Fachbereich Veterinärmedizin



    Herstellung und Charakterisierung monoklonaler Antikörper gegen Neospora caninum (2002)

    Rehberg, Petra
    — 135 Seiten
    Institut für Parasitologie und Tropenveterinärmedizin

    Robert-von-Ostertag-Str. 7-13
    Gebäude 35, 22, 23
    14163 Berlin
    Tel.+49 30 838 62310 Fax.+49 30 838 62323

    Abstract / Zusammenfassung

    To gather more information about Neospora caninum antigenetic properties and to improve diagnostics, monoclonal antibodies against this parasite were produced and characterized. The results presented in this research are as follows:1 . The successful establishment of eight stable hybridoma cell-cultures out of wo fusions. Seven more monoclonal antibodies of previous fusion experiments at he BFAV were used for comparison.2. Seven of the monoclonal antibodies investigated did not recognize reduced antigen with the immunoblot method. Two antibodies failed to recognize their epitope after oxidation with sodium-meta-periodate. These reactions unveiled conformation epitopes and carbohydrate structures of the respective antigens.3. Utilizing several IFAT-methods as well immune precipitation of marked surface proteins of N. caninum tachyzoites the localisation of antigens detected by 13 of the 15 monoclonal antibodies investigated was recognised or confirmed. The results show that three antibodies recognize microneme-antigen, five antibodies are directed against dense granuleantigens and five of the antibodies investigated bind to the parasite surface.4. Six monoclonal antibodies reacted with excreted-secreted proteins of N. caninum. Three of these antibodies most likely recognize also surface antigens. This proves that parts of the parasite"s surface can also be used in excreted-secreted proteins.5. Only three of he monoclonal antibodies investigated cross-reacted with Toxoplasma gondii antigen in the immunoblot.6. The greatly reduced reactions of one antibody with the N. caninum isolate Nc-LivC 1 in the immunoblot suggest differences in the antigen patterns between different isolates. Another antibody exhibited mild variations of intensity in its reactivity with the detected antigen. Apart from these variations, no differences in antigenicity of a total of seven N. caninum isolates investigated were found in the immunoblot.7. The results of immunohistological tests suggest the detection of antigens specific to different stages in the lifecycle of N. caninum by the monoclonal antibodies investigated.Keywords: monoclonal antibodies, antibody, production, characterisation, Neospora caninum, neosporose, parasitic protozoa, dog, cattle.