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Metody dekontaminace rekombinantních proteinů od bakteriálního lipopolysacharidu
CHARVÁTOVÁ, Lucie
In this work, three decontamination methods based on two-phase micellar system and affinity chromatography were used to decontaminate recombinant proteins from bacterial lipopolysaccharide and to determine which method is the most effective. The efficiency of this method was measured using various recombinant proteins at several protein concentrations. Three different assays, two chromogenic and one fluorogenic, were used to measure the concentration of endotoxins in samples. The most accurate method for measuring the concentration of endotoxins was determined.
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Construction of a genetically detoxified Bordetella pertussis strain to develope a new generation of whole-cell vaccine
Bočková, Barbora ; Holubová, Jana (advisor) ; Seydlová, Gabriela (referee)
Bordetella pertussis is a strictly human pathogen colonizing the upper respiratory tract, causing a respiratory disease known as whooping cough or pertussis. The introduction of whole-cell vaccines and acellular vaccines, resulted in a significant reduction in the incidence of disease and reduce the fatalities associated with infection. However, epidemiological data show a significant increase in the incidence of the disease in recent decades. The increasing incidence is mainly attributed to the transition from the whole- cell vaccine to an acellular vaccine. Based on research from recent years has shown that acellular vaccines have many drawbacks, and it is therefore necessary to change the vaccination strategy. One possible solution to the situation is the development of a new generation of whole-cell vaccines with reduced reactogenicity. The new whole-cell vaccine was prepared by a genetically modified B. pertussis strain. B. pertussis was modified using allelic exchange to develop strain encoding enzymatically inactive pertussis toxin, modified lipid A and lacking dermonecrotic toxin. This combination of genetic modifications in mice led to a decrease in reactogenicity test vaccine in vivo. In case of intranasal infection whole-cell vaccine containing genetically modified strain is providing...
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Construction of a genetically detoxified Bordetella pertussis strain to develope a new generation of whole-cell vaccine
Bočková, Barbora ; Holubová, Jana (advisor) ; Seydlová, Gabriela (referee)
Bordetella pertussis is a strictly human pathogen colonizing the upper respiratory tract, causing a respiratory disease known as whooping cough or pertussis. The introduction of whole-cell vaccines and acellular vaccines, resulted in a significant reduction in the incidence of disease and reduce the fatalities associated with infection. However, epidemiological data show a significant increase in the incidence of the disease in recent decades. The increasing incidence is mainly attributed to the transition from the whole- cell vaccine to an acellular vaccine. Based on research from recent years has shown that acellular vaccines have many drawbacks, and it is therefore necessary to change the vaccination strategy. One possible solution to the situation is the development of a new generation of whole-cell vaccines with reduced reactogenicity. The new whole-cell vaccine was prepared by a genetically modified B. pertussis strain. B. pertussis was modified using allelic exchange to develop strain encoding enzymatically inactive pertussis toxin, modified lipid A and lacking dermonecrotic toxin. This combination of genetic modifications in mice led to a decrease in reactogenicity test vaccine in vivo. In case of intranasal infection whole-cell vaccine containing genetically modified strain is providing...
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