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Preparation of monoclonal antibodies against Borrelia
ŠVEJDOVÁ, Veronika
In my bachelor thesis I deal with preparation of monoclonal antibodies against antigens of Borrelia. In theoretical section I describe genus Borrelia and antibodies. The introduction deals with the history of discovery of the spirochete that causes Lyme disease, vectors of this disease and classification of Borreliae according to geographical occurrence. Taxonomic categorization and description of the bacterium followers. It is focused on flagellum, arrangement of genetic material and on external surface antigens important for this bacterium. In the next section I describe Borrelia vector - host interactions and the development of immune response during the disease. Dermal symptoms of the Lyme disease are described in detail. A few words about relaxing fever are added. This chapter ends with description of laboratory diagnostics of Borrelia burgdorferi. Next, I focused on the characterization of antibodies and on preparation of monoclonal antibodies by hybridoma technology. I briefly describe ways of their utilization. Practical part of Bachelor thesis was carried out in the laboratories of Faculty of Science. Here, with the help of hybridoma technology, I fused myeloma cells with splenocytes of immunized mice producing hybridomas. These hybridomas I cultivated in medium, which I subsequently tested for occurrence of antibodies by ELISA. Hybridomas having high productivity were cloned, frozen and stored in liquid nitrogen. Finally I purified monoclonal antibodies, concentrated them and tested their titre.
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