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.
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.
Barascuk
N
N
Nordic Bioscience A/S, Herlev, Denmark. nb@nordicbioscience.com
Veidal
S S
SS
Larsen
L
L
Larsen
D V
DV
Larsen
M R
MR
Wang
J
J
Zheng
Q
Q
Xing
R
R
Cao
Y
Y
Rasmussen
L M
LM
Karsdal
M A
MA
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
D015415:Biomarkers; D024061:Collagen Type III; D000939:Epitopes; D020780:Matrix Metalloproteinase 9