
G.TPAAAAAAAAAAKAAK.Y SEQ ID NO: 1884
300-315


 


Cat S
G.VPGAGVPGVGVPGAGIPVVP.G SEQ ID NO: 1885
336-355


 


Cat S
G.VPGAGVPGVGVPGAGIPVVPGAGIPG.A SEQ ID NO: 1886
336-361


 


Cat S
G.ISPEAQAAAAAKAAK.Y SEQ ID NO: 1887
437-451


 


Cat S
V.PGVGVAPGVGVAPGVGVA.P SEQ ID NO: 1888
504-521


 


Cat S
G.VAPGVGVAPGVGVAPGIGPGGVA.A SEQ ID NO: 1889
532-554


 


Cat S
G.IPGGVVGAGPAAAAAAAK.A SEQ ID NO: 1890
676-693


 


MMP1
G.GVLPGARFPGVGVLPGVPTGA.G SEQ ID NO: 1891
153-173


 


MMP1
G.GVPGVGGVPGVGISPEA.Q SEQ ID NO: 1892
425-441


 


MMP1
V.PGVGVAPGVGVAPGVGVA.P SEQ ID NO: 1888
504-521


 


MMP1
G.VGVAPGVGVAPGVGVAPGVG.L SEQ ID NO: 1893
506-525


 


MMP1
G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1847
532-551


 


MMP1
A.AVPGVLGGLGALGGVGIPG.G SEQ ID NO: 1873
660-678


 


MMP1


 


MMP3
G.ALVPGGVADAAAAYK.A SEQ ID NO: 1876
090-104


 


MMP3
G.YPTGTGVGPQAAAAAAAK.A SEQ ID NO: 1883
244-261


 


MMP3
G.VPGVPGAIPGIGGIAGVG.T SEQ ID NO: 1894
282-299


 


MMP3
F.GPGVVGVPGAGVPGVGVPGA.G SEQ ID NO: 1895
330-349


 


MMP3
G.VGISPEAQAAAAAK.A SEQ ID NO: 1841
435-448


 


MMP3
G.VGVAPGVGVAPGVGLAPGVG.V SEQ ID NO: 1844
512-531


 


MMP3
G.VAPGVGVAPGVGVAPGIGPG.G SEQ ID NO: 1847
532-551


 


MMP8
P.GVYPGGVLPGAR.F SEQ ID NO: 1830
148-159


 


MMP8
K.AGYPTGTGVGPQAAAAAAAK.A SEQ ID NO: 1831
242-261


 


MMP8
G.VPGVPGAIPGIGGIAGVG.T SEQ ID NO: 1894
282-299


 


MMP8
F.GPGVVGVPGAGVPGVGVPG.A SEQ ID NO: 1834
330-348


 


MMP8
G.VPGVGVPGAGIPVVPGA.G SEQ ID NO: 1896
341-357


 


MMP8
G.ARPGVGVGGIPTYGVG.A SEQ ID NO: 1836
385-400


 


MMP8
A.RPGVGVGGIPTYGVGAG.G SEQ ID NO: 1838
386-402


 


MMP8
G.VGVAPGVGVAPGVGVAP.G SEQ ID NO: 1897
506-522


 


MMP8
G.VGVAPGVGVAPGVGLAPGVG.V SEQ ID NO: 1844
512-531


 


MMP8
G.VGVAPGVGVAPGVGVAP.G SEQ ID NO: 1897
530-546


 


MMP8
G.IPGGVVGAGPAAAAAAAK.A SEQ ID NO: 1890
676-693





*Aminoacid residue numbers in the human elastin sequence






Accordingly, in a method of the invention, said peptide fragments preferably comprise a neo-epitope formed by cleavage of elastin by a protease at an N- or C-terminal site, or where indicated a site marked by the sign in any one of the partial sequences of elastin in Table 24.
The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neo-epitope formed by cleavage of elastin.
Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:







TABLE 25





N-terminal sequences of protease generated


peptide fragments of elastin.

















GVPGAI SEQ ID NO: 1898
AIPGGV SEQ ID NO: 1899
GVPGGV SEQ ID NO: 1900


 


ALGGGA SEQ ID NO: 1901
LGGGAL SEQ ID NO: 1902
GGALGP SEQ ID NO: 1903


 


PLKPVP SEQ ID NO: 1904
LKPVPG SEQ ID NO: 1905
GLAGAG SEQ ID NO: 1906


 


GLGAGL SEQ ID NO: 1907
LGAGLG SEQ ID NO: 1908
AGLGAF SEQ ID NO: 1909


 


LVPGGV SEQ ID NO: 1910
VADAAA SEQ ID NO: 1911
KAAKAG SEQ ID NO: 1912


 


PVGYPG SEQ ID NO: 1913
ARFPGV SEQ ID NO: 1914
RFPGVG SEQ ID NO: 1915


 


VGPFGG SEQ ID NO: 1916
GPQPGV SEQ ID NO: 1917
PQPGVP SEQ ID NO: 1918


 


TTGKLP SEQ ID NO: 1919
LPYGYG SEQ ID NO: 1920
GYGPGG SEQ ID NO: 1921


 


FGAGAA SEQ ID NO: 1922
GVLPGV SEQ ID NO: 1923
VLPGVG SEQ ID NO: 1924


 


AGIPGL SEQ ID NO: 1925
VPGAIP SEQ ID NO: 1926
AIPGIG SEQ ID NO: 1927


 


TPAAAA SEQ ID NO: 1928
PAAAAA SEQ ID NO: 1929
AAAAAA SEQ ID NO: 1930


 


VGVPGA SEQ ID NO: 1931
AVGPGV SEQ ID NO: 1932
GVPGVG SEQ ID NO: 1933


 


GIPVVP SEQ ID NO: 1934
IPGAAV SEQ ID NO: 1935
GAAVPG SEQ ID NO: 1936


 


ARPGVG SEQ ID NO: 1937
RPGVGV SEQ ID NO: 1938
VGGIPT SEQ ID NO: 1939


 


SVGGVP SEQ ID NO: 1940
VGGVPG SEQ ID NO: 1941
GVGTPA SEQ ID NO: 1942


 


VGVAPG SEQ ID NO: 1943
VAPGVG SEQ ID NO: 1944
GVAPGV SEQ ID NO: 1945


 


GVPVAP SEQ ID NO: 1946
GAGIPG SEQ ID NO: 1947
PGGVAA SEQ ID NO: 1948


 


LGAGIP SEQ ID NO: 1949
PGFGPG SEQ ID NO: 1950
PGVVGV SEQ ID NO: 1951


 


PGALAA SEQ ID NO: 1952
AKYGAA SEQ ID NO: 1953
YGAAVP SEQ ID NO: 1954


 


ALGGVG SEQ ID NO: 1955
LGGVGI SEQ ID NO: 1956
GVGIPG SEQ ID NO: 1957


 


AGPAAA SEQ ID NO: 1958
GPAAAA SEQ ID NO: 1959
FGLVGA SEQ ID NO: 1960


 


GLGVPG SEQ ID NO: 1961
LGGIPP SEQ ID NO: 1962
LGGVLG SEQ ID NO: 1963


 


PLGGVA SEQ ID NO: 1964
LGGVAA SEQ ID NO: 1965
GGVAAR SEQ ID NO: 1966


 


GVGLPG SEQ ID NO: 1967
VGLPGV SEQ ID NO: 1968
LPGVYP SEQ ID NO: 1969


 


VPGVPV SEQ ID NO: 1970
VPGVGI SEQ ID NO: 1971
VGISPE SEQ ID NO: 1972


 


APGVGV SEQ ID NO: 1973
VPGGVA SEQ ID NO: 1974
YPGGVL SEQ ID NO: 1975


 


GPGFGP SEQ ID NO: 1976
YPTGTG SEQ ID NO: 1977
VPGAGV SEQ ID NO: 1978


 


VPGGVF SEQ ID NO: 1979
GVFYPG SEQ ID NO: 1980
VFYPGA SEQ ID NO: 1981


 


LGPGGK SEQ ID NO: 1982
GPGGKP SEQ ID NO: 1983
PGGKPL SEQ ID NO: 1984


 


LAGAGL SEQ ID NO: 1985
AGAGLG SEQ ID NO: 1986
GAGLGA SEQ ID NO: 1987


 


LGAFPA SEQ ID NO: 1988
AFPAVT SEQ ID NO: 1989
AVTFPG SEQ ID NO: 1990


 


LGVSAG SEQ ID NO: 1991
VPGVGL SEQ ID NO: 1992
PGVGLP SEQ ID NO: 1993


 


FPGVGV SEQ ID NO: 1994
KPGAPG SEQ ID NO: 1995
PKAPGV SEQ ID NO: 1493


 


PGVPLG SEQ ID NO: 1996
GYPIKA SEQ ID NO: 1997
PKLPGG SEQ ID NO: 1998


 


YGPGGV SEQ ID NO: 1999
AGYPTG SEQ ID NO: 2000
TGVGPQ SEQ ID NO: 2001


 


GAGVPG SEQ ID NO: 2002
AGVPGV SEQ ID NO: 2003
GVPGVP SEQ ID NO: 2004


 


IPGIGG SEQ ID NO: 2005
IGGIAG SEQ ID NO: 2006
GIAGVG SEQ ID NO: 2007


 


VPGVGV SEQ ID NO: 2008
VPVGVA SEQ ID NO: 2009
VPGAGI SEQ ID NO: 2010


 


AVPGVV SEQ ID NO: 2011
VPGVVS SEQ ID NO: 2012
YGARPG SEQ ID NO: 2013


 


GVGAGG SEQ ID NO: 2014
VGAGGF SEQ ID NO: 2015
VGVGGI SEQ ID NO: 2016


 


FGLVPG SEQ ID NO: 2017
GLVPGV SEQ ID NO: 2018
LVPGVG SEQ ID NO: 2019


 


PGVGLA SEQ ID NO: 2020
GVGLAP SEQ ID NO: 2021
VGLAPG SEQ ID NO: 2022


 


LRAAAG SEQ ID NO: 2023
LVGAAG SEQ ID NO: 2024
LVPGGP SEQ ID NO: 2025


 


GIPGLG SEQ ID NO: 2026
LGVGVG SEQ ID NO: 2027
VGVPGL SEQ ID NO: 2028


 


AAAGLG SEQ ID NO: 2029
AVPGVL SEQ ID NO: 2030
VLGGLG SEQ ID NO: 2031


 


VGIPGG SEQ ID NO: 2032
IPGGVV SEQ ID NO: 2033
VVGAGP SEQ ID NO: 2034


 


GLVGAA SEQ ID NO: 2035
VGAAGL SEQ ID NO: 2036
LGGLGV SEQ ID NO: 2037


 


GGVLGG SEQ ID NO: 2038
GAGQFP SEQ ID NO: 2039
AFQFPL SEQ ID NO: 2040


 


GVAARP SEQ ID NO: 2041
VAARPG SEQ ID NO: 2042
RPGFGL SEQ ID NO: 2043


 


GVYPGG SEQ ID NO: 2044
LPYTTG SEQ ID NO: 2045
FGPGVV SEQ ID NO: 2046


 


AAVPGV SEQ ID NO: 2047
AAGLGG SEQ ID NO: 2048
FPGALV SEQ ID NO: 2049


 


VPGVLG SEQ ID NO: 2050
GQFPLG SEQ ID NO: 2051
ALVPGG SEQ ID NO: 2052


 


ISPEAQ SEQ ID NO: 2053
GVLPGA SEQ ID NO: 2054
VGAGVP SEQ ID NO: 2055


 


LGALGG SEQ ID NO: 2056
VPGVPG SEQ ID NO: 2057
GGLGAL SEQ ID NO: 2058


 


GKPLKP SEQ ID NO: 2059
IAGVGT SEQ ID NO: 2060
VGAGPA SEQ ID NO: 2061


 


AGLGAG SEQ ID NO: 2062
VVGVPG SEQ ID NO: 2063
LGVGGL SEQ ID NO: 2064


 


VTFPGA SEQ ID NO: 2065
AGIPVV SEQ ID NO: 2066
FPLGGV SEQ ID NO: 2067


 


GLPGVY SEQ ID NO: 2068
GARPGV SEQ ID NO: 2069
PGFGLS SEQ ID NO: 2070


 


GAFAGI SEQ ID NO: 2071
VAGVPS SEQ ID NO: 2072
GPGVVG SEQ ID NO: 2073


 


YTTGKL SEQ ID NO: 2074
PGVGVA SEQ ID NO: 2075
VGTPAA SEQ ID NO: 2076


 


PQAAAA SEQ ID NO: 2077
LAPGVG SEQ ID NO: 2078









or with any of the following sequences at the C-terminal of a peptide:








TABLE 26





C-terminal sequences of protease generated


peptide fragments of Elastin.







Elastin









PGGVPG SEQ ID NO: 2079
PGAGLG SEQ ID NO: 2080
GAGLGA SEQ ID NO: 1987


 


GALGGG SEQ ID NO: 2081
LGGGAL SEQ ID NO: 1902
GGGALG SEQ ID NO: 2082


 


GLGAFP SEQ ID NO: 2083
LGAFPA SEQ ID NO: 1988
LGAGLG SEQ ID NO: 1908


 


VPGGVA SEQ ID NO: 1974
ADAAAA SEQ ID NO: 2084
PGVLGG SEQ ID NO: 2085


 


RFPGVG SEQ ID NO: 1915
VGVLPG SEQ ID NO: 2086
VPTGAG SEQ ID NO: 2087


 


PKAPGV SEQ ID NO: 1493
GVGPFG SEQ ID NO: 2088
VPLGYP SEQ ID NO: 2089


 


LPYGYG SEQ ID NO: 1920
AAAAAK SEQ ID NO: 2090
IPGIGG SEQ ID NO: 2005


 


GIAGVG SEQ ID NO: 2007
AIPGIG SEQ ID NO: 1927
IGGIAG SEQ ID NO: 2006


 


AAAAKA SEQ ID NO: 2091
VVGVPG SEQ ID NO: 2063
GVPGVG SEQ ID NO: 1933


 


GVGVPG SEQ ID NO: 2092
PVVPGA SEQ ID NO: 2093
VVSPEA SEQ ID NO: 2094


 


VGGIPT SEQ ID NO: 1939
GGIPTY SEQ ID NO: 2095
GIPTYG SEQ ID NO: 2096


 


GVGVGG SEQ ID NO: 2097
GVGGIP SEQ ID NO: 2098
VGVPGL SEQ ID NO: 2028


 


GFPGFG SEQ ID NO: 2099
FGVGVG SEQ ID NO: 2100
GVGAGG SEQ ID NO: 2014


 


PGVGIS SEQ ID NO: 2101
SPEAQA SEQ ID NO: 2102
VAPGVG SEQ ID NO: 1944


 


PGVGVA SEQ ID NO: 2075
GVGVAP SEQ ID NO: 2103
APGVGL SEQ ID NO: 2104


 


GIGPGG SEQ ID NO: 2105
APGIGP SEQ ID NO: 2106
VAPGIG SEQ ID NO: 2107


 


RAAAGL SEQ ID NO: 2108
AAAGLG SEQ ID NO: 2029
AAGLGA SEQ ID NO: 2109


 


GVPGLG SEQ ID NO: 2110
GVPGFG SEQ ID NO: 2111
AGVPGL SEQ ID NO: 2112


 


VLGGLG SEQ ID NO: 2031
VGIPGG SEQ ID NO: 2032
GAGPAA SEQ ID NO: 2113


 


PAAAAA SEQ ID NO: 1929
VPGVGG SEQ ID NO: 2114
GLGGLG SEQ ID NO: 2115


 


GLGVPG SEQ ID NO: 1961
PGVGGL SEQ ID NO: 2116
PAAAAK SEQ ID NO: 2117


 


RPGFGL SEQ ID NO: 2043
GVAARP SEQ ID NO: 2041
AARPGF SEQ ID NO: 2118


 


LSPIFP SEQ ID NO: 2119
AQAAAA SEQ ID NO: 2120
GPGIPG SEQ ID NO: 2121


 


GPGGVA SEQ ID NO: 2122
TPAAAA SEQ ID NO: 1928
PGGVAA SEQ ID NO: 1948


 


GLGALG SEQ ID NO: 2123
LGALGG SEQ ID NO: 2056
ALGPGG SEQ ID NO: 2124


 


GALGPG SEQ ID NO: 2125
VAPVGV SEQ ID NO: 2126
KPVPGG SEQ ID NO: 2127


 


AFPAVT SEQ ID NO: 1989
AVTFPG SEQ ID NO: 1990
VTFPGA SEQ ID NO: 2065


 


AAAAYK SEQ ID NO: 2128
AAKAGA SEQ ID NO: 2129
VPQPGA SEQ ID NO: 2130


 


PGVPTG SEQ ID NO: 2131
GVPTGA SEQ ID NO: 2132
AGVKPK SEQ ID NO: 2133


 


PIKAPK SEQ ID NO: 2134
KLPGGY SEQ ID NO: 2135
YGYGPG SEQ ID NO: 2136


 


VGTPAA SEQ ID NO: 2076
VPGVPG SEQ ID NO: 2057
GVGTPA SEQ ID NO: 1942


 


GIGGIA SEQ ID NO: 2137
GTPAAA SEQ ID NO: 2138
AAAAAA SEQ ID NO: 1930


 


IPVVPG SEQ ID NO: 2139
VGVPGA SEQ ID NO: 1931
GVPGAG SEQ ID NO: 2140


 


GAGIPG SEQ ID NO: 1947
PEAAAK SEQ ID NO: 2141
ARPGVG SEQ ID NO: 1937


 


PTYGVG SEQ ID NO: 2142
TYGVGA SEQ ID NO: 2143
YGVGAG SEQ ID NO: 2144


 


IPTYGV SEQ ID NO: 2145
PGAIPG SEQ ID NO: 2146
VGAGGF SEQ ID NO: 2015


 


AGGFPG SEQ ID NO: 2147
GIPGVA SEQ ID NO: 2148
GISPEA SEQ ID NO: 2149


 


VGVAPG SEQ ID NO: 1943
VGLAPG SEQ ID NO: 2022
VPGAPG SEQ ID NO: 2150


 


PGVGLA SEQ ID NO: 2020
LAPGVG SEQ ID NO: 2078
APGVGV SEQ ID NO: 1973


 


GVAPGV SEQ ID NO: 1945
GGVAAA SEQ ID NO: 2151
PGIGPG SEQ ID NO: 2152


 


GLGVGG SEQ ID NO: 2153
PGLGVG SEQ ID NO: 2154
LGVGVG SEQ ID NO: 2027


 


GLGVGA SEQ ID NO: 2155
ALAAAK SEQ ID NO: 2156
LGGLGA SEQ ID NO: 2157


 


VGAGPA SEQ ID NO: 2061
AGPAAA SEQ ID NO: 1958
GPAAAA SEQ ID NO: 1959


 


GGLGVG SEQ ID NO: 2158
LGVGGL SEQ ID NO: 2064
GVGGLG SEQ ID NO: 2159


 


AGQFPL SEQ ID NO: 2160
GGVAAR SEQ ID NO: 1966
VAARPG SEQ ID NO: 2042


 


GFGLSP SEQ ID NO: 2161
PIFPGG SEQ ID NO: 2162
IFPGGA SEQ ID NO: 2163


 


AAKFGA SEQ ID NO: 2164
AAKYGA SEQ ID NO: 2165
AAKAAK SEQ ID NO: 2166


 


AGLGAL SEQ ID NO: 2167
GAGVPG SEQ ID NO: 2002
GIPGGV SEQ ID NO: 2168


 


LKPVPG SEQ ID NO: 1905
PGVGVG SEQ ID NO: 2169
IPPAAA SEQ ID NO: 2170


 


ALVPGG SEQ ID NO: 2052
RPGVGV SEQ ID NO: 1938
ARPGFG SEQ ID NO: 2171


 


VLPGAR SEQ ID NO: 2172
GGFPGF SEQ ID NO: 2173
GLSPIF SEQ ID NO: 2174


 


PTGAGV SEQ ID NO: 2175
VGGVPG SEQ ID NO: 1941
GIPVVP SEQ ID NO: 1934


 


AGAAGK SEQ ID NO: 2176









Vimentin

Several candidate proteases may be responsible for the digestion of vimentin in fibrotic tissue We have through a range of in vitro cleavages of pure native proteins determined that the enzymes listed in the following table cleaved vimentin at least at the cleavage sites at each end of the following sequences or at the cleavage sites marked ‘.’ or where no ‘.’ is shown, at the ends of the sequences:







TABLE 27







Vimentin fragments generated by


specific proteases.











Aminoacid



Sequence between
residue


Protease
cleavage sites
numbers*





MMP2, MMP8,
RLRSSVPGVR. SEQ ID NO: 2177
69-78


Trypsin


 


MMP2, MMP8,
RLRSSVPGVL. SEQ ID NO: 2178
69-78


Trypsin


 


MMP2, MMP8,
.LLQDSVDFSL SEQ ID NO: 2179
79-89


Trypsin


 


MMP2, MMP8,
.FADLSEAANR SEQ ID NO: 2180
295-304


Trypsin


 


MMP2
.ISLPLPTFSS SEQ ID NO: 2181
410-420





*in the human vimentin sequence






Accordingly, in a method of the invention, said peptide fragments preferably comprise a neo-epitope formed by cleavage of vimentin by a protease at an N- or C-terminal site, or where indicated a site marked by the sign in any one of the partial sequences of vimentin in Table 24. The immunological binding partner may be one specifically reactive with a C-terminal or N-terminal neo-epitope formed by cleavage of vimentin. Suitable immunological binding partners may therefore be specifically reactive with any of the following sequences at the N terminal of a peptide:







TABLE 28





N-terminal sequences of protease generated


peptide fragments of vimentin.







Vimentin









LLQDSV
FADLSE
ISLPLP


SEQ ID NO: 2182
SEQ ID NO: 2183
SEQ ID NO: 2184









or with any of the following sequences at the C-terminal of a peptide:








TABLE 29





C-terminal sequences of protease


generated peptide fragments of vimentin.


Vimentin


















SVPGVR SEQ ID NO: 2185
SVPGVL SEQ ID NO: 2186









Further cleavage sites defining neo-epitopes that may be assayed in a similar manner can be identified by exposing collagens, elastin, CRP and proteoglycans or other fibrotic tissue proteins to any of the enzymes described herein and isolating and sequencing peptides thereby produced. Furthermore, assays may be based on the neo-epitopes generated adjacent the illustrated cleavage sites, i.e. in the C-terminal sequences that lead up to the N-terminal epitopes given above and the N-terminal sequences that connect to the C-terminal epitopes described.
Assays for more than one of the peptides described above may be conducted separately and their results combined or more than one of the peptides described above may be measured together.
The result of an assay according to the invention may be combined with one or more other measured biomarkers to form a composite index of diagnostic or prognostic value.
Generally, all previously known immunoassay formats can be used in accordance with this invention including heterogeneous and homogeneous formats, sandwich assays, competition assays, enzyme linked assays, radio-immune assays and the like. Thus, optionally, said method is conducted as a competition immunoassay in which said immunological binding partner and a competition agent are incubated in the presence of said sample and the competition agent competes with the peptide fragments in the sample to bind to the immunological binding partner.
Said competition agent may be (1) a synthetic peptide derived from the sequence of collagen type I, III, IV, V, or VI, or from CRP, or from any of the proteoglycans versican, lumican, perlecan, decorin and biglycan peptide, or a competition agent derived from (2) a purified native collagen type I, III, IV, V, or VI, or CRP, or any of the proteoglycans neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, perlecan, decorin and biglycan cleaved by proteases to reveal said neo-epitope.
One suitable method could be a competition immunoassay using monoclonal antibodies or antibody binding fragments binding to neo-epitopes of collagen type I, III, IV, V, VI, CRP, vimentin, or any of the proteoglycans neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments or neo-epitopes on peptide fragments from other proteins derived from fibrotic tissue. Appropriately selected synthetic peptides coated onto the solid surface of a microtitre plate could compete with the sample for binding to the monoclonal antibodies or binding fragments. Alternatively, purified, native collagen type I, III, IV, V, VI, CRP, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments carrying the neo-epitope recognised by the monoclonal antibody or binding fragment could be used on the solid surface. Yet another alternative is to immobilise the monoclonal antibody or binding fragment on the solid surface and then co-incubate the sample with a synthetic peptide appropriately linked to a signal molecule, e.g. horseradish peroxidase or biotin.
The sample may be a sample of serum, blood, plasma or other, e.g. fibrotic tissue biopsy.
Assays may be conducted as sandwich assays using a first immunological binding partner specifically reactive with a said neo-epitope and a second immunological binding partner reactive with the relevant protein to which the neo-epitope belongs. Optionally, said second immunological binding partner is directed to a second neo-epitope of the same protein.
In certain preferred methods the method further comprises comparing the determined level of said binding of said peptide fragments with values characteristic of (a) comparable healthy individuals and/or (b) a pathological fibrotic condition and optionally associating a higher level of the measured peptide (normally indicated by a higher level of binding) with a more severe degree of a said condition.
An aspect of the present invention relates to the development of monoclonal antibodies recognising neo-epitopes as described above, especially for collagen types I and IV. This can be achieved by immunising mice with synthetic peptides originating from the amino acid sequence of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan molecules (including the sequences listed above or sequences terminating therein), fusing the spleen-cells from selected mice to myeloma cells, and testing the monoclonal antibodies for binding to neo-epitopes on relevant synthetic peptides. Specificity for neo-epitopes can be ensured by requiring reactivity with a synthetic peptide and a lack of reactivity with either a C-prolongated form of the immunising peptide (for a C-terminal neo-epitope) or an N-terminal prolongated form of the immunising peptide (for an N-terminal neo-epitope). Antibodies for neo-epitopes may also be evaluated to establish a lack of binding capacity to native collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, pelecan and biglycan. Alternatively, specificity for a neo-epitope can be ensured by requiring the reactivity of the antibody to be negatively dependent on the presence of biotin or other functional groups covalently linked to one of the terminal amino acids.
The invention includes an immunological binding partner which is specifically immunoreactive with a neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan by a protease at a end-site in any one of the partial sequences of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan set out above, and may be for instance a monoclonal antibody or a binding fragment thereof.
The invention includes a cell line producing a monoclonal antibody against a C-terminal or N-terminal neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan at the end-sites of sequences in any one of the partial sequences of collagen type I, III, IV, V, VI, CRP, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan set out above.
The invention further provides a peptide comprising a C-terminal or N-terminal neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan in any one of the partial sequences of these proteins set out above. Such a peptide may be conjugated as a hapten to a carrier for producing an immune response to said peptide, or immobilised to a solid surface or conjugated to a detectable marker for use in an immunoassay.
The invention further comprises an isolated nucleic acid molecule coding for a peptide comprising a C-terminal or N-terminal neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan in any one of the partial sequences of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan set out above.
The invention further comprises a vector comprising a nucleic acid sequence comprising an expression signal and a coding sequence which codes for the expression of a peptide comprising a C-terminal or N-terminal neo-epitope formed by cleavage of collagen type I, III, IV, V, VI, CRP, vimentin, versican, lumican, decorin, perlecan and biglycan in any one of the partial sequences of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan set out above and further includes a host cell transformed with such a vector and expressing a said peptide.
Yet another aspect of the invention relates to kits, which can be used conveniently for carrying out the methods described above. Such kits may include (1) a microtitre plate coated with synthetic peptide; (2) a monoclonal antibody or antibody binding fragment of the invention reactive with said synthetic peptide; and (3) a labelled anti-mouse IgG immunoglobulin. Alternatively, such kits may include (1) a microtitre plate coated with purified native collagen type I, III, IV, V, VI, CRP, vimentin, versican, lumican, decorin, perlecan and biglycan fragments; (2) a monoclonal antibody recognising a neo-epitope on collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments and reactive with said purified collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan, and biglycan fragments; and (3) a labelled anti-mouse IgG immunoglobulin. Alternatively, such kits may include (1) a microtitre plate coated with streptavidin; (2) a synthetic peptide linked to biotin; (3) a monoclonal antibody recognising a neo-epitope on collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments and reactive with said synthetic peptide; and (4) a labelled anti-mouse IgG immunoglobulin. Yet another alternative could be kits including (1) a microtitre plate coated with streptavidin; (2) a synthetic peptide linked to biotin; (3) a monoclonal antibody recognising a neo-epitope on collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan fragments (and reactive with said synthetic peptide) and conjugated to horseradish peroxidase.
Thus, the invention includes an immunoassay kit comprising an immunological binding partner as described herein, especially in respect of collagens types I and IV, and a competition agent which binds said immunological binding partner, and optionally one or more of a wash reagent, a buffer, a stopping reagent, an enzyme label, an enzyme label substrate, calibration standards, an anti-mouse antibody and instructions.
The assays described herein are useful in the diagnosis of fibrosis in patients. In addition, the tests are useful for the assessment of disease progression, and the monitoring of response to therapy. The immunological binding partners of the invention may also be used in immunostaining to show the presence or location of collagen type I, III, IV, V, VI, CRP, vimentin, neurocan, brevican, fibromodulin, serglycins, syndecan, betaglycan, versican, lumican, decorin, perlecan and biglycan cleavage products.
EXEMPLIFICATION
EXAMPLE 1
Collagen Type III Degraded with MMP-9
Method
Cleavage: Collagen type III isolated from human placenta was dissolved in 10 mM acetic acid (1 mg/ml). The protein solution was then passed through a filter (Microcon Ultracel YM-10) to remove fragment contaminations. MMP-9 was preactivated with 4-aminophenylmercuric acetate (APMA, Sigma) at 37° C. for 3 hours. After activations, collagen type III and MMP-9 were mixed 100:1 and incubated shaking for 3 days at 37° C.
The solution was analyzed by liquid chromatography/mass spectrometry (LC/MS) and the fragments were identified by performing Mascot Search. The peptide sequences were selected by homology search, ensuring no cross-reactivity to other or related proteins, as well as interspecies cross-reactivity.
Antibody Design: The peptide sequences were synthesized and conjugated to ovalbumin (OVA). Mice were immunized ever 2-3 weeks, up to five. Antibody titers were checked by screening peptides, both selection and de-selection. When sufficient antibody titers were achieved, positive mice were selected for fusion, euthanized, and the spleen was disintegrated and B-cells were removed for fusion with myeloma cells. Selections of antibody producing cells were done by culturing and re-seeding the surviving chimera cells in single cell clones. Clones are selected by selection and de-selection peptides followed by native reactivity testing (FIG. 1), as neoepitopes are generated by synthetic small peptide sequences, which may not reflect the native proteins. An IgG subtype clone is selected for antibody production. Antibody purification is done by protein-G column.
Assay Development: Optimal antibody concentrations are determined by checker-board analysis, with dilutions of antibody coating and screening peptide, in competitions ELISA. The different determination for the collagen degraded by MMP-9 (CO3) assay is shown in Table 30. Table 30. Limit of Detection, Average Inter-and Intraassay variation of the CO3 assay.







TABLE 30





Limit of Detection, Average Inter- and Intraassay variation 


of the CO3 assay.


















Limit of Detection
0.5 ng/ml



Average Interassay variation
3.71%



Average Intraassay variation
5.48%










EXAMPLE 2
CO3 in Biological Relevant Samples
CO3 Levels in Bile Duct Ligated Rats Compared to Sham Operated Rats
Method: Forty female Sprague-Dawley rats (6 months old) were housed at the animal research facilities at Nordic Bioscience. The experiments were approved by the Experimental Animal Committee of the Danish Ministry of Justice, and were performed according to the European Standard for Good Clinical Practice (2008/561-1450). The rats were housed in standard type III-H cages at 18-22° C. with bedding and nest material (Altromin 1324; Altromin, Lage, Germany) and purified water (Milli-Q system; Millipore, Glostrup, Denmark) ad libitum. Rats were kept under conditions of a 12-hour light/dark cycle.
Liver fibrosis was induced by common BDL. In short: The rat was anaesthetized, the bile duct found, two ligations were performed around the bile duct followed by dissection between the ligations, the abdomen was closed. In sham operated rats, the abdomen was closed without bile duct ligation. The rats were divided into 2 groups: Group 1(10 BDL and 10 sham operated rats) were sacrificed after 2 weeks, and Group 2 (9 BDL and 10 sham operated rats) were sacrificed after 4 weeks. On completion of the study period (2, or 4 weeks), after at least 14 hours fasting, all surviving animals were asphyxiated by CO2 and sacrificed by exsanguinations.
Blood samples were taken from the retro-orbital sinus of at least 14 hours fasting rats under light CO2/O2 anaesthesia at baseline and at termination. The blood were collected and left 30 minutes at room temperature to cloth, followed by centrifugation at 1500 g for 10 minutes. All clot-free liquid were transferred to new tubes and centrifuged again at 1500 g for 10 minutes. The serum were then transferred to clean tubes and stored at −80° C.
CO3 were measured in ×5 diluted serum samples from the rats. Sham and BDL levels were compared by Mann-Whitneys two-tailed nonparametric test (α=0.05) of statistical significance assuming normal distribution.
CO3 levels increased significantly in the BDL groups compared to the Sham-operated animals. The results are shown in FIGS. 2A and 2B.
EXAMPLE 3
CO3 in Different Fibrotic Diseases (Human Serum)
CO3 levels were measured in serum from human with three different fibrotic diseases: Chronic obstructed pulmonary disease (COPD), Scleroderma, and Hepatitis virus C (HCV). The serum samples were retrieved from Sera Laboratories International Ltd (SLI Ltd), UK. CO3 levels were increased in the three different fibrotic diseases (FIG. 3).
EXAMPLE 4
Antibody Development—Detection of Marker CO3-610C
Type III collagen (Abcam, Cambridge, UK) was degraded in vitro by activated MMP-9 (Merck KGaA, Darmstadt, Germany) for 2 days. Degradation fragments were sequenced by LS-MS/MS and identified by MASCOT search. A specific peptide sequence 610KNGETGPQ (SEQ ID NO: 2251) was selected for antibody production. The N-terminal of this sequence is residue 610 of human collagen type III. The synthetic peptide was conjugated to ovalbumin prior to subcutaneous immunization of 4-6 week old Balb/C mice with about 200 μL emulsified antigen and 50 μg CO3-610C (KNGETGPQGPGGC-OVA), KNGETGPQGPGGC being SEQ ID NO: 2252. Consecutive immunizations were performed at two week intervals until stable sera titer levels were reached in Freund's incomplete adjuvant. The mice were bled from the second immunization on. At each bleeding, the serum titer was measured and the mouse with highest anti-serum titer was selected for fusion. After the fourth immunization, this mouse was rested for one month and then boosted intravenously with 50 μg CO3-610C in 100 μL 0.9% sodium chloride solution three days before isolation of the spleen for cell fusion.
Monoclonal antibody producing clones were selected using a) immunogenic peptide: KNGETGPQGP-GGC-Ovalbumine (OVA) (807678), b) screening peptide KNGETGPQGP-PG-K-Biotin (807971), KNGETGPQGP-PG-K being SEQ ID NO: 2253 c) de-selection peptides KDGETGAAGPPGK-Biotin (118318) representing a type II collagen alpha 1 chain (SEQ ID NO: 2254), KDGEAGAQGPPGK-Biotin representing a type I collagen alpha 1 chain degradation product, purchased from the Chinese Peptide Company, Beijing, China (SEQ ID NO: 2255). The ELISA coat plate was obtained from NUNC (Thermofisher, Copenhagen, Denmark). Peptide conjugation reagents and buffers were produced by Pierce (Thermofisher, Copenhagen, Denmark).
Buffer used for dissolving the coating peptide was composed of the following: 40 mM Na2HPO4, 12 H2O, 7 mM KH2PO4, 137 mM NaCl, 2.7 mM KCl, 25 mM EDTA, 0.1% Tween 20, 1% BSA, 10% sorbitol, pH 7. For a serum assay, buffer containing the following chemicals was used: 8 mM Na2HPO4, 12 H2O, 1.5 mM KH2PO4, 13.7 mM NaCl, 2.7 mM KCl, 0.1% Tween 20, 1% BSA, 0.003% phenol red, pH 7.4. A different buffer used for a urine assay contained 400 mM TRIZMA, 0.05% Tween 20, 0.1% BSA, 0.36% Bronidox L5, pH 8.0. For both serum and urine assays we used a washing buffer composed of 25 mM TRIZMA, 50 mM NaCl, 0.036% Bronidox L5, 0.1% Tween 20, and reaction-stopping buffer composed of 0.1% H2SO4. ELISA-plates used for the assay development were Streptavidin-coated from Roche (Hvidovre, Denmark) cat.: 11940279. All ELISA plates were analyzed with the ELISA reader from Molecular Devices, SpectraMax M, (CA. USA).
In preliminary experiments, we optimized the reagents, their concentrations and the incubation periods by performing several checkerboard analyses. A 96-well ELISA plate coated with streptavidin was further coated with 5 ng/ml of the synthetic peptide KNGETGPQGP-Biotinylated dissolved in PBS-TBE buffer at 20° C. for 30 minutes by constant shaking at 300 rpm (KNGETGPQGP being SEQ ID NO: 2256). After washing with washing buffer, 20 μl of sample was added, followed by 100 μl of peroxidase conjugated anti-human mAb-NB51-32 CO3-610C solution (23 pg/ml in incubation buffer). The plate was incubated for 1 hour at 20° C. during which time it was shaken at 300 rpm. This was followed by washing and finally, 100 μl tetramethylbenzinidine (TMB) (Kem-En-Tec cat.4380H) was dispensed and the plate incubated for 15 minutes in darkness and shaken at 300 rpm. In order to cease the reaction, 100 μl of stopping solution was added and the plate analyzed in the ELISA reader at 450 nm with 650 nm as reference.
A standard curve was performed by serial dilution of biotinylated-NB51-32 CO3-610C for a serum assay, and biotinylated-NB51-134 CO3-610C for a urine assay. Standard concentrations were 0, 0.33, 1, 3, 9, 27, 81 and 162 ng/ml. We designate fragments detected using the immunoassays so obtained as CO3-610C as the amino acid K at the N-terminal of the sequence KNGETGPQGP is amino acid 610 of the human collagen III sequence.
EXAMPLE 5
Comparison of CO3-610C and Other Biomarkers in Induced Liver Fibrosis in Rats
Animals
40 female Sprague-Dawley rats aged 6 months were housed at the animal research facilities at Nordic Bioscience, Copenhagen, Denmark. The experiments were approved by the Experimental Animal Committee of the Danish Ministry of Justice and were performed according to the European Standard for Good Clinical Practice (2008/561-1450). The rats were housed in standard type III-H cages at 18-22° C. with bedding and nest material (Altromin 1324; Altromin, Lage, Germany) and water ad libitum. Rats were kept under conditions of a 12-hour light/dark cycle.
Study Design
In 20 rats, liver fibrosis was induced by common BDL. The surgical procedure was performed under sterile conditions. The rat was anaesthetized, the bile duct localized and ligated in two places followed by dissection between the ligations, and the abdomen was closed. The other 20 rats were subjected to a sham operation, in which the abdomen was closed without bile duct ligation. The rats were then divided into 2 groups: Group 1 (10 BDL rats and 10 sham-operated rats) was sacrificed after 2 weeks and Group 2 (10 BDL and 10 sham-operated rats) was sacrificed after 4 weeks. On completion of the study period (2 or 4 weeks), after at least 14 hours fasting, all surviving animals were asphyxiated by CO2 and sacrificed by exsanguinations.
Blood Sampling
Blood samples were taken from the retro-orbital sinus of rats after at least 14 hours fasting, under light CO2/O2 anaesthesia, at baseline and at termination. Blood was left 30 minutes at room temperature to clot, followed by centrifugation at 1500 g for 10 minutes. All clot-free liquid was transferred to fresh tubes and centrifuged again at 1500 g for 10 minutes. The serum was then transferred to clean tubes and stored at −80° C.
Tissue Handling
After the rats were put down, their livers were carefully dissected, weighed, fixed in 4% formaldehyde for a minimum of 24 hours, cut into appropriate slices and embedded in paraffin. Sections 5 μm thick were cut, mounted on glass slides and stained with Sirius Red. The liver sections were evaluated histologically by assessment of the architecture, presence of inflammation, proliferation of bile ducts and fibrosis. The de novo bile duct formation in the parenchyma was evaluated semi-quantitatively using the following scoring system: normal=0, mild changes (1⁄3 or less of the lobule affected)=1, moderate changes (between 1⁄3 and 2⁄3 of the lobule affected)=2, and severe changes (2⁄3 or more of the lobule affected)=3. Digital photographs were captured using an Olympus BX60 microscope with ×40 and ×100 magnification and an Olympus 5050-zoom digital camera (Olympus, Tokyo, Japan).
Determination of Total Collagen and Serum CTX-II
The total collagen concentration was assayed using the commercial QuickZyme Collagen Assay (QuickZyme Bioscience, Leiden, The Netherlands). The concentration of CTX-II was assayed using the commercial Rat CTX-II kit (IDS Nordic, Herlev, Denmark). All samples were assayed in duplicate.
Type III Collagen mRNA Quantification
The number of transcripts of type III collagen (Col3a1) in liver tissue samples was determined by quantitative real-time polymerase chain reaction (RT-PCR) using fluorescent reporter probes. The number of Col3a1 copies in the sample was extrapolated from a standard curve obtained using Col3a1 plasmid cDNA Image Clone 7097081 (Geneservice, Cambridge, UK) as dilution standard. Amounts of Col3a1 were normalized with those of housekeeping gene hypoxanthine phosphoribosyltransferase 1 (Hprt1). Primers and probes for Col3a1 and Hprt1 mRNAs were designed using NCBI Reference Sequences NM_032085.1 and NM_012583.2 as templates, respectively (TIB Molbiol GmbH, Berlin, Germany). Total RNA was extracted from frozen liver samples using Absolutely RNA Miniprep kit (Stratagene, La Jolla, Calif., USA) following the manufacturer's instructions and its quality assessed in RNA Nano chips using a 2100 Bioanalyzer instrument (Agilent Technologies, Santa Clara, Calif., USA). Immediately after RNA isolation, complementary DNA (cDNA) was synthesised with Transcriptor First Strand cDNA Synthesis kit (Roche, Basel, Switzerland) using 1 μg of RNA as the template. For each sample tested, a cDNA synthesis negative control, omitting reverse transcriptase enzyme from the reaction mix, was included. Separate PCR reactions for Col3a1 and Hprt1 were performed in a 20 μL format using the Lightcycler Faststart DNA Master Plus Hybprobe kit (Roche) according to the manufacturer's instructions. Real time fluorescence data was collected in a Lightcycler 2.0 instrument (Roche).
Extractions
Tissue was pulverized in excess of liquid nitrogen in a steel mortar. Samples were then transferred into a 1.5 ml eppendorf tube and left shaking overnight at 4° C. in 0.5M Acetic Acid solution containing protease inhibitor cocktail (Roche). The samples were then sonicated with ultrasounds using 5 pulses at 60% amplitude (U50 control, IKA Labortechnik) and left for an additional 2 hours at 4° C. after which they were centrifuged for 5 minutes at 13,000 rpm. Supernatant was carefully removed, transferred in a new eppendorf and stored at −80° C.
Densitometry
Densitometry measurements were performed using UN-SCAN-IT Version 6.1 from Silk Scientific (give city, country).
Histology Image Analysis
Histology sections stained with Sirius Red were analyzed using Visiopharm software Version 3.2.8.0 (give city, country). Images were acquired using Pixelink PL-A623C microscope digital camera.
SDS PAGE and Western Blots
20 μg of tissue extract was mixed with loading buffer (Invitrogen LDS 4×, NP0007) containing the reducing agent (NuPAGE, NP0004 from Invitrogen). Samples were then loaded into 4-12% Bis-Tris gradient gel (NP0332BOX from Invitrogen) and run for 52 minutes at 200V. Proteins were then transferred onto a nitrocellulose membrane using the i-Blot transfer system (Invitrogen), blocked with 5% milk in TTBS overnight at 4 degrees. Beta Actin antibody (AbCam ab8229, give company, city country?) was used as a loading control.
Statistical Analysis
Mean values and standard error of the mean (SEM) were calculated using GraphPad Prism (GraphPad Software, Inc., San Diego, Calif., USA) and compared by Student's two-tailed paired t-test (α=0.05) of statistical significance assuming normal distribution, or by Mann-Whitney two-tailed nonparametric test (α=0.05). The coefficient of correlation (R2) and the corresponding p-value was determined by linear regression.
Results
Liver Appearance: At the time of sacrifice, livers of control animals showed normal gross morphology while livers of BDL animals were enlarged. The liver weights were significantly increased in BDL rats compared to the sham-operated controls (mean weights at sacrifice: 2 weeks post-surgery, sham 8.1 g; BDL 14.1 g; 4 weeks post-surgery, sham 9.0 g; BDL 19.4 g) (FIG. 4A). Semi-quantitative scoring of liver sections using the 0-3 scale showed significantly more structural changes of the liver at 4 weeks compared with 2 weeks (FIG. 4B). FIG. 4A shows liver weight in bile duct ligation (BDL)- or sham-operated rats. Data are shown as mean+SEM.[***, P<0.0001. FIG. 4B shows scoring of the structural changes in the liver of each group. Data are shown as mean+SEM. **, P=0.0094. Panel C shows Sirius Red photomicrographs showing the hepatic structure in sham-operated rats, and in BDL rats 2 and 4 weeks post-surgery. The hepatic structure around the portal tract is clearly disrupted in BDL rats compared with the sham-operated rats. Collagens are highlighted in red. Original magnification was ×40.
Under histological examination, the livers of sham-operated animals showed no sign of fibrosis and were microscopically normal (FIG. 4C). In the BDL livers, a marked ductal proliferation was observed. In the 2-week post-surgery group, the proliferation was located around the portal tract while in the 4-week group the proliferation had spread (FIG. 4C). Collagen deposition was found around the ductular structures. Inflammation was minimal and confined to the portal tracts. No signs of cholestasis were seen, whether intracellular cholestasis, bile plugs, bile infarctions or hepatocytic rosette formation.
Changes in CO3-610C Levels: FIG. 5 shows in panel A MMP-9 mediated CO3 degradation serum levels in bile duct ligated (BDL)- or sham-operated rats. Data are shown as mean+standard error of mean. 2 weeks post-surgery *** P<0.0001 and 4 weeks post-surgery ** P=0.0014. In panel B are shown CO3-610C delta values (termination-baseline paired), 2 weeks post-surgery P<0.0001 and 4 weeks post-surgery P=0.0016. In panel C are shown CTX-II levels in BDL- or sham-operated rats. Data are shown as mean+standard error of mean.
In the BDL groups CO3-610C levels increased significantly compared to sham groups (mean values: 2 weeks, post-surgery sham 39.7 ng/ml, BDL 100.3 ng/ml; average increase between the groups was 153%; 4 weeks post-surgery, sham 39.7, BDL 92.6 ng/ml, average increase between the groups was 133%) (FIGS. 5A-5B). There were no changes in the sham groups. CTX-II levels indicating collagen type II degradation did not change in the sham or BDL groups (FIG. 5C).
Type III Collagen Gene Expression: FIG. 6 shows Type III collagen gene expression in BDL or sham-operated rats. Data are shown as mean+standard of mean; 2 weeks post-surgery P<0.0001 and 4 weeks post-surgery P=0.0006
Type III collagen al chain mRNA increased significantly in both BDL groups compared with sham-operated rats.
Western Blot and Densitometry: FIG. 7 shows changes in the expression of CO3-610C in the liver of rats in BDL- and sham-operated groups assessed by A) Western blot 2 and 4 weeks post-surgery and B) Bands from western blot quantified by densitometry.
Western blot analysis showed very low levels of CO3-610C in sham-operated rats (FIG. 7A). At and after 2 weeks post-surgery CO3-610C levels prominently increased (FIG. 7A). Results were quantified by densitometry analysis (FIG. 7B).
Histology Image Analysis: FIG. 8A shows in the top row histology sections from BDL- or sham-operated rats stained with Sirius Red. The bottom row shows masked histology sections for quantifying total collagen content (red colour) in the liver. FIG. 8B shows total collagen quantified by Visiopharm software—2 weeks post-surgery P=0.0081; 4 weeks post-surgery P=0.0047. Histology sections stained with Sirius Red and enhanced using Visiopharm software showed increasing collagen content over time in BDL-operated rats. (FIG. 8A). The red color in the mask representing collagen was quantified using the same software (FIG. 8B) and confirmed a significant increase in total collagen content in BDL-operated rats compared with sham-operated rats (2 weeks post-surgery P=0.0081; 4 weeks post-surgery P=0.0047).
Correlation: FIG. 9A shows a correlation of Col3a1 to CO3-610C was found with R2=0.6993, P<0.0001. In FIG. 9B, a correlation of CO3-610C to % collagen was found with R2=0.2278 and P=0.0050. In panel C a correlation of Co13a1 to % collagen was found with R2=0.5409, P<0.0001. Correlations were found of the following: Col3a1 mRNA to CO3-610C with R2=0.6993 and P<0.0001 (FIG. 9A), and CO3-610C to % collagen quantified by visiopharm with R2=0.2278 and P=0.0050 (FIG. 9B), and Col3a1 mRNA to % collagen quantified by visiopharm with R2=0.5409 and P<0.0001 (FIG. 9C).
ECM remodelling is an integrated process of tissue development, maintenance and pathogenesis. Proteolytic activity is essential in this process for cell migration, removal of damaged tissue, and sequestering of new proteins, for the correct and optimal tissue orientation and quality (108:109). The specific matrix degradation products, neo-epitopes, may be important for the identification of new biochemical markers of liver fibrosis matrix turnover and understanding fibrosis pathogenesis. At present there are no available measuring techniques, nor biochemical markers, that allow for assessment of ECM remodeling in the pathogenesis of fibrosis.
In this example, to investigate the CO3-610C marker under in vivo situations, 6 months BDL rats were chosen, as they previously have been shown to have a lower collagen remodelling compared to younger rats. The rats are skeletally mature, and the growth plate is almost dormant, thereby contributing to a much lower extent to the overall collagen turnover. This influences the sensitivity and specificity for biomarker. These rats clearly presented with hepatic fibrosis, as evaluated by both quantitative histological analysis, and enlargement with increased weight, thus the model was an appropriate one to look for evidence of ECM remodeling, in particular for evidence of collagen type III in serum.
The present data clearly demonstrate the neo-epitope CO3-610C from MMP-9 mediated collagen type III degradation is a diagnostic biochemical marker for liver fibrosis with an average increases in serum of up to 153% from sham to BDL-operated rats.
To further investigate the biological rationale for the increased CO3-610C marker, we did protein extractions from healthy and diseased livers. By western blotting, we identified a predominant band, suggesting this to be an abundant protein fragment in diseased but not healthy livers. This provides evidence for the pathological accuracy of this novel marker.
To further investigate the pathological turnover representation of the liver, we measured type III collagen mRNA. We found an increase of mRNA in the BDL rats compared to those undergoing the sham operation, which correlates with previous findings. These data strongly suggest that liver fibrosis is not only an accumulation of ECM proteins, but also an accelerated turnover situation, in which both tissue formation and tissue degradation both are highly up regulated. Tissue formation outstrips tissue degradation, leading to an accumulation of scar tissue over time. Previous investigators have used other matrix turnover proteins to assess liver fibrosis, one being the type III collagen formation marker N-terminal type III pro-collagen. This marker represents collagen type III formation and has shown to be increased in liver fibrosis in previous studies.
To further understand and the dynamics of the biochemical makers CO3-610C, we did a range of correlations. Most importantly, there was a significant correlation of CO3-610C to the extent of fibrosis measured in the liver by quantitative histology. The level of liver fibrosis was correlated to the expression levels of the mRNA of collagen type III. Finally, the CO3-610C correlated to mRNA of collagen type III in the liver. Taken together, there was a significant correlation of the pathological processes in the liver with the levels of the systemic biochemical markers CO3-610C. In addition the tissue extractions provided evidence that the circulation levels were locally produced.
EXAMPLE 6
ELISA on Human Serum Samples
Human serum samples were obtained from patients with Chronic Obstructive Pulmonary Disease (COPD) (n=5), scleroderma (n=5), chronic hepatitis C virus infection (n=5), and healthy controls (n=5). The serum samples were tested in the CO3-610 ELISA (see Example 4 above) to determine the concentration of CO3-610 fragments. Results are shown in FIG. 10. While serum samples from the healthy subjects had concentration of CO3-610 fragments below 30 ng/ml, the diseased subjects were found to have elevated levels in circulation suggesting massive tissue remodelling in the affected fibrotic tissues.
EXAMPLE 7
Reactivity of Clone Nb94
Mice were immunized with synthetic peptide KAFVFP (SEQ ID NO: 1167) conjugated to ovalbumin (KAFVFPKESD-GGC-OVA (SEQ ID NO1049)), spleen cells were used for fusion, and monoclonal antibodies tested for reactivity to biotinylated KAFVFP (SEQ ID NO: 1167), i.e. (KAFVFPKESD-biotin (SEQ ID NO: 1049)) immobilized in wells of microtitre plates precoated with streptavidin. Antibodies binding to biotinylated KAFVFPKESD (SEQ ID NO: 1049), which could be inhibited by co-incubation with KAFVFPKESD (SEQ ID NO1049) but not the elongated peptide RKAFVFPKESD (SEQ ID NO: 1166), were selected for further characterization. The preferred monoclonal antibody was designated NB94-37-1A7.
Using a competition ELISA, essentially as described above with biotinylated KAFVFPKESD (SEQ ID NO: 1049) (used at 0.15 ng/ml) immobilized in the wells of streptavidin-coated microtitre plates, an incubation step (90 minutes at 20° C.) with sample and monoclonal antibody NB94-37-1A7 followed by a washing step, and then addition of peroxidase-conjugated anti-mouse immunoglobulins. For competition the following material was used in 2-fold dilutions; (1) the synthetic KAFVFP (SEQ ID NO: 1167) peptide; (2) a nonsense peptide (KNEGTG) unrelated to CRP; (3) a pool of human serum samples; (4) CRP proteolytically cleaved with MMP3 for 7 days, subsequently stopped by addition of EDTA to block protease activity, and stored at −80° C. until testing; (5) same as (4) but using MMP8 instead of MMP3; (6) same as (4) except using Cathepsin K (for 2 days) instead of MMP3 (and E64 as inhibitor to block Cathepsin K activity).
The data demonstrate that monoclonal antibody NB94-37-1A7 binds strongly to the synthetic peptide KAFVFPKESD (SEQ ID NO1049), and with CPR cleaved with MMP3 and MMP8. Cleavage of CRP with Cathepsin K release less analyte recognized by monoclonal antibody NB94-37-1A7. Finally, the data shows that the antibody binds to peptide fragments in human serum confirming the presence of this sequence in circulating peptide fragments.
EXAMPLE 8
CO3 in Biological Relevant Samples: CO3 Levels in Carbon Tetrachloride (CCl4)-Induced Cirrhosis in Rats
Animals and Induction of Cirrhosis
This study included 52 male Wistar rats with fibrosis or cirrhosis and 35 male Wistar control rats. To cause them to develop fibrosis or cirrhosis three-month old animals were included in an induction program with carbon tetrachloride (CCl4) and Phenobarbital treatment. CCl4 was administered by inhalation twice weekly and phenobarbital (0.3 g/l) added to the drinking water. Animals were allowed free access to water and food throughout the study.
Fibrosis Quantification
Liver sections (4 μm) were stained in 0.1% Sirius red F3B (Sigma-Aldrich, St. Louis, Mo.) in saturated picric acid (Sigma-Aldrich). Relative fibrosis area (expressed as a percentage of total liver area) was assessed by analyzing 36 fields of Sirius red-stained liver sections per animal. Each field was acquired at 10× magnification [E600 microscope (Nikon) and RT-Slider SPOT digital camera (Diagnostic Instruments, Inc., Sterling Heights, Mich.). Results were analyzed using a computerized Bioquant Life Science morphometry system. To evaluate the relative fibrosis area, the measured collagen area was divided by the net field area and then multiplied by 100. Subtraction of vascular luminal area from the total field area yielded the final calculation of the net fibrosis area. From each animal analyzed, the amount of fibrosis as percentage was measured and the average value presented.
Classification of Groups According to their Fibrosis/Cirrhosis Stage
Animals were classified into 4 different stages of fibrosis and cirrhosis (Group A: moderate fibrosis, group B: advanced fibrosis, Group C: moderate cirrhosis, and Group D: advanced cirrhosis) that were determined by the percentage of Sirius red positive liver area (Group A: <5%, Group B: 5 to 10%, Group C: 10 to 15% and Group D: >15%). For this purpose, control and fibrotic/cirrhotic rats were studied considering four different time points during the CCl4 treatment: 8, 12, 16 and 20 weeks after starting the cirrhosis induction program.
Hyaluronic Acid Measurement
Serum hyaluronan was measured using a sandwich ELISA kit (R&D Systems Inc., Minneapolis, Minn., USA).
Statistics
Statistical analysis of results was performed by unpaired Student's t tests when appropriate. Data were expressed as mean±S.E.M. and they were considered significant at a p level of 0.05 or less.
Study Design: Animals included in this protocol were randomly assigned to one of the following groups: A/eight weeks of CCl4 treatment, B/twelve weeks of CCl4 treatment, C/sixteen weeks of CCl4 treatment and D/twenty weeks of CCl4 treatment. In parallel, four control groups were studied at the same time points. Thirteen fibrotic rats and seven control rats were included in each group. At the end of the study, rats were placed in standard metabolic cages (Tecniplast Deutschland, Hohenpeissenberg, Germany) during an adaptation period of 3 days before proceeding with the twenty-four-hour urine collection. Urinary volumes were determined gravimetrically. During the adaptation period, rats were allowed to get free access to tap water and food. Then, 24-hour urine samples were centrifuged for 5 min at 2,500 rpm and aliquoted into ten polypropylene tubes (400 μL each). Urine samples were stored at −80° C. for subsequent analysis.
At scheduled necropsies, rats were weighed, anesthetized with pentobarbital (50 mg/kl) and decapitated. Blood were collected and allowed to stand at room temperature for 20 min to allow clotting and then centrifuged for 10 min at 2500 rpm. Serum were collected in polypropylene tubes aliquots (400 μl each) and transferred via dry ice to a −80° C. freezer. Collection of baseline blood samples at the beginning of the CCl4 treatment was not considered in order to avoid additional intervention that may increase the risk of infection and/or introduce modifications in the experimental model that may compromise the evolution of the induced pathophysiological process. For histology and Sirius red staining, half of the left lobe of the liver was placed in 10% neutral buffered formalin for 16 hours, embedded in paraffin and sectioned into 4-μm-thick slices. After liver fibrosis quantification, the unused paraffin block material was preserved for biomarker quantification. The other half of the left lobe was flash-frozen in liquid nitrogen and stored for Western blot, RT-PCR or immunohistochemical analysis. Measurements of liver fibrotic area, serum and urine osmolality, Na+ and K+, albumin, creatinine, alanine amino-transferase and lactate dehydrogenase were made according to the Material and Methods section.
Results
Histological Validation of the Model:
Liver collagen was quantified in all study animals by Sirius red staining of liver slices. The final data for each animal was taken as the average of red staining observed in 36 consecutive microscope fields (FIG. 12).
FIG. 12 shows representative pictures from two sets of 36 images used to quantify collagen accumulation in liver in rat #1 (left) and rat #43 (right) treated with carbon tetrachloride for eight and twenty weeks respectively.
The serum CO3 marker shows statistically significant increases in both fibrotic and cirrhotic rats compared to control rats. Animals were classified according to a fully automated syrius red staining of the liver procedure used to quantify fibrosis (FIGS. 13-14).
FIG. 13 shows serum CO3 levels in CCl4 inhalation and control rats as performed in Hospital Clinic (Barcelona). Each point represents one animal. Rats were classified according a computerized image analysis method of syrius red staining of the liver used to quantify fibrosis.
When quantitative values of serum CO3 and syrius red staining of the liver were studied in each individual animal, we found a statistically significant correlation between the two variables (R2=0.4087; n=21) (FIG. 14).
We have compared the levels of CO3-610C with the serological benchmark of liver fibrosis hyaluronic acid (HA). HA levels were quantified with a commercial ELISA kit and results show significant elevations of this ECM component in cirrhotic rats vs. fibrotic animals (FIGS. 15, 16A-16B). The correlation of CO3 to Sirius red outperformed that of HA. More than seventy percent of the variation in liver fibrosis histological quantification can be explained by the serological measurement of CO3. The remaining thirty percent is due to unknown variables or inherent variability. Instead only 25% of liver fibrosis can be explained by measuring hyaluronic acid (FIG. 15). As expected from the previous result no correlation could be found between CO3 and hyaluronic acid suggesting that they are the result of two independent pathophysiological processes in the development of liver fibrosis (FIG. 17).
EXAMPLE 9
Bleomycin Induced Skin Fibrosis in Mice
Mice were treated by application to the skin of PBS or bleomycin. Increasing levels in urine of the MMP-9 mediated collagen III (CO3) degradation fragment CO3-610 were associated with skin fibrosis progression in mice.
FIGS. 18A-18B shows a skin section from a PBS treated mouse at 8 weeks of treatment (FIG. 18A) and a skin section from Bleomycin treated mouse at 8 weeks of treatment (FIG. 18B). Skin thickness increase between PBS (n=7/time point) and Bleomycin (n=13/time point) treated mice for 2 weeks (P=0.0029), 4 weeks (P=0.0004), 6 weeks (P<0.0001) and 8 weeks (P<0.0001) is plotted in FIGS. 18C-18D. Overall skin thickness increase between PBS (n=28) and Bleomycin (n=52) treated mice for the duration of the study (P<0.0001). Skin width was calculated by Visiopharm software as an overall number per skin section instead of sampling pictures.
FIG. 19 shows CO3-610 urine assay results which demonstrate a significant increase throughout the time points of the study. The figure shows result per time point (n=7 PBS, n=13 Bleomycin treated per termination point) and collective CO3-610 levels for all time points (n=28 PBS and n=52 Bleomycin treated mice). 2 weeks P=0.0008, 4 weeks P<0.0001, 6 weeks P<0.0001, 8 weeks P<0.0001 and overall P<0.0001.
FIGS. 20A-20B shows a CO3-610 Western blots image with control C and Bleomycin B after 2 and 8 weeks treatment (FIG. 20A). CO3-610 densitometry measurements for all time points (n=7 PBS and n=13 Bleomycin treated per termination point) and collective CO3-610 levels (n=28 PBS and n=52 Bleomycin treated mice) are shown in FIG. 20B, demonstrating a statistically significant increase of CO3-610 levels (P<0.0001).
As seen in FIG. 21, CO3-610 levels in urine assay were found to be correlated with skin thickness progression, and therefore total collagen deposition r=0.4883, R2=0.2384.
As seen in FIG. 22, statistically significant correlation was found (r=0.6528, P<0.0001) between results from the CO3-610 ELISA urine assay and Western blot densitometry measurements.
In this specification, unless expressly otherwise indicated, the word ‘or’ is used in the sense of an operator that returns a true value when either or both of the stated conditions is met, as opposed to the operator ‘exclusive or’ which requires that only one of the conditions is met. The word ‘comprising’ is used in the sense of ‘including’ rather than in to mean ‘consisting of’. All prior teachings acknowledged above are hereby incorporated by reference. No acknowledgement of any prior published document herein should be taken to be an admission or representation that the teaching thereof was common general knowledge in Australia or elsewhere at the date hereof.
The following references are cited herein:
1. World Health Organization. Reducing Risks, Promoting Healthy Life. Reducing Risks, Promoting Healthy Life, Geneva: WHO, 2002:1-230.
2. Wynn T A. Cellular and molecular mechanisms of fibrosis. J Pathol 2008; 214:199-210.
3. Friedman S L. Mechanisms of disease: Mechanisms of hepatic fibrosis and therapeutic implications. Nat Clin Pract Gastroenterol Hepatol 2004; 1:98-105.
4. Tomasek J J, Gabbiani G, Hinz B, Chaponnier C, Brown R A. Myofibroblasts and mechano-regulation of connective tissue remodelling. Nat Rev Mol Cell Biol 2002; 3:349-363.
5. Wynn T A. Common and unique mechanisms regulate fibrosis in various fibroproliferative diseases. J Clin Invest 2007; 117:524-529.
6. Marcellin P, Asselah T, Boyer N. Fibrosis and disease progression in hepatitis C. Hepatology 2002; 36:S47-S56.
7. Gagliano N, Arosio B, Grizzi F, Masson S, Tagliabue J, Dioguardi N, Vergani C, Annoni G. Reduced collagenolytic activity of matrix metalloproteinases and development of liver fibrosis in the aging rat. Mech Ageing Dev 2002; 123:413-425.
8. Laurent G J. Dynamic state of collagen: pathways of collagen degradation in vivo and their possible role in regulation of collagen mass. Am J Physiol 1987; 252:C1-C9.
9. Mays P K, McAnulty R J, Campa J S, Laurent G J. Age-related changes in collagen synthesis and degradation in rat tissues. Importance of degradation of newly synthesized collagen in regulating collagen production. Biochem J 1991; 276 (Pt 2):307-313.
10. Garrone R, Lethias C, Le Guellec D. Distribution of minor collagens during skin development. Microsc Res Tech 1997; 38:407-412.
11. Gelse K, Poschl E, Aigner T. Collagens—structure, function, and biosynthesis. Adv Drug Deliv Rev 2003; 55:1531-1546.
12. Phan S H, Thrall R S. Pulmonary Fibrosis. Lung Biology in Health and Disease. 80 ed. New York: Marcel Dekker, Inc., 1995.
13. Martinez-Hernandez A, Amenta P S. The hepatic extracellular matrix. II. Ontogenesis, regeneration and cirrhosis. Virchows Arch A Pathol Anat Histopathol 1993; 423:77-84.
14. Gilliam A C. Scleroderma. Curr Dir Autoimmun 2008; 10:258-279.
15. Gressner A M, Weiskirchen R. Modern pathogenetic concepts of liver fibrosis suggest stellate cells and TGF-beta as major players and therapeutic targets. J Cell Mol Med 2006; 10:76-99.
16. Heinegard D, Oldberg A. Structure and biology of cartilage and bone matrix noncollagenous macromolecules. FASEB J 1989; 3:2042-2051.
17. Svensson L, Oldberg A, Heinegard D. Collagen binding proteins. Osteoarthritis and Cartilage 2001; 9:S23-S28.
18. Kiani C, Chen L, Wu Y J, Yee A J, Yang B B. Structure and function of aggrecan. Cell Res 2002; 12:19-32.
19. Krusius T, Gehlsen K R, Ruoslahti E. A fibroblast chondroitin sulfate proteoglycan core protein contains lectin-like and growth factor-like sequences. J Biol Chem 1987; 262:13120-13125.
20. Yang B L, Zhang Y, Cao L, Yang B B. Cell adhesion and proliferation mediated through the G1 domain of versican. J Cell Biochem 1999; 72:210-220.
21. Rauch U, Karthikeyan L, Maurel P, Margolis R U, Margolis R K. Cloning and primary structure of neurocan, a developmentally regulated, aggregating chondroitin sulfate proteoglycan of brain. J Biol Chem 1992; 267:19536-19547.
22. Yamada H, Watanabe K, Shimonaka M, Yamaguchi Y. Molecular cloning of brevican, a novel brain proteoglycan of the aggrecan/versican family. J Biol Chem 1994; 269:10119-10126.
23. Blochberger T C, Cornuet P K, Hassell J R. Isolation and partial characterization of lumican and decorin from adult chicken corneas. A keratan sulfate-containing isoform of decorin is developmentally regulated. J Biol Chem 1992; 267:20613-20619.
24. Fisher L W, Termine J D, Young M F. Deduced protein sequence of bone small proteoglycan I (biglycan) shows homology with proteoglycan II (decorin) and several nonconnective tissue proteins in a variety of species. J Biol Chem 1989; 264:4571-4576.
25. Toyama-Sorimachi N, Sorimachi H, Tobita Y, Kitamura F, Yagita H, Suzuki K, Miyasaka M. A novel ligand for CD44 is serglycin, a hematopoietic cell lineage-specific proteoglycan. Possible involvement in lymphoid cell adherence and activation. J Biol Chem 1995; 270:7437-7444.
26. Bartlett A H, Hayashida K, Park P W. Molecular and cellular mechanisms of syndecans in tissue injury and inflammation. Mol Cells 2007; 24:153-166.
27. Lopez-Casillas F, Wrana J L, Massague J. Betaglycan presents ligand to the TGF beta signaling receptor. Cell 1993; 73:1435-1444.
28. Olsen B R. Life without perlecan has its problems. J Cell Biol 1999; 147:909-912.
29. Gabay C, Kushner I. Acute-phase proteins and other systemic responses to inflammation. N Engl J Med 1999; 340:448-454.
30. Benyon R C, Arthur M J. Extracellular matrix degradation and the role of hepatic stellate cells. Semin Liver Dis 2001; 21:373-384.
31. Guo J, Friedman SL. Hepatic fibrogenesis. Semin Liver Dis 2007; 27:413-426.
32. Iredale J P, Benyon R C, Arthur M J, Ferris W F, Alcolado R, Winwood P J, Clark N, Murphy G. Tissue inhibitor of metalloproteinase-1 messenger RNA expression is enhanced relative to interstitial collagenase messenger RNA in experimental liver injury and fibrosis. Hepatology 1996; 24:176-184.
33. Lee K N, Jackson K W, Christiansen V J, Lee C S, Chun J G, McKee P A. Antiplasmin-cleaving enzyme is a soluble form of fibroblast activation protein. Blood 2006; 107:1397-1404.
34. Acharya P S, Zukas A, Chandan V, Katzenstein A L, Pure E. Fibroblast activation protein: a serine protease expressed at the remodeling interface in idiopathic pulmonary fibrosis. Hum Pathol 2006; 37:352-360.
35. Levy M T, McCaughan G W, Marinos G, Gorrell M D. Intrahepatic expression of the hepatic stellate cell marker fibroblast activation protein correlates with the degree of fibrosis in hepatitis C virus infection. Liver 2002; 22:93-101.
36. Meyer O. Prognostic markers for systemic sclerosis. Joint Bone Spine 2006; 73:490-494.
37. Hummers L K. Microvascular damage in systemic sclerosis: detection and monitoring with biomarkers. Curr Rheumatol Rep 2006; 8:131-137.
38. McHugh N J, Distler O, Giacomelli R, Riemekasten G. Non organ based laboratory markers in systemic sclerosis. Clin Exp Rheumatol 2003; 21:S32-S38.
39. Muller-Quernheim J. Serum markers for the staging of disease activity of sarcoidosis and other interstitial lung diseases of unknown etiology. Sarcoidosis Vasc Diffuse Lung Dis 1998; 15:22-37.
40. Gressner O A, Weiskirchen R, Gressner A M. Biomarkers of liver fibrosis: clinical translation of molecular pathogenesis or based on liver-dependent malfunction tests. Clin Chim Acta 2007; 381:107-113.
41. Gressner O A, Weiskirchen R, Gressner A M. Biomarkers of hepatic fibrosis, fibrogenesis and genetic pre-disposition pending between fiction and reality. J Cell Mol Med 2007; 11:1031-1051.
42. Mariat C. [Diagnosis and follow-up of chronic kidney graft dysfunction: from DFG to new biomarkers]. Nephrol Ther 2008; 4 Suppl 3:S204-S207.
43. Yoneda M, Mawatari H, Fujita K, Iida H, Yonemitsu K, Kato S, Takahashi H, Kirikoshi H, Inamori M, Nozaki Y, Abe Y, Kubota K, Saito S, Iwasaki T, Terauchi Y, Togo S, Maeyama S, Nakajima A. High-sensitivity C-reactive protein is an independent clinical feature of nonalcoholic steatohepatitis (NASH) and also of the severity of fibrosis in NASH. J Gastroenterol 2007; 42:573-582.
44. Wong V S, Hughes V, Trull A, Wight D G, Petrik J, Alexander G J. Serum hyaluronic acid is a useful marker of liver fibrosis in chronic hepatitis C virus infection. J Viral Hepat 1998; 5:187-192.
45. Parise E R, Oliveira A C, Figueiredo-Mendes C, Lanzoni V, Martins J, Nader H, Ferraz M L. Noninvasive serum markers in the diagnosis of structural liver damage in chronic hepatitis C virus infection. Liver Int 2006; 26:1095-1099.
46. McHutchison J G, Blatt L M, de Medina M, Craig J R, Conrad A, Schiff E R, Tong M J. Measurement of serum hyaluronic acid in patients with chronic hepatitis C and its relationship to liver histology. Consensus Interferon Study Group. J Gastroenterol Hepatol 2000; 15:945-951.
47. Camacho V R, Silveira T R, Oliveira J R, Barros S G, Cerski C T. Relationship between serum concentrations of type III procollagen, hyluronic acid and histopathological findings in the liver of HCV-positive blood donors. Arq Gastroenterol 2007; 44:118-122.
48. Lorenzo-Zuniga V, Bartoli R, Masnou H, Montoliu S, Morillas R M, Planas R. Serum concentrations of insulin-like growth factor-I (igf-I) as a marker of liver fibrosis in patients with chronic hepatitis C. Dig Dis Sci 2007; 52:3245-3250.
49. Manolakopoulos S, Bethanis S, Liapi C, Stripeli F, Sklavos P, Margeli A, Christidou A, Katsanika A, Vogiatzakis E, Tzourmakliotis D, Theocharis S. An assessment of serum leptin levels in patients with chronic viral hepatitis: a prospective study. BMC Gastroenterol 2007; 7:17.
50. Camacho V R, Silveira T R, Oliveira J R, Barros S G, Cerski C T. Relationship between serum concentrations of type III procollagen, hyluronic acid and histopathological findings in the liver of HCV-positive blood donors. Arq Gastroenterol 2007; 44:118-122.
51. Leroy V, Hilleret M N, Sturm N, Trocme C, Renversez J C, Faure P, Morel F, Zarski J P. Prospective comparison of six non-invasive scores for the diagnosis of liver fibrosis in chronic hepatitis C. J Hepatol 2007; 46:775-782.
52. Trocme C, Leroy V, Sturm N, Hilleret M N, Bottari S, Morel F, Zarski J P. Longitudinal evaluation of a fibrosis index combining MMP-1 and PIIINP compared with MMP-9, TIMP-1 and hyaluronic acid in patients with chronic hepatitis C treated by interferon-alpha and ribavirin. J Viral Hepat 2006; 13:643-651.
53. Zheng M, Cai W M, Weng H L, Liu R H. ROC curves in evaluation of serum fibrosis indices for hepatic fibrosis. World J Gastroenterol 2002; 8:1073-1076.
54. Lebensztejn D M, Sobaniec-Lotowska M E, Bauer M, Kaczmarski M, Voelker M, Schuppan D. Serum fibrosis markers as predictors of an antifibrotic effect of interferon alfa in children with chronic hepatitis B. Eur J Gastroenterol Hepatol 2005; 17:843-848.
55. Lebensztejn D M, Sobaniec-Lotowska M E, Kaczmarski M, Voelker M, Schuppan D. Matrix-derived serum markers in monitoring liver fibrosis in children with chronic hepatitis B treated with interferon alpha. World J Gastroenterol 2006; 12:3338-3343.
56. Tsochatzis E, Papatheodoridis G V, Hadziyannis E, Georgiou A, Kafiri G, Tiniakos D G, Manesis E K, Archimandritis A J. Serum adipokine levels in chronic liver diseases: association of resistin levels with fibrosis severity. Scand J Gastroenterol 2008; 43:1128-1136.
57. Patel K, Gordon S C, Jacobson I, Hezode C, Oh E, Smith K M, Pawlotsky J M, McHutchison J G. Evaluation of a panel of non-invasive serum markers to differentiate mild from moderate-to-advanced liver fibrosis in chronic hepatitis C patients. J Hepatol 2004; 41:935-942.
58. Lieber C S, Weiss D G, Paronetto F. Value of fibrosis markers for staging liver fibrosis in patients with precirrhotic alcoholic liver disease. Alcohol Clin Exp Res 2008; 32:1031-1039.
59. Forns X, Ampurdanes S, Llovet J M, Aponte J, Quinto L, Martinez-Bauer E, Bruguera M, Sanchez-Tapias J M, Rodes J. Identification of chronic hepatitis C patients without hepatic fibrosis by a simple predictive model. Hepatology 2002; 36:986-992.
60. Bourliere M, Penaranda G, Renou C, Botta-Fridlund D, Tran A, Portal I, Lecomte L, Castellani P, Rosenthal-Allieri M A, Gerolami R, Ouzan D, Deydier R, Degott C, Halfon P. Validation and comparison of indexes for fibrosis and cirrhosis prediction in chronic hepatitis C patients: proposal for a pragmatic approach classification without liver biopsies. J Viral Hepat 2006; 13:659-670.
61. Cacoub P, Carrat F, Bedossa P, Lambert J, Penaranda G, Perronne C, Pol S, Halfon P. Comparison of non-invasive liver fibrosis biomarkers in HIV/HCV co-infected patients: the fibrovic study—ANRS HC02. J Hepatol 2008; 48:765-773.
62. Nunes D, Fleming C, Offner G, O'Brien M, Tumilty S, Fix O, Heeren T, Koziel M, Graham C, Craven D E, Stuver S, Horsburgh C R, Jr. HIV infection does not affect the performance of noninvasive markers of fibrosis for the diagnosis of hepatitis C virus-related liver disease. J Acquir Immune Defic Syndr 2005; 40:538-544.
63. Grigorescu M, Rusu M, Neculoiu D, Radu C, Serban A, Catanas M, Grigorescu M D. The FibroTest value in discriminating between insignificant and significant fibrosis in chronic hepatitis C patients. The Romanian experience. J Gastrointestin Liver Dis 2007; 16:31-37.
64. Halfon P, Bacq Y, De M A, Penaranda G, Bourliere M, Ouzan D, Tran A, Botta D, Renou C, Brechot M C, Degott C, Paradis V. Comparison of test performance profile for blood tests of liver fibrosis in chronic hepatitis C. J Hepatol 2007; 46:395-402.
65. Halfon P, Bourliere M, Deydier R, Botta-Fridlund D, Renou C, Tran A, Portal I, Allemand I, Bertrand J J, Rosenthal-Allieri A, Rotily M, Sattonet C, Benderitter T, Saint Paul M C, Bonnot H P, Penaranda G, Degott C, Masseyeff M F, Ouzan D. Independent prospective multicenter validation of biochemical markers (fibrotest-actitest) for the prediction of liver fibrosis and activity in patients with chronic hepatitis C: the fibropaca study. Am J Gastroenterol 2006; 101:547-555.
66. Leroy V, Halfon P, Bacq Y, Boursier J, Rousselet M C, Bourliere M, De M A, Sturm N, Hunault G, Penaranda G, Brechot M C, Trocme C, Cales P. Diagnostic accuracy, reproducibility and robustness of fibrosis blood tests in chronic hepatitis C: a meta-analysis with individual data. Clin Biochem 2008; 41:1368-1376.
67. Ratziu V, Massard J, Charlotte F, Messous D, Imbert-Bismut F, Bonyhay L, Tahiri M, Munteanu M, Thabut D, Cadranel J F, Le B B, de L, V, Poynard T. Diagnostic value of biochemical markers (FibroTest-FibroSURE) for the prediction of liver fibrosis in patients with non-alcoholic fatty liver disease. BMC Gastroenterol 2006; 6:6.
68. Poynard T, Imbert-Bismut F, Ratziu V, Chevret S, Jardel C, Moussalli J, Messous D, Degos F. Biochemical markers of liver fibrosis in patients infected by hepatitis C virus: longitudinal validation in a randomized trial. J Viral Hepat 2002; 9:128-133.
69. Poynard T, Munteanu M, Imbert-Bismut F, Charlotte F, Thabut D, Le C S, Messous D, Thibault V, Benhamou Y, Moussalli J, Ratziu V. Prospective analysis of discordant results between biochemical markers and biopsy in patients with chronic hepatitis C. Clin Chem 2004; 50:1344-1355.
70. Poynard T, Morra R, Halfon P, Castera L, Ratziu V, Imbert-Bismut F, Naveau S, Thabut D, Lebrec D, Zoulim F, Bourliere M, Cacoub P, Messous D, Munteanu M, de L, V. Meta-analyses of FibroTest diagnostic value in chronic liver disease. BMC Gastroenterol 2007; 7:40.
71. Ngo Y, Munteanu M, Messous D, Charlotte F, Imbert-Bismut F, Thabut D, Lebray P, Thibault V, Benhamou Y, Moussalli J, Ratziu V, Poynard T. A prospective analysis of the prognostic value of biomarkers (FibroTest) in patients with chronic hepatitis C. Clin Chem 2006; 52:1887-1896.
72. Naveau S, Raynard B, Ratziu V, Abella A, Imbert-Bismut F, Messous D, Beuzen F, Capron F, Thabut D, Munteanu M, Chaput J C, Poynard T. Biomarkers for the prediction of liver fibrosis in patients with chronic alcoholic liver disease. Clin Gastroenterol Hepatol 2005; 3:167-174.
73. Myers R P, Tainturier M H, Ratziu V, Piton A, Thibault V, Imbert-Bismut F, Messous D, Charlotte F, Di M, V, Benhamou Y, Poynard T. Prediction of liver histological lesions with biochemical markers in patients with chronic hepatitis B. J Hepatol 2003; 39:222-230.
74. Jacqueminet S, Lebray P, Morra R, Munteanu M, Devers L, Messous D, Bernard M, Hartemann-Heurtier A, Imbert-Bismut F, Ratziu V, Grimaldi A, Poynard T. Screening for liver fibrosis by using a noninvasive biomarker in patients with diabetes. Clin Gastroenterol Hepatol 2008; 6:828-831.
75. Nunes D, Fleming C, Offner G, O'Brien M, Tumilty S, Fix O, Heeren T, Koziel M, Graham C, Craven D E, Stuver S, Horsburgh C R, Jr. HIV infection does not affect the performance of noninvasive markers of fibrosis for the diagnosis of hepatitis C virus-related liver disease. J Acquir Immune Defic Syndr 2005; 40:538-544.
76. Poynard T, Zoulim F, Ratziu V, Degos F, Imbert-Bismut F, Deny P, Landais P, El H A, Slama A, Blin P, Thibault V, Parvaz P, Munteanu M, Trepo C. Longitudinal assessment of histology surrogate markers (FibroTest-ActiTest) during lamivudine therapy in patients with chronic hepatitis B infection. Am J Gastroenterol 2005; 100:1970-1980.
77. Poynard T, Munteanu M, Imbert-Bismut F, Charlotte F, Thabut D, Le C S, Messous D, Thibault V, Benhamou Y, Moussalli J, Ratziu V. Prospective analysis of discordant results between biochemical markers and biopsy in patients with chronic hepatitis C. Clin Chem 2004; 50:1344-1355.
78. Myers R P, Tainturier M H, Ratziu V, Piton A, Thibault V, Imbert-Bismut F, Messous D, Charlotte F, Di M, V, Benhamou Y, Poynard T. Prediction of liver histological lesions with biochemical markers in patients with chronic hepatitis B. J Hepatol 2003; 39:222-230.
79. Carvalho-Filho R J, Schiavon L L, Narciso-Schiavon J L, Sampaio J P, Lanzoni V P, Ferraz M L, Silva A E. Optimized cutoffs improve performance of the aspartate aminotransferase to platelet ratio index for predicting significant liver fibrosis in human immunodeficiency virus/hepatitis C virus co-infection. Liver Int 2008; 28:486-493.
80. Al-Mohri H, Cooper C, Murphy T, Klein M B. Validation of a simple model for predicting liver fibrosis in HIV/hepatitis C virus-coinfected patients. HIV Med 2005; 6:375-378.
81. Cales P, Laine F, Boursier J, Deugnier Y, Moal V, Oberti F, Hunault G, Rousselet M C, Hubert I, Laafi J, Ducluzeaux P H, Lunel F. Comparison of blood tests for liver fibrosis specific or not to NAFLD. J Hepatol 2008.
82. Paggi S, Colli A, Fraquelli M, Vigano M, Del P P, Facciotto C, Colombo M, Ronchi G, Conte D. A non-invasive algorithm accurately predicts advanced fibrosis in hepatitis C: a comparison using histology with internal-external validation. J Hepatol 2008; 49:564-571.
83. Trang T, Petersen J R, Snyder N. Non-invasive markers of hepatic fibrosis in patients co-infected with HCV and HIV: comparison of the APRI and FIB-4 index. Clin Chim Acta 2008; 397:51-54.
84. Snyder N, Gajula L, Xiao S Y, Grady J, Luxon B, Lau D T, Soloway R, Petersen J. APRI: an easy and validated predictor of hepatic fibrosis in chronic hepatitis C. J Clin Gastroenterol 2006; 40:535-542.
85. Snyder N, Nguyen A, Gajula L, Soloway R, Xiao S Y, Lau D T, Petersen J. The APRI may be enhanced by the use of the FIBROSpect II in the estimation of fibrosis in chronic hepatitis C. Clin Chim Acta 2007; 381:119-123.
86. Hongbo L, Xiaohui L, Hong K, Wei W, Yong Z. Assessing routine and serum markers of liver fibrosis in CHB patients using parallel and serial interpretation. Clin Biochem 2007; 40:562-566.
87. Nunes D, Fleming C, Offner G, O'Brien M, Tumilty S, Fix O, Heeren T, Koziel M, Graham C, Craven D E, Stuver S, Horsburgh C R, Jr. HIV infection does not affect the performance of noninvasive markers of fibrosis for the diagnosis of hepatitis C virus-related liver disease. J Acquir Immune Defic Syndr 2005; 40:538-544.
88. Adams L A, Bulsara M, Rossi E, DeBoer B, Speers D, George J, Kench J, Farrell G, McCaughan G W, Jeffrey G P. Hepascore: an accurate validated predictor of liver fibrosis in chronic hepatitis C infection. Clin Chem 2005; 51:1867-1873.
89. Koda M, Matunaga Y, Kawakami M, Kishimoto Y, Suou T, Murawaki Y. Fibrolndex, a practical index for predicting significant fibrosis in patients with chronic hepatitis C. Hepatology 2007; 45:297-306.
90. Metwally M A, Zein C O, Zein N N. Predictors and noninvasive identification of severe liver fibrosis in patients with chronic hepatitis C. Dig Dis Sci 2007; 52:582-588.
91. Mohamadnejad M, Montazeri G, Fazlollahi A, Zamani F, Nasiri J, Nobakht H, Forouzanfar M H, Abedian S, Tavangar S M, Mohamadkhani A, Ghoujeghi F, Estakhri A, Nouri N, Farzadi Z, Najjari A, Malekzadeh R. Noninvasive markers of liver fibrosis and inflammation in chronic hepatitis B-virus related liver disease. Am J Gastroenterol 2006; 101:2537-2545.
92. Zaman A, Rosen H R, Ingram K, Corless C L, Oh E, Smith K. Assessment of FIBROSpect II to detect hepatic fibrosis in chronic hepatitis C patients. Am J Med 2007; 120:280-14.
93. Patel K, Nelson D R, Rockey D C, Afdhal N H, Smith K M, Oh E, Hettinger K, Vallee M, Dev A, Smith-Riggs M, McHutchison J G. Correlation of FIBROSpect II with histologic and morphometric evaluation of liver fibrosis in chronic hepatitis C. Clin Gastroenterol Hepatol 2008; 6:242-247.
94. Sebastiani G, Vario A, Guido M, Noventa F, Plebani M, Pistis R, Ferrari A, Alberti A. Stepwise combination algorithms of non-invasive markers to diagnose significant fibrosis in chronic hepatitis C. J Hepatol 2006; 44:686-693.
95. Imbert-Bismut F, Ratziu V, Pieroni L, Charlotte F, Benhamou Y, Poynard T. Biochemical markers of liver fibrosis in patients with hepatitis C virus infection: a prospective study. Lancet 2001; 357:1069-1075.
96. Nunes D, Fleming C, Offner G, O'Brien M, Tumilty S, Fix O, Heeren T, Koziel M, Graham C, Craven D E, Stuver S, Horsburgh C R, Jr. HIV infection does not affect the performance of noninvasive markers of fibrosis for the diagnosis of hepatitis C virus-related liver disease. J Acquir Immune Defic Syndr 2005; 40:538-544.
97. Castera L, Vergniol J, Foucher J, Le B B, Chanteloup E, Haaser M, Darriet M, Couzigou P, de L, V. Prospective comparison of transient elastography, Fibrotest, APRI, and liver biopsy for the assessment of fibrosis in chronic hepatitis C. Gastroenterology 2005; 128:343-350.
98. Guanabens N, Pares A, Alvarez L, Martinez de Osaba M J, Monegal A, Penis P, Ballesta A M, Rodes J. Collagen-related markers of bone turnover reflect the severity of liver fibrosis in patients with primary biliary cirrhosis. J Bone Miner Res 1998; 13:731-738.
99. Moller S, Hansen M, Hillingso J, Jensen J E, Henriksen J H. Elevated carboxy terminal cross linked telopeptide of type I collagen in alcoholic cirrhosis: relation to liver and kidney function and bone metabolism. Gut 1999; 44:417-423.
100. Rosen H N, Parker R A, Greenspan S L, Iloputaife I D, Bookman L, Chapin D, Perlmutter I, Kessel B, Qvist P, Rosenblatt M. Evaluation of ability of biochemical markers of bone turnover to predict a response to increased doses of HRT. Calcif Tissue Int 2004; 74:415-423.
101. Lein M, Wirth M, Miller K, Eickenberg H U, Weissbach L, Schmidt K, Haus U, Stephan C, Meissner S, Loening S A, Jung K. Serial Markers of Bone Turnover in Men with Metastatic Prostate Cancer Treated with Zoledronic Acid for Detection of Bone Metastases Progression. Eur Urol 2007.
102. Attallah A M, Toson E A, Shiha G E, Omran M M, bdel-Aziz M M, El-Dosoky I. Evaluation of serum procollagen aminoterminal propeptide III, laminin, and hydroxyproline as predictors of severe fibrosis in patients with chronic hepatitis C. J Immunoassay Immunochem 2007; 28:199-211.
103. Ulrich D, Noah E M, von H D, Pallua N. TIMP-1, MMP-2, MMP-9, and PIIINP as serum markers for skin fibrosis in patients following severe burn trauma. Plast Reconstr Surg 2003; 111:1423-1431.
104. Farkkila M, Rautiainen H, Karkkainen P, Karvonen A L, Nurmi H, Niemela O. Serological markers for monitoring disease progression in noncirrhotic primary biliary cirrhosis on ursodeoxycholic acid therapy. Liver Int 2008; 28:787-797.
105. Guechot J, Poupon R E, Giral P, Balkau B, Giboudeau J, Poupon R. Relationship between procollagen III aminoterminal propeptide and hyaluronan serum levels and histological fibrosis in primary biliary cirrhosis and chronic viral hepatitis C. J Hepatol 1994; 20:388-393.
106. Klappacher G, Franzen P, Haab D, Mehrabi M, Binder M, Plesch K, Pacher R, Grimm M, Pribill I, Eichler H G,. Measuring extracellular matrix turnover in the serum of patients with idiopathic or ischemic dilated cardiomyopathy and impact on diagnosis and prognosis. Am J Cardiol 1995; 75:913-918.
107. Imbert-Bismut F, Ratziu V, Pieroni L, Charlotte F, Benhamou Y, Poynard T. Biochemical markers of liver fibrosis in patients with hepatitis C virus infection: a prospective study. Lancet 2001; 357:1069-1075.
108. Suzuki, K., Enghild, J. J., Morodomi, T., Salvesen, G., and Nagase, H. 1990. Mechanisms of activation of tissue procollagenase by matrix metalloproteinase 3 (stromelysin). Biochemistry 29:10261-10270.
109. Lijnen, H. R. 2001. Plasmin and matrix metalloproteinases in vascular remodeling. Thromb. Haemost. 86:324-333.
Those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents the specific embodiments of the invention described herein. Such equivalents are intended to be encompassed by this invention and the following claims.

## Claims

1. A method of detecting peptide fragments comprising an N- or C-terminal neo-epitope formed by cleavage in vivo of a protein by a proteinase in a patient, said method comprising:
a. obtaining a biofluid sample from a human patient; and
b. detecting whether said peptide fragments comprising said N- or C-terminal neo-epitope are present in the biofluid sample by contacting the biofluid sample with an antibody and detecting binding between the peptide fragments and the antibody;
wherein said antibody is raised against a synthetic peptide corresponding to a C-terminal or N-terminal neo-epitope amino acid sequence formed by cleavage of a protein by a proteinase and has specific binding affinity for said C-terminal or N-terminal neo-epitope amino acid sequence, and
wherein said antibody specifically binds a neo-epitope consisting of an N-terminal amino acid sequence present in the peptide fragments produced by cleavage of collagen type I, said peptide fragments comprising an N-terminal sequence selected from the group consisting of:











Collagen I, alpha1

















ISVPGP... SEQ ID NO: 23
VPGPMG... SEQ ID NO: 24
PGPMGP... SEQ ID NO: 25


 


FQGPPG... SEQ ID NO: 27
KNGDDG... SEQ ID NO: 28
ARGLPG... SEQ ID NO: 29


 


LDGAKG... SEQ ID NO: 31
LPGERG... SEQ ID NO: 32
ATGAAG... SEQ ID NO: 67


 


VRGEPG... SEQ ID NO: 33
PGAKGA... SEQ ID NO: 34
GIAGAP... SEQ ID NO: 69


 


GGPPGP... SEQ ID NO: 35
NSGEPG... SEQ ID NO: 36
VQGPPG... SEQ ID NO: 71


 


DGVAGP... SEQ ID NO: 37
ERGSPG... SEQ ID NO: 38
RGSPGP... SEQ ID NO: 74


 


LTGSPG... SEQ ID NO: 39
QDGRPG... SEQ ID NO: 40
ARGQAG... SEQ ID NO: 77


 


RGVPGP... SEQ ID NO: 41
VGPAGK... SEQ ID NO: 42
KDGEAG... SEQ ID NO: 80


 


ERGEQG... SEQ ID NO: 43
RGEQGP... SEQ ID NO: 44
QGLPGP... SEQ ID NO: 83


 


PGERGV... SEQ ID NO: 45
ANGAPG... SEQ ID NO: 46
AGLPGP... SEQ ID NO: 86


 


ARGAPG... SEQ ID NO: 47
PGDRGE... SEQ ID NO: 48
FAGPPG... SEQ ID NO: 75


 


AKGDAG... SEQ ID NO: 49
PIGNVG... SEQ ID NO: 50
NVGAPG... SEQ ID NO: 78


 


AAGRVG... SEQ ID NO: 51
PPGPAG... SEQ ID NO: 52
GEVGPP... SEQ ID NO: 81


 


GADGPA... SEQ ID NO: 53
GPQGIA... SEQ ID NO: 54
IAGQRG... SEQ ID NO: 84


 


GQRGVV... SEQ ID NO: 55
QRGVVG... SEQ ID NO: 56
RGVVGL... SEQ ID NO: 87


 


GLPGQR... SEQ ID NO: 57
PGLPGP... SEQ ID NO: 58
EPGKQG... SEQ ID NO: 90


 


PMGPPG... SEQ ID NO: 59
MGPPGL... SEQ ID NO: 60
LAGPPG... SEQ ID NO: 89


 


DKGETG... SEQ ID NO: 61
LQGPPG... SEQ ID NO: 62
PSGASG... SEQ ID NO: 92


 


SAGAPG... SEQ ID NO: 63
RTGDAG... SEQ ID NO: 64
VGPPGP... SEQ ID NO: 99


 


FDFSF... SEQ ID NO: 65
DFSF... SEQ ID NO: 66
AGQRGV... SEQ ID NO: 95


 


LPGPGG... SEQ ID NO: 26
QAGVMG... SEQ ID NO: 76
VVGLPG... SEQ ID NO: 98


 


SGLDGA... SEQ ID NO: 30
PAGERG... SEQ ID NO: 79
GKQGPS... SEQ ID NO: 93


 


AKGEAG... SEQ ID NO: 68
ARGERG... SEQ ID NO: 82
ARGPAG... SEQ ID NO: 96


 


IAGAPG... SEQ ID NO: 70
LTGPIG... SEQ ID NO: 85
ASGPAG... SEQ ID NO: 97


 


LPGPPG... SEQ ID NO: 72
AGPPGA... SEQ ID NO: 88
GPPGPP... SEQ ID NO: 100


 


AGPKGS... SEQ ID NO: 73
ATGFPG... SEQ ID NO: 91



 



GPPGPA... SEQ ID NO: 94.








or wherein said antibody specifically binds a neo-epitope consisting of a C-terminal amino acid sequence present in the peptide fragments produced by cleavage of collagen type I, said peptide fragments comprising an C-terminal sequence selected from the group consisting of:











Collagen I, alpha1

















...QLSYGY SEQ ID NO: 101
...EKSTGG SEQ ID NO: 102
...PPGPQG SEQ ID NO: 103


 


...KGHRGF SEQ ID NO: 104
...PSGPRG SEQ ID NO: 105
...APGPQG SEQ ID NO: 106


 


...APGPAG SEQ ID NO: 107
...FPGAVG SEQ ID NO: 108
...SEGPQG SEQ ID NO: 109


 


...GANGAP SEQ ID NO: 110
...ANGAPG SEQ ID NO: 46
...SGPQGP SEQ ID NO: 112


 


...EPGPVG SEQ ID NO: 113
...EPGPTG SEQ ID NO: 114
...RGFPGA SEQ ID NO: 115


 


...KGPAGE SEQ ID NO: 116
...RGSPGP SEQ ID NO: 74
...LPGAKG SEQ ID NO: 118


 


...AVGPAG SEQ ID NO: 122
...PPGARG SEQ ID NO: 120
...PGKAGE SEQ ID NO: 121


 


...APGPDG SEQ ID NO: 125
...PAGPAG SEQ ID NO: 123
...AGPAGE SEQ ID NO: 124


 


...KDGVRG SEQ ID NO: 128
...RGERGF SEQ ID NO: 126
...PAGPRG SEQ ID NO: 127


 


...PGPAGF SEQ ID NO: 131
...PAGPTG SEQ ID NO: 129
...TGARGA SEQ ID NO: 130


 


...SAGPPG SEQ ID NO: 134
...EPGDAG SEQ ID NO: 132
...PAGPPG SEQ ID NO: 133


 


...GEVGPP SEQ ID NO: 81
...ATGFPG SEQ ID NO: 91
...NAGPPG SEQ ID NO: 136


 


...PGPQGI SEQ ID NO: 140
...GEKGSP SEQ ID NO: 138
...GAPGTP SEQ ID NO: 139


 


...IAGQRG SEQ ID NO: 84
...GPQGIA SEQ ID NO: 54
...PQGIAG SEQ ID NO: 142


 


...GPSGEP SEQ ID NO: 146
...GQRGVV SEQ ID NO55
...RGERGF SEQ ID NO: 126


 


...PVGPVG SEQ ID NO: 149
...ERGPPG SEQ ID NO147
...RGPPGP SEQ ID NO: 148


 


...EQGPSG SEQ ID NO: 152
...PQGPRG SEQ ID 150
...HRGFSG SEQ ID NO: 151


 


...GPPGPP SEQ ID NO: 100
...PRGPPG SEQ ID NO153
...PPGPRG SEQ ID NO: 154


 


...PPGPPG SEQ ID NO: 119
...GPPSAG SEQ ID NO156
...PPSAGF SEQ ID NO: 157


 


...QMGPRG SEQ ID NO: 161
...PPGPAG SEQ ID NO52
...TPGPQG SEQ ID NO: 160


 


...PGADGQ SEQ ID NO: 164
...PGPPGA SEQ ID 162
...QGIAGQ SEQ ID NO: 63


 


...PPGPKG SEQ ID NO: 167
...AGSPGF SEQ ID NO165
...LPGPSG SEQ ID NO: 166


 


...PKGPAG SEQ ID NO: 170
...PGERGA SEQ ID NO168
...PMGPPG SEQ ID NO: 59


 


...GPAGRP SEQ ID NO: 173
...PPGPIG SEQ ID NO174
...SPGEQG SEQ ID NO: 172


 


...TGDAGP SEQ ID NO: 175.








or wherein said antibody specifically binds a neo-epitope consisting of an N-terminal amino acid sequence present in the peptide fragments produced by cleavage of collagen type IV, said peptide fragments comprising an N-terminal sequence selected from the group consisting of:











Collagen type IV

















IDGYRG... SEQ ID NO: 609
MGPPGT... SEQ ID NO: 610
DGLPGS... SEQ ID NO: 611


 


PGSKGE... SEQ ID NO: 612
RGFPGP... SEQ ID NO: 613
PGPPGL... SEQ ID NO: 614


 


GPPGPP... SEQ ID NO: 100
GLPGSM... SEQ ID NO: 615
GLPGQQ... SEQ ID NO: 616


 


PGIGVQ... SEQ ID NO: 618
GIGPPG... SEQ ID NO: 611
PGLPGI... SEQ ID NO: 504


 


PGPKGF... SEQ ID NO: 621
SPGIPG... SEQ ID NO: 619
PGMQGE... SEQ ID NO: 620


 


LGSKGE... SEQ ID NO: 617
KGQPGL... SEQ ID NO: 622
RGPPGP... SEQ ID NO: 148


 


IRGEPG... SEQ ID NO: 626
FPGPPG... SEQ ID NO: 627
GPLGEK... SEQ ID NO: 625


 


DGVIGM... SEQ ID NO: 629
PGNPGI... SEQ ID NO: 630
PGLKGD... SEQ ID NO: 624


 


IKGDKG... SEQ ID NO: 631
PGSPGC... SEQ ID NO: 632
PPSDEI... SEQ ID NO: 623


 


GPPGVP... SEQ ID NO: 633
DQGDQG... SEQ ID NO: 634
GAPGPQ... SEQ ID NO: 445


 


KGSIGI... SEQ ID NO: 635
GIPGAP... SEQ ID NO: 372
DRGPQG... SEQ ID NO: 442


 


ERGSPG... SEQ ID NO: 38
LQGIRG... SEQ ID NO: 628
DGGVPN... SEQ ID NO: 638


 


SGRDGL... SEQ ID NO: 639
PGPMGP... SEQ ID NO: 25
GPPGLM... SEQ ID NO: 643


 


DGYRGP... SEQ ID NO: 642
AEGLPG... SEQ ID NO: 641
LPGFAG... SEQ ID NO: 644


 


GIPGMP... SEQ ID NO: 645
PPGRLG... SEQ ID NO: 636
GPPGEK... SEQ ID NO: 640


 


EKGQKG... SEQ ID NO: 637
.LPGPDG SEQ ID NO: 2214
.PGIPGT SEQ ID NO: 2227


 


.ILGHVP SEQ ID NO: 2212
.LRGIPG SEQ ID NO: 760
.PGDIVF SEQ ID NO: 2215


 


.PGLPGQ SEQ ID NO: 2213
.PGFPGA SEQ ID NO: 2218
.GNKGDP SEQ ID NO: 2216


 


.SGYPGN SEQ ID NO: 2217
.QQGNRG SEQ ID NO: 2221
.PGPRGK SEQ ID NO: 2219


 


.VSGPPG SEQ ID NO: 2220
.VGQPGP SEQ ID NO: 2222
.PGPPGP SEQ ID NO: 458


 


.KRGPPG SEQ ID NO: 2223
.GEPGMQ SEQ ID NO: 2224
.SKGEKG SEQ ID NO: 2226


 


.LHGFPG SEQ ID NO: 2225

.GEPGPP SEQ ID NO: 675.








wherein said antibody specifically binds a neo-epitope consisting of a C-terminal amino acid sequence present in the peptide fragments produced by cleavage of collagen type IV, said peptide fragments comprising an C-terminal sequence selected from the group consisting of:











Collagen type IV

















...RGPPGP SEQ ID NO: 148
...SVDHGF SEQ ID NO: 646
...PSVDHG SEQ ID NO: 647


 


...VDHGFL SEQ ID NO: 648
...PGQPGY SEQ ID NO: 649
...QPGYTN SEQ ID NO: 650


 


...PGLPGS SEQ ID NO: 651
...GTPSVD SEQ ID NO: 652
...SVGSPG SEQ ID NO: 653


 


...LPGSMG SEQ ID NO: 654
...GPPGVP SEQ ID NO: 633
...PGFPGL SEQ ID NO: 655


 


...PGLPGE SEQ ID NO: 656
...GDPGPP SEQ ID NO: 373
...HQGEMG SEQ ID NO: 657


 


...GPPGLV SEQ ID NO: 658
...PGIPGP SEQ ID NO: 659
...PGFPGT SEQ ID NO: 660


 


...PGLPGP SEQ ID NO: 58
...DGIPGP SEQ ID NO: 661
...SGPKGY SEQ ID NO: 662


 


...GIGPPG SEQ ID NO: 611
...PGPRGE SEQ ID NO: 663
...GQGPPG SEQ ID NO: 664


 


...PGAIGP SEQ ID NO: 668
...KVDMGS SEQ ID NO: 665
...PGIDGV SEQ ID NO: 666


 


...GPKGPP SEQ ID NO: 671
...PPGPPG SEQ ID NO: 119
...KGHMGE SEQ ID NO: 667


 


...GPKGLP SEQ ID NO: 670
...SPGPPG SEQ ID NO: 672
...KGLPGP SEQ ID NO: 669


 


...GSVGYP SEQ ID NO: 673
...PQGPPG SEQ ID NO: 389
...PPGSPG SEQ ID NO: 674


 


...GEPGPP SEQ ID NO: 675
...AGNPGP SEQ ID NO: 676
...FPGPQG SEQ ID NO: 679


 


...PGYTNG SEQ ID NO: 678
...SVDHGF SEQ ID NO: 646
...LSGPPG SEQ ID NO: 680


 


...PGPQGP SEQ ID NO: 453
...GQGPPG SEQ ID NO: 664
...PGAPGL SEQ ID NO: 467


 


...GVMGTP SEQ ID NO: 681
...PGIKGS SEQ ID NO: 677
...PGPPGP SEQ ID NO: 458


 


...GVSGPK SEQ ID NO: 682




 


HVPGML. SEQ ID NO: 2228
LPVPGQ. SEQ ID NO: 2229
LGPPGL. SEQ ID NO: 2230


 


GVPGQA. SEQ ID NO: 2231
VPGQAQ. SEQ ID NO: 2232
GPDGFL. SEQ ID NO: 2233


 


QEGPLG. SEQ ID NO: 2234
LPGEVL. SEQ ID NO: 2235
RGIPGF. SEQ ID NO: 2236


 


NRGLGF. SEQ ID NO: 2238
IPSDTL. SEQ ID NO: 2239
PAGEKG. SEQ ID NO: 2240


 


GEKGNK. SEQ ID NO: 2241
PVGPPG. SEQ ID NO: 2242
DIVFRK. SEQ ID NO: 2243


 


PPGPKG. SEQ ID NO: 167
RGKPGM. SEQ ID NO: 2244
PGTRGL. SEQ ID NO: 2245


 


GEKGSK. SEQ ID NO: 2246
EKGSKG. SEQ ID NO: 2247
SGQPGL. SEQ ID NO: 2248


 


AGIPQK. SEQ ID NO: 2237.








or wherein said antibody specifically binds a neo-epitope consisting of an N-terminal amino acid sequence present in the peptide fragments produced by cleavage of biglycan, decorin, lumican, versican, or perlecan said peptide fragments comprising an N-terminal sequence selected from the group consisting of:













Biglycan









SVPKEI... SEQ ID NO: 949
GLKLNY... SEQ ID NO: 950
RISEAK... SEQ ID NO: 951


 


NSGFEP... SEQ ID NO: 952
LKSVPK... SEQ ID NO: 953
AIELED... SEQ ID NO: 954


 


LRISEA... SEQ ID NO: 958
QCSDLG... SEQ ID NO: 955
EAKLTG... SEQ ID NO: 956


 


LLDLQN... SEQ ID NO: 961
LTGIPK... SEQ ID NO: 959
LKAVPK... SEQ ID NO: 960


 


IELEDL... SEQ ID NO: 957




 







Decorin









IVIELG... SEQ ID NO: 962
DEASGI... SEQ ID NO: 963
VNNKIS... SEQ ID NO: 964


 


NGLNQM... SEQ ID NO: 965
LHLDGN... SEQ ID NO: 966
LILVNN... SEQ ID NO: 967


 


KITEIK... SEQ ID NO: 969
GLPPSL... SEQ ID NO: 970
SNPVQY... SEQ ID NO: 971


 


SSGIEN... SEQ ID NO: 968




 







Versican









LLASDA... SEQ ID NO: 972
LATVGE... SEQ ID NO: 973
ETTVLV... SEQ ID NO: 974


 


ENQDAR... SEQ ID NO: 975
NGFDQC... SEQ ID NO: 976
SLTVVK... SEQ ID NO: 977


 







Lumican









SLEDLQ... SEQ ID NO: 978
LKEDAV... SEQ ID NO: 979
HLQHNR... SEQ ID NO: 980


 


LQHNRL... SEQ ID NO: 985




 







Perlecan









SIEYSP... SEQ ID NO: 981
LVNFTR... SEQ ID NO: 982
VSEAVV... SEQ ID NO: 983


 


EVSEAV... SEQ ID NO: 984.








or wherein said antibody specifically binds a neo-epitope consisting of a C-terminal amino acid sequence present in the peptide fragments produced by cleavage of biglycan, decorin, lumican, versican, or perlecan, said peptide fragments comprising an C-terminal sequence selected from the group consisting of:













Biglycan









...NNDISE SEQ ID NO: 986
...YWEVQP SEQ ID NO: 987
...EDLLRY SEQ ID NO: 988


 


...RISEAK SEQ ID NO: 951
...KIQAIE SEQ ID NO: 989
...PETLNE SEQ ID NO: 990


 


...LRKDDF SEQ ID NO: 991
...LLRYSK SEQ ID NO: 992
...ELRKDD SEQ ID NO: 993


 


...KDLPET SEQ ID NO: 994
...DLLRYS SEQ ID NO: 995
...AFDGLK SEQ ID NO: 996


 


...LNELHL SEQ ID NO: 997




 







Decorin









...GTNPLK SEQ ID NO: 998
...EVPDDR SEQ ID NO: 999
...GAFTPL SEQ ID NO: 1000


 


...SSGIEN SEQ ID NO: 968
...RVDAAS SEQ ID NO: 1001
...LVKLER SEQ ID 1002


 


...GMKKLS SEQ ID NO: 1003
...KDGDFK SEQ ID NO: 1004
...HLDGNK SEQ ID NO: 1005


 


...QPSTFR SEQ ID NO: 1006
...AFQGMK SEQ ID NO: 1007



 







Versican









...CDVMYG SEQ ID NO: 1008
...NGFDQC SEQ ID NO: 976
...QNGINK SEQ ID NO: 1009


 


...IGQDYK SEQ ID NO: 1010




 







Lumican









...QLTHNK SEQ ID NO: 1011
...VSAAFK 1012
...GLKSLE 1013


 







Perlecan









...EDAGSR SEQ ID NO: 1014
...EFREVS SEQ ID NO: 1015
...VAQQDS SEQ ID NO: 1016


 


...SAKEFR SEQ ID NO: 1017
...LEPEYR SEQ ID NO: 1018.








or wherein said antibody specifically binds a neo-epitope consisting of an N-terminal amino acid sequence present in the peptide fragments produced by cleavage of C-reactive protein, said peptide fragments comprising an N-terminal sequence selected from the group consisting of:











CRP

















AFVFPK SEQ ID NO: 1033
SFGGNF SEQ ID NO: 1305
FGQTDM SEQ ID NO: 1306


 


VSLKAP SEQ ID NO: 1172
KAFVFP SEQ ID NO: 1167
EVFTKP SEQ ID NO: 1201


 


TDMSRK SEQ ID NO: 1307
MSRKAF SEQ ID NO: 1308
SRKAFV SEQ ID NO: 1309


 


VFPKES SEQ ID NO: 1310
FPKESD SEQ ID NO: 1311
KESDTS SEQ ID NO: 1312


 


LSSTRG SEQ ID NO: 1313
SSTRGY SEQ ID NO: 1314
STRGYS SEQ ID NO: 1080


 


KRQDNE SEQ ID NO: 1315
WSKDIG SEQ ID NO: 1316
SKDIGY SEQ ID NO: 1317


 


SIILGQ SEQ ID NO: 1318
YLGGPF SEQ ID NO: 1322
ILGQEQ SEQ ID NO: 1320


 


IYLGGP SEQ ID NO: 1321
KYEVQG SEQ ID NO: 1294
LGGPFS SEQ ID NO: 1323


 


ALKYEV SEQ ID NO: 1133
YTELSS SEQ ID NO: 1325
VQGEVF SEQ ID NO: 1324


 


KPQLWP SEQ ID NO: 1157
QEQDSF SEQ ID NO: 1328
ILIFWS SEQ ID NO: 1326


 


LVGDIG SEQ ID NO: 1327
QDSFGG SEQ ID NO: 1331
RGYSIF SEQ ID NO: 1329


 


GAEASI SEQ ID NO: 1330
DTSYVS SEQ ID NO: 1334
TIYLGG SEQ ID NO: 1332


 


EINTIY SEQ ID NO: 1333
SPDEIN SEQ ID NO: 1337
AEASII SEQ ID NO: 1335


 


TSWESA SEQ ID NO: 1336
FVLSPD SEQ ID NO: 1340
GGPFSP SEQ ID NO: 1338


 


YEVQGE SEQ ID NO: 1339
IVEFWV SEQ ID NO: 1343
LKKGYT SEQ ID NO: 1341


 


IILGQE SEQ ID NO: 1319
EVQGEV SEQ ID NO: 1346
ESDTSY SEQ ID NO: 1344


 


RKAFVF SEQ ID NO: 1342
SLKAPL SEQ ID NO: 1222
FVFPK SEQ ID NO: 1059


 


SDTSYV SEQ ID NO: 1347
SYATKR SEQ ID NO: 1349
LKAPLT SEQ ID NO: 1173


 


IFSYAT SEQ ID NO: 1348
WVDGKP SEQ ID NO: 1352
YATKRQ SEQ ID NO: 1350


 


EFWVDG SEQ ID NO: 1351
GQEQDS SEQ ID NO: 1355
VDGKPR SEQ ID NO: 1353


 


LGQEQD SEQ ID NO: 1354
LNWRA SEQ ID NO: 1127
QSLVGD SEQ ID NO: 1356


 


SPNVLN SEQ ID NO: 1357
GEVFTK SEQ ID NO: 1359
LNWRAL SEQ ID NO: 1128


 


QGEVFT SEQ ID NO: 1358
VRKSLK SEQ ID NO: 1205
VFTKPQ SEQ ID NO: 1153


 


IFWSKD SEQ ID NO: 1181
ATKRQD SEQ ID NO: 1362
KKGYTV SEQ ID NO: 1360


 


FSYATK SEQ ID NO: 1361
NWRAL SEQ ID NO: 1253
FWSKDI SEQ ID NO: 1363


 


VLNWRA SEQ ID NO: 1249
DFVLSP SEQ ID NO: 1366
NWRALK SEQ ID NO: 1364


 


GSQSLV SEQ ID NO: 1365
INTIYL SEQ ID NO: 1372
VLSPDE SEQ ID NO: 1367


 


LKYEVQ SEQ ID NO: 1134
WRALKY SEQ ID NO: 1375
KSLKKG SEQ ID NO: 1371


 


GNFEGS SEQ ID NO: 1369
TKPQLW SEQ ID NO: 1345
RALKYE SEQ ID NO: 1373


 


FVFPKE SEQ ID NO: 1060
KGYTVG SEQ ID NO: 1296
GPFSPN SEQ ID NO: 1376


 


FYTELS SEQ ID NO: 1374
TKRQDN SEQ ID NO: 1368
SLKKGY SEQ ID NO: 1370.








or wherein said antibody specifically binds a neo-epitope consisting of a C-terminal amino acid sequence present in the peptide fragments produced by cleavage of C-reactive protein, said peptide fragments comprising an C-terminal sequence selected from the group consisting of:











CRP

















AFVFPK SEQ ID NO: 1033
KPQLWP SEQ ID NO: 1157
PDEINT SEQ ID NO: 1377


 


KESDTS SEQ ID NO: 1312
DSFGGN SEQ ID NO: 1378
VFTKPQ SEQ ID NO: 1153


 


LTKPLK SEQ ID NO: 1379
SDTSYV SEQ ID NO: 1347
DTSYVS SEQ ID NO: 1334


 


KDIGYS SEQ ID NO: 1380
STRGYS SEQ ID NO: 1080
YATKRQ SEQ ID NO: 1350


 


GAEASI SEQ ID NO: 1330
DGKPRV SEQ ID NO: 1381
VDGKPR SEQ ID NO: 1353


 


SPNVLN SEQ ID NO: 1357
GPFSPN SEQ ID NO: 1376
NFEGSQ SEQ ID NO: 1382


 


ALKYEV SEQ ID NO: 1133
YEVQGE SEQ ID NO: 1339
LKYEVQ SEQ ID NO: 1134


 


KAFVFP SEQ ID NO: 1167
VSLKAP SEQ ID NO: 1172
LKAPLT SEQ ID NO: 1173


 


LKKGYT SEQ ID NO: 1341
LGQEQD SEQ ID NO: 1354
NVNMWD SEQ ID NO: 1383


 


PRVRKS SEQ ID NO: 1384
TVGSEI SEQ ID NO: 1385
SRKAFV SEQ ID NO: 1309


 


GYSFTV SEQ ID NO: 1386
IILGQE SEQ ID NO: 1319
EGSQSL SEQ ID NO: 1387


 


INTIYL SEQ ID NO: 1372
WSKDIG SEQ ID NO: 1316
EQDSFG SEQ ID NO: 1388


 


NVLNWR SEQ ID NO: 1389
ASGIVE SEQ ID NO: 1390
NTIYLG SEQ ID NO: 1391


 


VGAEAS SEQ ID NO: 1392
TDMSRK SEQ ID NO: 1307
FVFPKE SEQ ID NO: 1060


 


QTDMSR SEQ ID NO: 1394
TSYVSL SEQ ID NO: 1396
MSRKAF SEQ ID NO: 1308


 


PKESDT SEQ ID NO: 1395
QDNEIL SEQ ID NO: 1398
ESDTSY SEQ ID NO: 1344


 


DNEILI SEQ ID NO: 1397
TVGAEA SEQ ID NO: 1401
NEILIF SEQ ID NO: 1399


 


YTVGAE SEQ ID NO: 1400
GNFEGS SEQ ID NO: 1369
DIGNVN SEQ ID NO: 1404


 


FEGSQS SEQ ID NO: 1403
LNWRAL SEQ ID NO: 1128
NWRALK SEQ ID NO: 1364


 


LNWRA SEQ ID NO: 1127
EVFTKP SEQ ID NO: 1201
VFTKPQ SEQ ID NO: 1153


 


KYEVQG SEQ ID NO: 1294
RQDNEI SEQ ID NO: 1405
IFWSKD SEQ ID NO: 1181


 


KRQDNE SEQ ID NO: 1315
VRKSLK SEQ ID NO: 1205
SYVSLK SEQ ID NO: 1407


 


FTKPQL SEQ ID NO: 1406
TKPLKA SEQ ID NO: 1408
SIFSYA SEQ ID NO: 1409


 


SLKAPL SEQ ID NO: 1222
GNVNMW SEQ ID NO: 1410
SQSLVG SEQ ID NO: 1411


 


GSQSLV SEQ ID NO: 1365
GGNFEG SEQ ID NO: 1413
LVGDIG SEQ ID NO: 1327


 


FGGNFE SEQ ID NO: 1412
RALKYE SEQ ID NO: 1373
KKGYTV SEQ ID NO: 1360


 


VQGEVF SEQ ID NO: 1324
GKPRVR SEQ ID NO: 1414
PFSPNV SEQ ID NO: 1402


 


DMSRKA SEQ ID NO: 1415
EILIFW SEQ ID NO: 1416
QGEVFT SEQ ID NO: 1358


 


FPKESD SEQ ID NO: 1311
IGYSFT SEQ ID NO: 1417
SPDEIN SEQ ID NO: 1337


 


APLTKP SEQ ID NO: 1393
FWSKDI SEQ ID NO: 1363.








or wherein said antibody specifically binds a neo-epitope consisting of an N-terminal amino acid sequence present in the peptide fragments produced by cleavage of elastin, said peptide fragments comprising an N-terminal sequence selected from the group consisting of:











Elastin

















GVPGAI SEQ ID NO: 1898
AIPGGV SEQ ID NO: 1899
GVPGGV SEQ ID NO: 1900


 


ALGGGA SEQ ID NO: 1901
LGGGAL SEQ ID NO: 1902
GGALGP SEQ ID NO: 1903


 


PLKPVP SEQ ID NO: 1904
LKPVPG SEQ ID NO: 1905
GLAGAG SEQ ID NO: 1906


 


GLGAGL SEQ ID NO: 1907
LGAGLG SEQ ID NO: 1908
AGLGAF SEQ ID NO: 1909


 


LVPGGV SEQ ID NO: 1910
VADAAA SEQ ID NO: 1911
KAAKAG SEQ ID NO: 1912


 


PVGYPG SEQ ID NO: 1913
ARFPGV SEQ ID NO: 1914
RFPGVG SEQ ID NO: 1915


 


VGPFGG SEQ ID NO: 1916
GPQPGV SEQ ID NO: 1917
PQPGVP SEQ ID NO: 1918


 


TTGKLP SEQ ID NO: 1919
LPYGYG SEQ ID NO: 1920
GYGPGG SEQ ID NO: 1921


 


FGAGAA SEQ ID NO: 1922
GVLPGV SEQ ID NO: 1923
VLPGVG SEQ ID NO: 1924


 


TPAAAA SEQ ID NO: 1928
VPGAIP SEQ ID NO: 1926
AIPGIG SEQ ID NO: 1927


 


VGVPGA SEQ ID NO: 1931
PAAAAA SEQ ID NO: 1929
AAAAAA SEQ ID NO: 1930


 


GIPVVP SEQ ID NO: 1934
AVGPGV SEQ ID NO: 1932
GVPGVG SEQ ID NO: 1933


 


ARPGVG SEQ ID NO: 1937
IPGAAV SEQ ID NO: 1935
GAAVPG SEQ ID NO: 1936


 


SVGGVP SEQ ID NO: 1940
RPGVGV SEQ ID NO: 1938
VGGIPT SEQ ID NO: 1939


 


VGVAPG SEQ ID NO: 1943
VGGVPG SEQ ID NO: 1941
GVGTPA SEQ ID NO: 1942


 


GVPVAP SEQ ID NO: 1946
VAPGVG SEQ ID NO: 1944
GVAPGV SEQ ID NO: 1945


 


LGAGIP SEQ ID NO: 1949
GAGIPG SEQ ID NO: 1947
PGGVAA SEQ ID NO: 1948


 


PGALAA SEQ ID NO: 1952
AKYGAA SEQ ID NO: 1953
AGIPGL SEQ ID NO: 1925


 


ALGGVG SEQ ID NO: 1955
LGGVGI SEQ ID NO: 1956
YGAAVP SEQ ID NO: 1954


 


AGPAAA SEQ ID NO: 1958
GPAAAA SEQ ID NO: 1959
GVGIPG SEQ ID NO: 1957


 


GLGVPG SEQ ID NO: 1961
LGGIPP SEQ ID NO: 1962
FGLVGA SEQ ID NO: 1960


 


PLGGVA SEQ ID NO: 1964
LGGVAA SEQ ID NO: 1965
LGGVLG SEQ ID NO: 1963


 


GVGLPG SEQ ID NO: 1967
VGLPGV SEQ ID NO: 1968
GGVAAR SEQ ID NO: 1966


 


VPGVPV SEQ ID NO: 1970
VPGVGI SEQ ID NO: 1971
LPGVYP SEQ ID NO: 1969


 


APGVGV SEQ ID NO: 1973
VPGGVA SEQ ID NO: 1974
VGISPE SEQ ID NO: 1972


 


VPGGVF SEQ ID NO: 1979
YPTGTG SEQ ID NO: 1977
YPGGVL SEQ ID NO: 1975


 


LGPGGK SEQ ID NO: 1982
GVFYPG SEQ ID NO: 1980
VPGAGV SEQ ID NO: 1978


 


LAGAGL SEQ ID NO: 1985
GPGGKP SEQ ID NO: 1983
VFYPGA SEQ ID NO: 1981


 


LGAFPA SEQ ID NO: 1988
AGAGLG SEQ ID NO: 1986
PGGKPL SEQ ID NO: 1984


 


LGVSAG SEQ ID NO: 1991
AFPAVT SEQ ID NO: 1989
GAGLGA SEQ ID NO: 1987


 


FPGVGV SEQ ID NO: 1994
VPGVGL SEQ ID NO: 1992
AVTFPG SEQ ID NO: 1990


 


PGVPLG SEQ ID NO: 1996
KPGAPG SEQ ID NO: 1995
PGVGLP SEQ ID NO: 1993


 


YGPGGV SEQ ID NO: 1999
GYPIKA SEQ ID NO: 1997
PKAPGV SEQ ID NO: 1493


 


GAGVPG SEQ ID NO: 2002
AGYPTGSEQ ID NO: 2000
PKLPGG SEQ ID NO: 1998


 


IPGIGG SEQ ID NO: 2005
AGVPGV SEQ ID NO: 2003
TGVGPQ SEQ ID NO: 2001


 


GPGFGP SEQ ID NO: 1976
IGGIAG SEQ ID NO: 2006
GVPGVP SEQ ID NO: 2004


 


VPGVGV SEQ ID NO: 2008
PGFGPG SEQ ID NO: 1950
GIAGVG SEQ ID NO: 2007


 


AVPGVV SEQ ID NO: 2011
VPVGVA SEQ ID NO: 2009
PGVVGV SEQ ID NO: 1951


 


GVGAGG SEQ ID NO: 2014
VPGVVS SEQ ID NO: 2012
VPGAGI SEQ ID NO: 2010


 


FGLVPG SEQ ID NO: 2017
VGAGGF SEQ ID NO: 2015
YGARPG SEQ ID NO: 2013


 


PGVGLA SEQ ID NO: 2020
GLVPGV SEQ ID NO: 2018
VGVGGI SEQ ID NO: 2016


 


LRAAAG SEQ ID NO: 2023
GVGLAP SEQ ID NO: 2021
LVPGVG SEQ ID NO: 2019


 


GIPGLG SEQ ID NO: 2026
LVGAAG SEQ ID NO: 2024
VGLAPG SEQ ID NO: 2022


 


VGIPGG SEQ ID NO: 2032
LGVGVG SEQ ID NO: 2027
LVPGGP SEQ ID NO: 2025


 


GLVGAA SEQ ID NO: 2035
AVPGVL SEQ ID NO: 2030
VGVPGL SEQ ID NO: 2028


 


GGVLGG SEQ ID NO: 2038
IPGGVV SEQ ID NO: 2033
VLGGLG SEQ ID NO: 2031


 


GVAARP SEQ ID NO: 2041
VGAAGL SEQ ID NO: 2036
VVGAGP SEQ ID NO: 2034


 


GVYPGG SEQ ID NO: 2044
GAGQFP SEQ ID NO: 2039
LGGLGV SEQ ID NO: 2037


 


AAVPGV SEQ ID NO: 2047
VAARPG SEQ ID NO: 2042
AFQFPL SEQ ID NO: 2041


 


VPGVLG SEQ ID NO: 2050
AAGLGG SEQ ID NO: 2048
RPGFGL SEQ ID NO: 2043


 


ISPEAQ SEQ ID NO: 2053
GQFPLG SEQ ID NO: 2051
FGPGVV SEQ ID NO: 2046


 


LGALGG SEQ ID NO: 2056
GVLPGA SEQ ID NO: 2054
FPGALV SEQ ID NO: 2049


 


GKPLKP SEQ ID NO: 2059
PQAAAA SEQ ID NO: 2077
ALVPGG SEQ ID NO: 2052


 


AGLGAG SEQ ID NO: 2062
VPGVPG SEQ ID NO: 2057
AAAGLG SEQ ID NO: 2029


 


VTFPGA SEQ ID NO: 2065
IAGVGT SEQ ID NO: 2060
VGAGVP SEQ ID NO: 2055


 


GLPGVY SEQ ID NO: 2068
VVGVPG SEQ ID NO: 2063
GGLGAL SEQ ID NO: 2058


 


GAFAGI SEQ ID NO: 2071
AGIPVV SEQ ID NO: 2066
VGAGPA SEQ ID NO: 2061


 


YTTGKL SEQ ID NO: 2074
GARPGV SEQ ID NO: 2069
LGVGGL SEQ ID NO: 2064


 


PGVGVA SEQ ID NO: 2075
VAGVPS SEQ ID NO: 2072
FPLGGV SEQ ID NO: 2067


 


LAPGVG SEQ ID NO: 2078
VGTPAA SEQ ID NO: 2076
PGFGLS SEQ ID NO: 2070


 


LPYTTG SEQ ID NO: 2045
GPGVVG SEQ ID NO: 2073.








or wherein said antibody specifically binds a neo-epitope consisting of a C-terminal amino acid sequence present in the peptide fragments produced by cleavage of elastin, said peptide fragments comprising an C-terminal sequence selected from the group consisting of:











Elastin

















PGGVPG SEQ ID NO: 2079
PGAGLG SEQ ID NO: 2080
GAGLGA SEQ ID NO: 1987


 


GALGGG SEQ ID NO: 2081
LGGGAL SEQ ID NO: 1902
GGGALG SEQ ID NO: 2082


 


GLGAFP SEQ ID NO: 2083
LGAFPA SEQ ID NO: 1988
LGAGLG SEQ ID NO: 1908


 


VPGGVA SEQ ID NO: 1974
ADAAAA SEQ ID NO: 2084
VPTGAG SEQ ID NO: 2087


 


RFPGVG SEQ ID NO: 1915
VGVLPG SEQ ID NO: 2086
VPLGYP SEQ ID NO: 2089


 


PKAPGV SEQ ID NO: 1493
GVGPFG SEQ ID NO: 2088
IPGIGG SEQ ID NO: 2005


 


LPYGYG SEQ ID NO: 1920
AAAAAK SEQ ID NO: 2090
IGGIAG SEQ ID NO: 2006


 


GIAGVG SEQ ID NO: 2007
AIPGIG SEQ ID NO: 1927
GVPGVG SEQ ID NO: 1933


 


AAAAKA SEQ ID NO: 2091
VVGVPG SEQ ID NO: 2063
VVSPEA SEQ ID NO: 2094


 


GVGVPG SEQ ID NO: 2092
PVVPGA SEQ ID NO: 2093
GIPTYG SEQ ID NO: 2096


 


VGGIPT SEQ ID NO: 1939
GGIPTY SEQ ID NO: 2095
GVGAGG SEQ ID NO: 2014


 


GVGVGG SEQ ID NO: 2097
GVGGIP SEQ ID NO: 2098
VAPGVG SEQ ID NO: 1944


 


GFPGFG SEQ ID NO: 2099
FGVGVG SEQ ID NO: 2100
APGVGL SEQ ID NO: 2104


 


PGVGIS SEQ ID NO: 2101
SPEAQA SEQ ID NO: 2102
VAPGIG SEQ ID NO: 2107


 


PGVGVA SEQ ID NO: 2075
GVGVAP SEQ ID NO: 2103
AAGLGA SEQ ID NO: 2109


 


GIGPGG SEQ ID NO: 2105
APGIGP SEQ ID NO: 2106
AGVPGL SEQ ID NO: 2112


 


RAAAGL SEQ ID NO: 2108
AAAGLG SEQ ID NO: 2029
GAGPAA SEQ ID NO: 2113


 


GVPGLG SEQ ID NO: 2110
GVPGFG SEQ ID NO: 2111
GLGGLG SEQ ID NO: 2115


 


VLGGLG SEQ ID NO: 2031
VGIPGG SEQ ID NO: 2032
PAAAAK SEQ ID NO: 2117


 


PAAAAA SEQ ID NO: 1929
VPGVGG SEQ ID NO: 2114
AARPGF SEQ ID NO: 2118


 


GLGVPG SEQ ID NO: 1961
PGVGGL SEQ ID NO: 2116
GPGIPG SEQ ID NO: 2121


 


RPGFGL SEQ ID NO: 2043
GVAARP SEQ ID NO: 2041
ALGPGG SEQ ID NO: 2124


 


LSPIFP SEQ ID NO: 2119
AQAAAA SEQ ID NO: 2120
KPVPGG SEQ ID NO: 2127


 


GPGGVA SEQ ID NO: 2122
LGALGG SEQ ID NO: 2056
VTFPGA SEQ ID NO: 2065


 


GLGALG SEQ ID NO: 2123
PGAIPG SEQ ID NO: 2146
VPQPGA SEQ ID NO: 2130


 


GALGPG SEQ ID NO: 2125
TPAAAA SEQ ID NO: 1928
AGVKPK SEQ ID NO: 2133


 


AFPAVT SEQ ID NO: 1989
AVTFPG SEQ ID NO: 1990
YGYGPG SEQ ID NO: 2136


 


AAAAYK SEQ ID NO: 2128
AAKAGA SEQ ID NO: 2129
GVGTPA SEQ ID NO: 1942


 


PGVPTG SEQ ID NO: 2131
GVPTGA SEQ ID NO: 2132
AAAAAA SEQ ID NO: 1930


 


PIKAPK SEQ ID NO: 2134
KLPGGY SEQ ID NO: 2135
GVPGAG SEQ ID NO: 2140


 


VGTPAA SEQ ID NO: 2076
VPGVPG SEQ ID NO: 2057
ARPGVG SEQ ID NO: 1937


 


GIGGIA SEQ ID NO: 2137
GTPAAA SEQ ID NO: 2138
YGVGAG SEQ ID NO: 2144


 


IPVVPG SEQ ID NO: 2139
VGVPGA SEQ ID NO: 1931
VGAGGF SEQ ID NO: 2015


 


GAGIPG SEQ ID NO: 1947
PEAAAK SEQ ID NO: 2141
GISPEA SEQ ID NO: 2149


 


IPTYGV SEQ ID NO: 2145
TYGVGA SEQ ID NO: 2143
VPGAPG SEQ ID NO: 2150


 


AGGFPG SEQ ID NO: 2147
PTYGVG SEQ ID NO: 2142
APGVGV SEQ ID NO: 1973


 


VGVAPG SEQ ID NO: 1943
GIPGVA SEQ ID NO: 2148
PGIGPG SEQ ID NO: 2152


 


PGVGLA SEQ ID NO: 2020
VGLAPG SEQ ID NO: 2022
LGVGVG SEQ ID NO: 2027


 


GAGVPG SEQ ID NO: 2002
LAPGVG SEQ ID NO: 2078
LGGLGA SEQ ID NO: 2157


 


PGVGVG SEQ ID NO: 2169
GGVAAA SEQ ID NO: 2151
GPAAAA SEQ ID NO: 1959


 


GLGVGG SEQ ID NO: 2153
PGLGVG SEQ ID NO: 2154
GVGGLG SEQ ID NO: 2159


 


GLGVGA SEQ ID NO: 2155
ALAAAK SEQ ID NO: 2156
VAARPG SEQ ID NO: 2042


 


VGAGPA SEQ ID NO: 2061
AGPAAA SEQ ID NO: 1958
IFPGGA SEQ ID NO: 2163


 


GGLGVG SEQ ID NO: 2158
LGVGGL SEQ ID NO: 2064
AAKAAK SEQ ID NO: 2166


 


AGQFPL SEQ ID NO: 2160
GGVAAR SEQ ID NO: 1966
PGGVAA SEQ ID NO: 1948


 


GFGLSP SEQ ID NO: 2161
PIFPGG SEQ ID NO: 2162
VGVPGL SEQ ID NO: 2028


 


AAKFGA SEQ ID NO: 2164
AAKYGA SEQ ID NO: 2165
PGVLGG SEQ ID NO: 2085


 


AGLGAL SEQ ID NO: 2167
RPGVGV SEQ ID NO: 1938
GIPGGV SEQ ID NO: 2168


 


LKPVPG SEQ ID NO: 1905
GGFPGF SEQ ID NO: 2173
IPPAAA SEQ ID NO: 2170


 


ALVPGG SEQ ID NO: 2052
VGGVPG SEQ ID NO: 1941
ARPGFG SEQ ID NO: 2171


 


VLPGAR SEQ ID NO: 2172
GVAPGV SEQ ID NO: 1945
GLSPIF SEQ ID NO: 2174


 


PTGAGV SEQ ID NO: 2175
VAPVGV SEQ ID NO: 2126
GIPVVP SEQ ID NO: 1934


 


AGAAGK SEQ ID NO: 2176.








or wherein said antibody specifically binds a neo-epitope consisting of an N-terminal amino acid sequence present in the peptide fragments produced by cleavage of collagen type V, said peptide fragments comprising an N-terminal sequence selected from the group consisting of:











Collagen type V

















GDPGPP SEQ ID NO: 373
LRGIPG SEQ ID NO: 760
IGPPGI SEQ ID NO: 761


 


LQGPPG SEQ ID NO: 62
IGSLGH SEQ ID NO: 762
IRGPPG SEQ ID NO: 763


 


ANGSPG SEQ ID NO: 764
LIGTPG SEQ ID NO: 765
LPGEPG SEQ ID NO: 766


 


IPGRPG SEQ ID NO: 767
GPDGPP SEQ ID NO: 768
QPGPSG SEQ ID NO: 769


 


LKGNEG SEQ ID NO: 770
ERGHPG SEQ ID NO: 771
GPPGEQ SEQ ID NO: 772


 


FPGPKG SEQ ID NO: 491
PFRFGG SEQ ID NO: 773
ESQAQA SEQ ID NO: 774


 


LLGPKG SEQ ID NO: 775
QQGNPG SEQ ID NO: 776
KEGPPG SEQ ID NO: 777


 


IGLIGP SEQ ID NO: 778
GPPGPP SEQ ID NO: 100
LGPPGE SEQ ID NO: 779


 


GPPGPK SEQ ID NO: 780
SLGHPG SEQ ID NO: 781
KDGIPG SEQ ID NO: 782


 


PVGEPG SEQ ID NO: 783
LRGIPG SEQ ID NO: 760
KDGIPG SEQ ID NO: 782


 


ANGSPG SEQ ID NO: 764
LIGTPG SEQ ID NO: 765
PGEPGP SEQ ID NO: 784


 


KDGIP SEQ ID NO: 813
VLGPQG SEQ ID NO: 814
DGIPGP SEQ ID NO: 661


 


QMGPPG SEQ ID NO: 802
LLGAPG SEQ ID NO: 785
GLPGLE SEQ ID NO: 786


 


ALRGPA SEQ ID NO: 810
LALRGP SEQ ID NO: 788
LTGRPG SEQ ID NO: 789


 


ETGFQG SEQ ID NO: 808
LRGFPG SEQ ID NO: 790
KTGPIG SEQ ID NO: 791


 


EAGHPG SEQ ID NO: 811
PGPKGN SEQ ID NO: 527
EQGLPG SEQ ID NO: 793


 


GSKGPM SEQ ID NO: 809
AIGGPP SEQ ID NO: 794
GPNGDP SEQ ID NO: 795


 


EKGHPG SEQ ID NO: 812
LLGPRG SEQ ID NO: 797
GPDGPP SEQ ID NO: 768


 


GPKGSI SEQ ID NO: 816
GPKGDP SEQ ID NO: 799
GDPGPP SEQ ID NO: 373


 


ELGFQG SEQ ID NO: 817
LQGPPG SEQ ID NO: 62
EKGHIG SEQ ID NO: 801


 


LRGPAG SEQ ID NO: 818
SRGERG SEQ ID NO: 803
EKGKSG SEQ ID NO: 804


 


EDGERG SEQ ID NO: 815
LPGEPG SEQ ID NO: 766
PQGAIG SEQ ID NO: 806


 


GIPGEK SEQ ID NO: 787
PPGRPG SEQ ID NO: 792
PGPPGE SEQ ID NO: 796


 


LLGPKG SEQ ID NO: 775
GPPGEK SEQ ID NO: 640
ERGPNG SEQ ID NO: 798


 


GPMGLT SEQ ID NO: 807
GPIGEK SEQ ID NO: 805
PGIPGE SEQ ID NO: 800


 


QMGPPG SEQ ID NO: 802.








or wherein said antibody specifically binds a neo-epitope consisting of a C-terminal amino acid sequence present in the peptide fragments produced by cleavage of collagen type V, said peptide fragments comprising a C-terminal sequence selected from the group consisting of:











Collagen type V

















PIGSLG SEQ ID NO: 819
PVGEPG SEQ ID NO: 783
PGPPGE SEQ ID NO: 796


 


PPGPTG SEQ ID NO: 820
PKGPPG SEQ ID NO: 821
VAGPLG SEQ ID NO: 822


 


RPGVTG SEQ ID NO: 823
MMPFQF SEQ ID NO: 824
PGAAGP SEQ ID NO: 825


 


PVGPAG SEQ ID NO: 826
HEGPTG SEQ ID NO: 827
EPGPRG SEQ ID NO: 516


 


GPPGLP SEQ ID NO: 828
GQGPPG SEQ ID NO: 664
PPGPPG SEQ ID NO: 119


 


AGSPGE SEQ ID NO: 829
PVGALG SEQ ID NO: 830
EQGLPG SEQ ID NO: 793


 


YPGPRG SEQ ID NO: 831
HPGQRG SEQ ID NO: 832
GGGGDA SEQ ID NO: 833


 


LIGPPG SEQ ID NO: 834
GLPGLK SEQ ID NO: 835
PPGPPG SEQ ID NO: 119


 


PPGPIG SEQ ID NO: 174
KGLPGL SEQ ID NO: 836
PGPPGP SEQ ID NO: 458


 


QMGPPG SEQ ID NO: 802
LLGAPG SEQ ID NO: 785
RPGVTG SEQ ID NO: 823


 


QQARLA SEQ ID NO: 837
PPGHPG SEQ ID NO: 838
PQGPRG SEQ ID NO: 150


 


PLGPPG SEQ ID NO: 839
GEPGLL SEQ ID NO: 840
EVGPLG SEQ ID NO: 841


 


IMMPFQ SEQ ID NO: 842
GLPGLK SEQ ID NO: 835
PLGPSG SEQ ID NO: 843


 


AAGPSG SEQ ID NO: 844
PPGPLG SEQ ID NO: 845
PPGSRG SEQ ID NO: 846


 


PAGPMG SEQ ID NO: 847
PPGSGG SEQ ID NO: 848
PPGPPG SEQ ID NO: 119


 


GLPGPV SEQ ID NO: 849
LPGPVG SEQ ID NO: 850
KPGPDG SEQ ID NO: 851


 


LPGPQG SEQ ID NO: 852
VIQPLP SEQ ID NO: 853
RGPNGP SEQ ID NO: 854


 


PGPQGS SEQ ID NO: 855
EKGPLG SEQ ID NO: 856
PLGPPG SEQ ID NO: 839


 


PPGPLG SEQ ID NO: 845
TGPKGE SEQ ID NO: 858
GPKGSI SEQ ID NO: 816


 


GPPGAA SEQ ID NO: 857
PPGPAG SEQ ID NO: 52
PPGPTG SEQ ID NO: 820


 


LLGAPG SEQ ID NO: 785
PLGPLG SEQ ID NO: 863
PPGSRG SEQ ID NO: 846


 


PSGLPG SEQ ID NO: 860
LPGPPG SEQ ID NO: 72
KPGPRG SEQ ID NO: 862


 


PPGPAG SEQ ID NO: 52
PPGLPG SEQ ID NO: 861
PKGPPG SEQ ID NO: 821


 



QGLPGL SEQ ID NO: 859
PPGSRG SEQ ID NO: 846.








or wherein said antibody specifically binds a neo-epitope consisting of a N-terminal amino acid sequence present in the peptide fragments produced by cleavage of vimentin, said peptide fragments comprising an N-terminal sequence selected from the group consisting of:














Vimentin



















LLQDSV SEQ ID NO: 2182



 



FADLSE SEQ ID NO: 2183



 



ISLPLP SEQ ID NO: 2184








or wherein said antibody specifically binds a neo-epitope consisting of a C-terminal amino acid sequence present in the peptide fragments produced by cleavage of vimentin, said peptide fragments comprising a C-terminal sequence selected from the group consisting of:











Vimentin
















SVPGVR SEQ ID NO: 2185
SVPGVL SEQ ID NO: 2186.

2. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments of CRP comprising an N-terminal sequence KAFVFP . . . (SEQ ID NO: 1167).

3. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments fragments of Collagen I, alpha 1 comprising the C-terminal amino acid sequence . . . KDGVRG (SEQ ID NO: 128).

4. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments of Collagen type IV comprising the C-terminal amino acid sequence . . . VDHGFL (SEQ ID NO: 648).

5. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments fragments of Collagen type IV comprising the N-terminal amino acid sequence ILGHVP . . . (SEQ ID NO: 2212).

6. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments fragments of Collagen type IV comprising the N-terminal amino acid sequence PGDIVF . . . (SEQ ID NO: 2215).

7. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments fragments of Elastin comprising the C-terminal amino acid sequence . . . GISPEA (SEQ ID NO: 2149).

8. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments fragments of Elastin comprising the C-terminal amino acid sequence . . . GIGPGG (SEQ ID NO: 2105).

9. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments fragments of Vimentin comprising the C-terminal amino acid sequence . . . SVPGVR (SEQ ID NO: 2185).

10. A method as claimed in claim 1, wherein said antibody specifically binds the peptide fragments fragments of Vimentin comprising the C-terminal amino acid sequence . . . SVPGVL (SEQ ID NO: 2186).