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Preparation and characterization of surface-modified nanoliposomes using click chemistry techniques
Frydrychová, Aneta ; Bartoš,, Milan (referee) ; Turánek, Jaroslav (advisor)
Over the past decades the liposomes have been intensively studied for their unique properties which predispose them for use as drug delivery systems or for constructions of vaccines. This diploma thesis provides an overview of their most important properties, preparation options and their surface modification. The aim of this thesis is thus a preparation and characterization of the nanoliposomes and their surface modification. The liposomes were prepared by lipid film hydration method and mannan polysacharide was used for surface modification. Due to the use of a lipid with an N-oxy group, the modification was carried out via an oxime ligation via click chemistry. Nanoliposomes were characterized by series of physicochemical methods such as TEM, DLS, FT-IR or nano-flow cytometry. Part of the thesis is a study of the interactions of liposomes accomplished on selected cell lines to verify whether they stimulate immune response pathways. Its results confirmed activation of the NLRP3 inflammasome leading to the production of pro-inflammatory cytokines (IL-1). Thus, these polymer coated nanoliposomes are potentially useful as vaccine adjuvants.
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Studying protein interactions with expanded genetic code
Tekel, Andrej ; Obšil, Tomáš (advisor) ; Fuertes Vives, Gustavo (referee)
Nature, using proteins composed of approximately 20 amino acids repertoire, is able to perform a large number of admirable things, many of which we are still not quite able to mimic. However, it has to be said, that the set of chemical scaffolds available to the nature is somewhat limited. Therefore, it seems plausible to assume, that we could make things better simply by introducing novel scaffolds and exploring chemical space so far unavailable. It is exactly this idea upon which expanded genetic code is based on. To set ourselves free from limitations imposed by standard genetic code. This thesis will aim to provide an overview of various attempts to this problem and will try to do so in a distinct context of studying protein interactions. I will thus try to highlight methods by which we are currently able to expand genetic code, specific chemistries and physical properties of non-canonical amino acids and biophysical methods which can profit from genetic code expansion. Keywords: expanded genetic code, non-canonical amino acid, amber codon, fluores- cence, magnetic resonance, crosslinking, click chemistry
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Preparation and characterization of surface-modified nanoliposomes using click chemistry techniques
Frydrychová, Aneta ; Bartoš,, Milan (referee) ; Turánek, Jaroslav (advisor)
Over the past decades the liposomes have been intensively studied for their unique properties which predispose them for use as drug delivery systems or for constructions of vaccines. This diploma thesis provides an overview of their most important properties, preparation options and their surface modification. The aim of this thesis is thus a preparation and characterization of the nanoliposomes and their surface modification. The liposomes were prepared by lipid film hydration method and mannan polysacharide was used for surface modification. Due to the use of a lipid with an N-oxy group, the modification was carried out via an oxime ligation via click chemistry. Nanoliposomes were characterized by series of physicochemical methods such as TEM, DLS, FT-IR or nano-flow cytometry. Part of the thesis is a study of the interactions of liposomes accomplished on selected cell lines to verify whether they stimulate immune response pathways. Its results confirmed activation of the NLRP3 inflammasome leading to the production of pro-inflammatory cytokines (IL-1). Thus, these polymer coated nanoliposomes are potentially useful as vaccine adjuvants.
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Synthesis and characterization of new insulin analogs with a triazole bridge at the C-terminus of the B chain
Kuntová, Vendula ; Jiráček, Jiří (advisor) ; Ryšlavá, Helena (referee)
Insulin is a peptide hormone responsible for maintaining glucose homeostasis in the circulation. Insulin interacts with two isoforms of the insulin receptor, IR-A and IR-B, which have different tissue distribution. IR-A is supposed to have rather mitogenic function and IR-B rather metabolic function. The goal of this study was to develop insulin analog, which will be more selective for IR-B than human insulin. We prepared three new insulin analogs with a 1,2,3-triazole bridge at the positions B26 and B29. The triazole bridge was formed by Cu(I)- catalysed cycloaddition between side chains of azidopentanoic acid (N3Pent) at B26 and propargylglycine (Prg) at B29. The analogs differed in configurations on C carbons of unusual amino acids at the positions B26 and B29. Specifically, we prepared insulin analog 1 with D- N3PentB26 and D-PrgB29, insulin analog 2 with D-N3PentB26 and L-PrgB29 and insulin analog 3 with L-N3PentB26 and D-PrgB29. New analogs were tested for their binding to both isoforms (IR-A and IR-B) of the insulin receptor. Analogs 1 and 2 were less potent in binding than human insulin and had no selectivity for receptor isoforms. Analog 3 was 4-times more potent in binding to IR-B and 2-times more potent in binding to IR-A than human insulin. However, the binding selectivity of the...
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