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Interaction of influenza virus with cell defence mechanisms
Vochyánová, Klára ; Drda Morávková, Alena (advisor) ; Mikušová, Gabriela (referee)
Influenza virus infection is one of the most current problems nowadays. Its unique ability to strongly inhibit cell immune response on many different levels and its pandemic potential make it a subject of interest of many research groups. The Influenza virus uses mainly NS1 protein to inhibit the immune response. NS1 protein is able, on one hand, to bind RNA and mask it against recognition by cellular sensors and other proteins. On the other hand, NS1 protein possesses a catalytic domain. Using this domain it interacts with many cellular proteins, interferes with signal transduction and guarantees successful infection. NS1 protein is one of principal pathogenicity instruments of the Influenza virus and it deserves appropriate attention.
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Preparation of Monoclonal Antibodies Against VP2 Protein of Human Polyomaviruses
Vochyánová, Klára ; Drda Morávková, Alena (advisor) ; Růžičková, Šárka (referee)
Aim of this diploma thesis was to prepare two protein antigens and two monoclonal antibodies, all based on VP2 minor protein of human polyomaviruses BK virus and Merkel Cell Polyomavirus. One monoclonal antibody was being prepared against unique part of VP2 protein (N-terminal epitope, not present in VP3 protein). A cell line producing such monoclonal antibody has never been established before due to low immunogenicity of the epitope. Our approach was successful in terms of mouse immunization, however, serious problems with hybridoma line stability appeared later during the preparation process. Preparation of antibody targeted to the sequence of VP2 protein of Merkel Cell Polyomavirus was another aim of this thesis. Mouse immunization and hybridoma fusion were performed successfully. After four rounds of cloning in order to purify an established clone, nine clones were cultivated in larger scale. This cultivation probably led to diminished antibody specificity and loss of production ability in most of the hybridoma cells. One more cloning should give rise to an established clone with sufficient production. Two preparations of protein antigens were performed in two expression systems. DNA encoding C-terminally truncated protein VP2 of BK virus fused with His-tag was cloned into a vector suitable for...
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Interaction of influenza virus with cell defence mechanisms
Vochyánová, Klára ; Drda Morávková, Alena (advisor) ; Mikušová, Gabriela (referee)
Influenza virus infection is one of the most current problems nowadays. Its unique ability to strongly inhibit cell immune response on many different levels and its pandemic potential make it a subject of interest of many research groups. The Influenza virus uses mainly NS1 protein to inhibit the immune response. NS1 protein is able, on one hand, to bind RNA and mask it against recognition by cellular sensors and other proteins. On the other hand, NS1 protein possesses a catalytic domain. Using this domain it interacts with many cellular proteins, interferes with signal transduction and guarantees successful infection. NS1 protein is one of principal pathogenicity instruments of the Influenza virus and it deserves appropriate attention.
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