title: A novel assay for extracellular matrix remodeling associated with liver fibrosis: An enzyme-linked immunosorbent assay (ELISA) for a MMP-9 proteolytically revealed neo-epitope of type III collagen.
abstract: OBJECTIVE
  Accumulation of extracellular matrix (ECM) components and increased matrix-metalloprotease (MMPs) activity are hallmarks of fibrosis. We developed an ELISA for quantification of MMP-9 derived collagen type III (CO3) degradation.
  
  
  METHODS
  A monoclonal antibody targeting a specific MMP-9 cleaved fragment of CO3 was used for development of a competitive ELISA. The assay was investigated in serum and tissues from bile duct ligated rats (BDL).
  
  
  RESULTS
  The ELISA showed no cross-reaction with either intact CO3, or other collagens. The intra- and inter-assay CV were below 10%. Liver fibrosis was demonstrated in BDL animals by semi quantitative scoring (P<0.0001). Serum levels of CO3-610 increased 2.5 fold in BDL animals (P<0.001). The CO3-610 levels were 5 fold higher in ex vivo cultures of fibrotic livers compared to controls (P<0.001).
  
  
  CONCLUSIONS
  We have developed a novel ELISA for measuring a specific fragment CO3 generated by MMP-9 important in pathogenesis of liver fibrosis.
authors.affiliation: Nordic Bioscience A/S, Herlev, Denmark. nb@nordicbioscience.com
authors.forename: N
authors.initials: N
authors.lastname: Barascuk
authors.forename: S S
authors.initials: SS
authors.lastname: Veidal
authors.forename: L
authors.initials: L
authors.lastname: Larsen
authors.forename: D V
authors.initials: DV
authors.lastname: Larsen
authors.forename: M R
authors.initials: MR
authors.lastname: Larsen
authors.forename: J
authors.initials: J
authors.lastname: Wang
authors.forename: Q
authors.initials: Q
authors.lastname: Zheng
authors.forename: R
authors.initials: R
authors.lastname: Xing
authors.forename: Y
authors.initials: Y
authors.lastname: Cao
authors.forename: L M
authors.initials: LM
authors.lastname: Rasmussen
authors.forename: M A
authors.initials: MA
authors.lastname: Karsdal
mesh_terms: D000328:Adult; D000818:Animals; D015415:Biomarkers / Q000097:blood / Q000378:metabolism; D024061:Collagen Type III / Q000097:blood* / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes / Q000097:blood* / Q000378:metabolism; D005109:Extracellular Matrix; D005260:Female; D006801:Humans; D008103:Liver Cirrhosis / Q000097:blood* / Q000378:metabolism; D008297:Male; D020780:Matrix Metalloproteinase 9 / Q000097:blood* / Q000378:metabolism; D008875:Middle Aged; D008954:Models, Biological; D051381:Rats; D017207:Rats, Sprague-Dawley; D012016:Reference Values; D055815:Young Adult
chemical_list: D015415:Biomarkers; D024061:Collagen Type III; D000939:Epitopes; D020780:Matrix Metalloproteinase 9