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Use of synthetic antibodies in immunohistochemistry
Tomečková, Kristýna ; Nejdl,, Lukáš (referee) ; Bezděková,, Jaroslava (advisor)
This diploma thesis deals with the optimization of the preparation of molecularly imprinted (MIP) nanoparticles doped with metal ions, selective for the selected protein. In this work, the model protein – chymotrypsinogen A was chosen as a template. The free radical polymerization method was used for the preparation of molecularly imprinted nanoparticles. Dopamine was used as a functional monomer because it is able to bind metal ions to each other. It also undergoes very rapid oxidative polymerization under alkaline conditions without the need for the addition of polymer reaction initiators. MIP optimization was performed by capillary electrophoresis with UV-Vis detection. The MIP thus prepared can serve as recognition elements in immunohistochemical analyzes that use LA-ICP-MS as a detection method. Their applicability for immunohistochemistry was studied using the dot block method.
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Molecularly imprinted polymers as recognition elements for the determination of disease markers
Vodová, Milada ; Vaculovičová,, Markéta (referee) ; Bezděková,, Jaroslava (advisor)
The diploma thesis is focused on the preparation and optimization of molecularly imprinted polymers (MIP) selective for chymotrypsinogen A as well as on the use of these MIP as recognition entities in the sensor. MIP was prepared by suspension polymerization using a mixture of functional methacrylate-based monomers. Prepared MIP was optimized (e.g. binding properties, selectivity and isolation of chymotrypsinogen from a complex matrix of human breath) by capillary electrophoresis with fluorescence detection ( = 532 nm). Finally, the combination of MIP with quartz microbalances was demonstrated as a promising sensor for the detection of proteins from human breath condensate.
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Molecularly imprinted polymers as a tool for the isolation of key biologically active molecules
Vodová, Milada ; Jaroslava, Bezděková (referee) ; Nejdl, Lukáš (advisor)
This bachelor thesis is focused on preparation and optimization of molecularly imprinted polymers that are selective for the selected nucleobase(uracil). Molecularly imprinted polymers, which wereused for selective uracil isolation, have been prepared non-covalent imprinting technique. As a function monomer was used in this case dopamine. The detection ofisolated uracil was made by capillary electrophoresis with absorption detection (–260 nm). Conditions for preparation of molecularly imprinted polymers,their binding properties, adsorption kinetics, and selectivity wereinvestigatedin detail. Mentioned polymer materials were as well used for uracil selective isolation and detection from complexsamples. Tomato products were used as complex samples. Uracil in those samples was usedasanindicator ofbacterial contamination.
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Use of synthetic antibodies in immunohistochemistry
Tomečková, Kristýna ; Nejdl,, Lukáš (referee) ; Bezděková,, Jaroslava (advisor)
This diploma thesis deals with the optimization of the preparation of molecularly imprinted (MIP) nanoparticles doped with metal ions, selective for the selected protein. In this work, the model protein – chymotrypsinogen A was chosen as a template. The free radical polymerization method was used for the preparation of molecularly imprinted nanoparticles. Dopamine was used as a functional monomer because it is able to bind metal ions to each other. It also undergoes very rapid oxidative polymerization under alkaline conditions without the need for the addition of polymer reaction initiators. MIP optimization was performed by capillary electrophoresis with UV-Vis detection. The MIP thus prepared can serve as recognition elements in immunohistochemical analyzes that use LA-ICP-MS as a detection method. Their applicability for immunohistochemistry was studied using the dot block method.
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Molecularly imprinted polymers as recognition elements for the determination of disease markers
Vodová, Milada ; Vaculovičová,, Markéta (referee) ; Bezděková,, Jaroslava (advisor)
The diploma thesis is focused on the preparation and optimization of molecularly imprinted polymers (MIP) selective for chymotrypsinogen A as well as on the use of these MIP as recognition entities in the sensor. MIP was prepared by suspension polymerization using a mixture of functional methacrylate-based monomers. Prepared MIP was optimized (e.g. binding properties, selectivity and isolation of chymotrypsinogen from a complex matrix of human breath) by capillary electrophoresis with fluorescence detection ( = 532 nm). Finally, the combination of MIP with quartz microbalances was demonstrated as a promising sensor for the detection of proteins from human breath condensate.
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Molecularly imprinted polymers as a tool for the isolation of key biologically active molecules
Vodová, Milada ; Jaroslava, Bezděková (referee) ; Nejdl, Lukáš (advisor)
This bachelor thesis is focused on preparation and optimization of molecularly imprinted polymers that are selective for the selected nucleobase(uracil). Molecularly imprinted polymers, which wereused for selective uracil isolation, have been prepared non-covalent imprinting technique. As a function monomer was used in this case dopamine. The detection ofisolated uracil was made by capillary electrophoresis with absorption detection (–260 nm). Conditions for preparation of molecularly imprinted polymers,their binding properties, adsorption kinetics, and selectivity wereinvestigatedin detail. Mentioned polymer materials were as well used for uracil selective isolation and detection from complexsamples. Tomato products were used as complex samples. Uracil in those samples was usedasanindicator ofbacterial contamination.
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