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Development of new types of biocompatible hemodialysis membranes for separation of biomolecules
Kohlová, Michaela ; Solich, Petr (advisor) ; Kujovská Krčmová, Lenka (referee) ; Pena, Angelina (referee)
Charles University, Faculty of Pharmacy in Hradec Králové / University of Porto, Faculty of Pharmacy Department of Analytical Chemistry / Department of Chemical Sciences Candidate: Mgr. Michaela Kohlová Supervisor: prof. RNDr. Petr Solich, CSc. Consultants: prof. Maria de Conceição Branco Montenegro assoc. prof. Alice Santos-Silva Title of dissertation thesis: Development of new types of biocompatible haemodialysis membranes for separation of biomolecules This dissertation thesis was focused on the development, preparation, and optimization of flat sheet polysulfone membranes, which would meet the characteristics required for membranes used for haemodialysis treatment. The thin porous membranes were prepared by spin coating method, followed by phase inversion via immersion precipitation. The composition, as well as the preparation process of the membranes, were optimized, to obtain a membrane with good mechanical performance in flow conditions, adequate selective separation ability, and biocompatibility. The prepared membranes with the appropriate characteristics were modified with bioactive compounds to minimize oxidative stress and/or inflammation, which are common complications in haemodialysis treated patients. Three different modification approaches were used to prepare the bioactive...
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Cloning, expression and purification of Francisella tularensis disulfide oxidoreductase
Kohlová, Michaela ; Šimůnek, Tomáš (advisor) ; Zemanová, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michaela Kohlová Supervizor: doc. PharmDr. Tomáš Šimůnek Supervizor - konsultant: Mgr. Monika Schmidt Title of diploma thesis: Cloning, expression and purification of Francisella tularensis disulfide oxidoreductase Conserved hypothetical lipoprotein called FTT1103 (for Francisella tularensis subsp. tularensis) is one of the virulence factors of gramnegative intracellular bacteria Francisella tularensis. This protein is homologue to protein E. coli Dsba. Periplasmatic protein DsbA is member of disulfide oxidoreductase family and it is responsible for disulfide bond formation in newly secreted proteins. The disulfide bonds are essential for right conformation and activity in the proteins. Biotransformatic analysis of protein FTT1103 showed, that this protein contains N-terminal FKBP domain. The domain has probably dimerization activity and it is expected to have chaperone activity too. The aim of the presented diploma thesis was to verify, if the domain FKBP_N is really responsible for correct function and activity of DsbA protein. Using molecular biology methods we prepared mutant gene with deleted domain FKBP_N (dsbAΔFKBP_N). We cloned this gene into plasmidplazmid E. coli pET28b,...
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Development of new types of biocompatible hemodialysis membranes for separation of biomolecules
Kohlová, Michaela ; Solich, Petr (advisor) ; Kujovská Krčmová, Lenka (referee) ; Pena, Angelina (referee)
Charles University, Faculty of Pharmacy in Hradec Králové / University of Porto, Faculty of Pharmacy Department of Analytical Chemistry / Department of Chemical Sciences Candidate: Mgr. Michaela Kohlová Supervisor: prof. RNDr. Petr Solich, CSc. Consultants: prof. Maria de Conceição Branco Montenegro assoc. prof. Alice Santos-Silva Title of dissertation thesis: Development of new types of biocompatible haemodialysis membranes for separation of biomolecules This dissertation thesis was focused on the development, preparation, and optimization of flat sheet polysulfone membranes, which would meet the characteristics required for membranes used for haemodialysis treatment. The thin porous membranes were prepared by spin coating method, followed by phase inversion via immersion precipitation. The composition, as well as the preparation process of the membranes, were optimized, to obtain a membrane with good mechanical performance in flow conditions, adequate selective separation ability, and biocompatibility. The prepared membranes with the appropriate characteristics were modified with bioactive compounds to minimize oxidative stress and/or inflammation, which are common complications in haemodialysis treated patients. Three different modification approaches were used to prepare the bioactive...
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Development of new types of biocompatible hemodialysis membranes for separation of biomolecules
Kohlová, Michaela ; Solich, Petr (advisor) ; Kujovská Krčmová, Lenka (referee) ; Pena, Angelina (referee)
Charles University, Faculty of Pharmacy in Hradec Králové / University of Porto, Faculty of Pharmacy Department of Analytical Chemistry / Department of Chemical Sciences Candidate: Mgr. Michaela Kohlová Supervisor: prof. RNDr. Petr Solich, CSc. Consultants: prof. Maria de Conceição Branco Montenegro assoc. prof. Alice Santos-Silva Title of dissertation thesis: Development of new types of biocompatible haemodialysis membranes for separation of biomolecules This dissertation thesis was focused on the development, preparation, and optimization of flat sheet polysulfone membranes, which would meet the characteristics required for membranes used for haemodialysis treatment. The thin porous membranes were prepared by spin coating method, followed by phase inversion via immersion precipitation. The composition, as well as the preparation process of the membranes, were optimized, to obtain a membrane with good mechanical performance in flow conditions, adequate selective separation ability, and biocompatibility. The prepared membranes with the appropriate characteristics were modified with bioactive compounds to minimize oxidative stress and/or inflammation, which are common complications in haemodialysis treated patients. Three different modification approaches were used to prepare the bioactive...
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Cloning, expression and purification of Francisella tularensis disulfide oxidoreductase
Kohlová, Michaela ; Šimůnek, Tomáš (advisor) ; Zemanová, Lucie (referee)
Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Michaela Kohlová Supervizor: doc. PharmDr. Tomáš Šimůnek Supervizor - konsultant: Mgr. Monika Schmidt Title of diploma thesis: Cloning, expression and purification of Francisella tularensis disulfide oxidoreductase Conserved hypothetical lipoprotein called FTT1103 (for Francisella tularensis subsp. tularensis) is one of the virulence factors of gramnegative intracellular bacteria Francisella tularensis. This protein is homologue to protein E. coli Dsba. Periplasmatic protein DsbA is member of disulfide oxidoreductase family and it is responsible for disulfide bond formation in newly secreted proteins. The disulfide bonds are essential for right conformation and activity in the proteins. Biotransformatic analysis of protein FTT1103 showed, that this protein contains N-terminal FKBP domain. The domain has probably dimerization activity and it is expected to have chaperone activity too. The aim of the presented diploma thesis was to verify, if the domain FKBP_N is really responsible for correct function and activity of DsbA protein. Using molecular biology methods we prepared mutant gene with deleted domain FKBP_N (dsbAΔFKBP_N). We cloned this gene into plasmidplazmid E. coli pET28b,...
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