Monoclonal antibodies specific for bovine CD18
2001
Dusinsky, R. | Tomaskova, J. | Horovska, L. | Simon, M. (Slovenska Akademia Vied, Ivanka pri Dunaji (Slovak Republic). Ustav Biochemie a Genetiky Zivocichov)
Hybridoma cell lines producing monoclonal antibodies (mAbs) were obtained after two different immunizations of BALB/c mice with bovine lymphocytes using standard procedures for fusion of splenocytes with Sp2/0 myeloma cells, selections and cloning. Cytofluorometric analysis of bovine peripheral blood cells revealed that antibodies IVA35 and IVA218 gave positive reactions with 98-100 % lymphocytes, monocytes and granulocytes. The reactivity of both mAbs was also characterized by indirect immunoperoxidase staining of serial sections from healthy cattle tissues. Both mAbs immunoprecipitated proteins with apparent molecular weights of 170, 150 and 95 kDa, corresponding to CD11b, CD11c and CD18 molecules, from lysates of detergent-solubilized, surface sulphobiotin-labelled granulocytes. To determine a target subunit, both mAbs were further assayed for their capacity to immunoprecipitate alfa or beta subunits from the lysate that had been treated at pH 11.5 for 30 min. Epitopes for both mAbs were found to be present on the beta subunit. Our results demonstrate that IVA35 as well as IVA218 specifically recognize the CD18 molecule.
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