National Institute of Animal Health (NIAH)

Topics in Animal Health Research 2012

18. Establishment of a quantitative bovine CXCL8 sandwich ELISA with a newly developed monoclonal antibody


Three IgG class anti-bovine CXCL8 (bCXCL8) monoclonal antibody (mAb)-secreting hybridomas, namely, SH8-8D7, SH8-12A5 and SH8-2A1, were developed. All three mAbs detected recombinant bCXCL8 (rbCXCL8) by immunoprecipitation and western blotting. SH8-2A1 neutralised the chemotactic activity of rbCXCL8 towards neutrophils. The ELISA quantified bCXCL8 by the combination of SH8-12A5 and biotin-SH8-2A1. The detection range was 20-1000 pg/ml. This sandwich ELISA detected native bCXCL8 derived from the cultured supernatant of mononuclear cells from bovine peripheral blood that were stimulated with ConA, LPS, or PHA. Furthermore, SH8-2A1 detected bCXCL8 in formalin-fixed, paraffin-embedded pneumonic calf tissue. These findings indicate that this newly developed anti-bCXCL8 mAbs have the potential to contribute to research on bovine inflammatory responses and immunology.
(Center for Animal Disease Control and Prevention)


Hirota J. et al. (2011) European Cytokine Network 22(1):73-80