Reference : Development of an enzyme-linked immunosorbent assay for equine neutrophil elastase measu...
Scientific journals : Article
Life sciences : Biochemistry, biophysics & molecular biology
Life sciences : Veterinary medicine & animal health
http://hdl.handle.net/2268/1342
Development of an enzyme-linked immunosorbent assay for equine neutrophil elastase measurement in blood: Preliminary application to colic cases.
English
de la Rebière de Pouyade, Geoffroy mailto [Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux >]
Franck, Thierry mailto [Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux >]
Salciccia, Alexandra mailto [Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Département clinique des animaux de compagnie et des équidés >]
Deby-Dupont, Ginette mailto [> > > >]
Grulke, Sigrid mailto [Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Département clinique des animaux de compagnie et des équidés >]
Vander Heyden, Laurent mailto [Université de Liège - ULg > > Clinique des grands animaux (chirurgie) >]
Sandersen, Charlotte mailto [Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Département clinique des animaux de compagnie et des équidés >]
Serteyn, Didier mailto [Université de Liège - ULg > Département clinique des animaux de compagnie et des équidés > Anesthésiologie gén. et pathologie chirurg. des grds animaux >]
2010
Veterinary immunology and immunopathology
Yes
International
1873-2534
1873-2534
[en] Equine neutrophil elastase (NE) is a protease released in inflammatory diseases and participating in tissue destruction. To measure NE in horse plasma to assess its role in pathological conditions, we purified elastase from equine neutrophils by a double step chromatography and obtained a pure protein of 27kDa, 4kDa smaller than the NE 2A previously purified (Scudamore et al., 1993; Dagleish et al., 1999), which was likely to be NE 2B. We developed an ELISA by using two specific polyclonal antibodies obtained from rabbit and guinea pig. The sandwich complex was detected using a secondary antibody conjugated to alkaline phosphatase. The ELISA showed good precision and accuracy, with intra- and inter-assay coefficients of variation below 10% for equine NE concentrations ranging from 1.875 to 60ng/ml. A stable plasma NE value, unaffected by the delay of centrifugation (over 4h), was obtained with plasma from EDTA anticoagulated blood. The mean value (+/-SEM) measured in 37 healthy horses was 32.53+/-4.6ng/ml. NE level in plasma of horses with colic at the time of admission was significantly higher than in healthy horses. Our results indicate that the ELISA technique we developed to measure plasmatic NE is a powerful tool for studying the role of elastase in equine inflammatory disease. In future, the application will be extended to other equine biological fluids.
Centre de l’Oxygène, Recherche et Développement - CORD
Fonds pour la formation à la Recherche dans l'Industrie et dans l'Agriculture (Communauté française de Belgique) - FRIA
Researchers ; Professionals ; Students
http://hdl.handle.net/2268/1342
10.1016/j.vetimm.2009.10.023

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