PREPARATION AND CHARACTERIZATION OF MONOCLONAL-ANTIBODIES AGAINST BOVINE MYELOPEROXIDASE

We established eight cloned B-cell hybridomas producing monoclonal ant ibodies (Mo abs) against bovine myeloperoxidase (MPO). These anti-MPO (AM) Mo abs, designated AM(1)-AM(8), all reacted similarly to three ch romatographic forms of MPO, isolated from a single donor, in an enzyme linked immunosorbent assay. According to immunoblot analysis and ELIS A the AM Mo abs are specific to bovine MPO and show no cross reactivit y with other neutrophil granule proteins such as lactoferrin, lactoper oxidase and serum albumin. In immunoblot analyses IgG(1) class AM(1), AM(2), AM(3) and AM(4) Mo abs immunostained the heavy subunit of the M PO (57 kDa). Additionally, the AM Mo abs seem to bind either the react ive site or epitopes on bovine MPO that affect the peroxidase activity of this enzyme. AM Mo abs reacted specifically with neutrophils but d id not react with lymphocytes or epithelial cells. The present study s hows that these AM Mo abs could be used for developing immunoassays to measure bovine MPO from biological fluids and for localizing neutroph ils at sites of infections. They could also be useful in studies asses sing the involvement of MPO in inflammatory processes in bovine specie s.