Abstract In order to define equine immunoglobulins (Igs) and to produce monoclonal reference Igs we fused equine peripheral blood mononuclear cells with X63-Ag8.653 non Ig producing murine myeloma cells. A total of 29 equine Ig producing equi-murine heterohybridomas were obtained, of which ten expressed equine Ig for more than 3 months. One of these heterohybridoma lines produced monoclonal IgM, an equine isotype which has not been available in monoclonal form before. Four lines secreted equine IgG of two distinct Ig heavy chain types as assessed by the molecular weight (MW), while the remaining five lines expressed only equine Ig light chains. A sixth Ig light chain expressing variant was obtained by cloning of one of the IgG producing heterohybridoma lines. These monoclonal IgGs were compared with previously described equine IgG monoclonal antibodies (mabs) by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). According to their MW we identified five groups of γ-chains and four groups of Ig light chains.