Serological differentiation of the potato-cyst nematodes Globodera pallida and G. rostochiensis. II. Preparation and characterization of species-specific monoclonal antibodies

Hybridomas producing antibodies which react with thermostable protein antigens isolated from the potato cyst nematode species Globodera rostochiensis (TSRoP) and G. pallida (TSPaP) were isolated. Three of the isolated hybridomas (WGP 1, WGP 2 and WGP 3) produce antibodies that react with preferent affinity with protein antigens isolated from G. pallida, and two (WGR 11 and WGR 12) produce antibodies which bind preferentially to G. rostochiensis. Binding constants were determined to quantitate the differences in affinity of WGP 1, WGP 2, WGP 3, WGR 11 and WGR 12 for the protein antigens from both nematode species, and to asses the similarity in affinity for either protein antigen with respect to the other non-specific antibodies. In immunoblotting experiments a binding could be demonstrated, for most antibodies, to two thermostable proteins with apparent molecular weights of 20.6/20.8 kD for G. rostochiensis and 20.5/21.0 kD for G. pallida. The reactivity of the monoclonal antibodies with thermostable protein antigens from other common cyst nematodes was also investigated. All monoclonal antibodies, which are not specific for TSRoP or TSPaP, bind to thermostable proteins of these cyst nematode species. The use of some of the isolated monoclonal antibodies to improve the diagnosis of potato cyst nematodes in soil samples is discussed.