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