title: Development and validation of an enzyme-linked immunosorbent assay for the quantification of a specific MMP-9 mediated degradation fragment of type III collagen--A novel biomarker of atherosclerotic plaque remodeling.
abstract: OBJECTIVE
  Degradation of collagen in the arterial wall by matrix metalloproteinases is the hallmark of atherosclerosis. We have developed an ELISA for the quantification of type III collagen degradation mediated by MMP-9 in urine.
  
  
  METHODS
  A monoclonal antibody targeting a specific MMP-9 generated fragment of collagen III was used in a competitive ELISA. The assay was validated in urine and arterial tissue of Apolipoprotein-E knockout (ApoE-KO) mice.
  
  
  RESULTS
  The lower limit of detection was 0.5ng/mL, intra- and inter-assay coefficients of variation were below 10%. By the end of 20weeks of the study, urine levels of the novel CO3-610 biomarker in ApoE-KO mice increased by two-fold (p<0.0001) and were three-fold higher than in control mice. Western blots confirmed high expression of CO3-610 in arterial extracts of ApoE-KO mice.
  
  
  CONCLUSIONS
  We have developed a novel competitive ELISA, capable of measuring a urine biomarker indicative of pathological extracellular matrix remodeling in a mouse model of atherosclerosis.
authors.affiliation: Nordic Bioscience A/S, Herlev, Denmark; University of Southern Denmark, Odense, Denmark. nb@nordicbioscience.com
authors.forename: Natasha
authors.initials: N
authors.lastname: Barascuk
authors.forename: Efstathios
authors.initials: E
authors.lastname: Vassiliadis
authors.forename: Lise
authors.initials: L
authors.lastname: Larsen
authors.forename: Jianxia
authors.initials: J
authors.lastname: Wang
authors.forename: Qinlong
authors.initials: Q
authors.lastname: Zheng
authors.forename: Rui
authors.initials: R
authors.lastname: Xing
authors.forename: Yu
authors.initials: Y
authors.lastname: Cao
authors.forename: Christine
authors.initials: C
authors.lastname: Crespo
authors.forename: Isabelle
authors.initials: I
authors.lastname: Lapret
authors.forename: Massimo
authors.initials: M
authors.lastname: Sabatini
authors.forename: Nicole
authors.initials: N
authors.lastname: Villeneuve
authors.forename: Jean-Paul
authors.initials: JP
authors.lastname: Vilaine
authors.forename: Lars Melholt
authors.initials: LM
authors.lastname: Rasmussen
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Register
authors.forename: Morten A
authors.initials: MA
authors.lastname: Karsdal
mesh_terms: D000818:Animals; D001057:Apolipoproteins E / Q000172:deficiency; D015415:Biomarkers / Q000652:urine; D002784:Cholesterol / Q000097:blood; D024061:Collagen Type III / Q000032:analysis* / Q000652:urine; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism*; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D010446:Peptide Fragments / Q000032:analysis* / Q000737:chemistry; D058226:Plaque, Atherosclerotic / Q000097:blood / Q000175:diagnosis* / Q000652:urine; D012372:ROC Curve; D012015:Reference Standards; D015203:Reproducibility of Results; D013379:Substrate Specificity; D014280:Triglycerides / Q000097:blood
chemical_list: D001057:Apolipoproteins E; D015415:Biomarkers; D024061:Collagen Type III; D010446:Peptide Fragments; D014280:Triglycerides; D002784:Cholesterol; D020780:Matrix Metalloproteinase 9