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

Clin Biochem. 2010 Jul;43(10-11):899-904. doi: 10.1016/j.clinbiochem.2010.03.012. Epub 2010 Apr 7.


Objectives: 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.

Design and 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).

Conclusion: We have developed a novel ELISA for measuring a specific fragment CO3 generated by MMP-9 important in pathogenesis of liver fibrosis.

MeSH terms

  • Adult
  • Animals
  • Biomarkers / blood
  • Biomarkers / metabolism
  • Collagen Type III / blood*
  • Collagen Type III / metabolism
  • Enzyme-Linked Immunosorbent Assay / methods*
  • Epitopes / blood*
  • Epitopes / metabolism
  • Extracellular Matrix
  • Female
  • Humans
  • Liver Cirrhosis / blood*
  • Liver Cirrhosis / metabolism
  • Male
  • Matrix Metalloproteinase 9 / blood*
  • Matrix Metalloproteinase 9 / metabolism
  • Middle Aged
  • Models, Biological*
  • Rats
  • Rats, Sprague-Dawley
  • Reference Values
  • Young Adult


  • Biomarkers
  • Collagen Type III
  • Epitopes
  • Matrix Metalloproteinase 9