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Biofyzikální studium malých RNA
Šmerková, Kristýna
Thanks to the prove of connection between the aberrant occurrence of small RNA and various diseases and their potential in diagnostics and treatment led to discovery of new methods and materials facilitating their detection and targeted transport during gene therapy. This work summarizes present knowledge about chosen groups of small RNA, their significance in medical science and the possibilities of their detection. This work primarily concentrates on combination of magnetic separation with electrochemical detection. Magnetic particles (MPs) with different surface modifications were used for isolation. Non-specific isolation was carried out using silanol-coated MPs; streptavidin-coated MPs modified with specific biotinylated probe were used for specific separation. Square wave voltammetry (SWV) was used as a very sensitive electrochemical detection method. Optimized method based on specific magnetic separation with SWV was able to reach nanomolar detection limit (4 nM) with microRNA. The method was applied on human embryonic cells for specific isolation and detection of miR-124. The CdTe quantum dots (QDs) were studied as a nanomaterial tool for nucleic acid detection. The QDs were modified with streptavidin for their bioconjugation with biotinylated molecules were used. Interaction of QDs with nucleic acids was studied using capillary electrophoresis.
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Biomolecules influence on optical properties of deposited quantum dots
Roček, Vojtěch ; Mravec, Filip (referee) ; Drbohlavová, Jana (advisor)
This thesis is focused on creation of quantum dots of titanium dioxide with help of nanoporous alumina mask and on funcionalization of their surface in purpose to increase the quantum yield of its. This thesis is devided into 6 main chapters, where the glossary topic of quantum dots creation process and ways of practical usage of these quantum dots are made. The theoretical knowledge is used for creation of titanium dioxide quantum dots, which was deposited by gold and funcionalized with glutation. The results are shown and discussed in the end of this work.
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Biosensors based on modified nanoparticles
Mihajlović, Ana ; Majzlíková, Petra (referee) ; Hubálek, Jaromír (advisor)
This thesis deals with biosensors based on modified semiconductor quantum dots(AIIBIII). This work is divided into three main parts. The first deals with the theory necessary for the use of QDs in bioapplications, there are described methods of synthesis, application and bioconjugation of QDs . In the practical part, GSH-CdTe, CdTe-MPA and TGA-CdTe quantum dots were prepared by direct colloidal synthesis. These quantum dots were then modified by EDC in order to increase affinity to BSA (bovine serumalbumin). For the analysis of samples were used Infinite M200,Tecan and HORIBA Jobin Yvon FluoroLog detector Quanta . The third part is the summary of results and work achievements.
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Biosensors based on modified TiO2 nanoparticles
Valkovičová, Jiřina ; Adam, Vojtěch (referee) ; Hubálek, Jaromír (advisor)
The work is focused on quantum dots (QDs) array fabrication from titanium dioxide using template based non-lithographic method and their luminescence (fluorescence) properties study. The paper describes TiO2 QDs surface modification with gold using suitable template technique and subsequent functionalization of gold modified QDs surface via oligonucleotides. This functinalization step provides a suitable base for subsequent hybridization reaction with detected nucleic acid. The system of biosensor (TiO2 QDs modified with gold and biomolecules) is designed for rapid optical diagnosis of viral diseases.
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Development of Instrumentation and Methodology in Proteomic and Environmental Analysis
Hezinová, Věra ; Bobáľová, Janette (referee) ; Lubal,, Přemysl (referee) ; Čáslavský, Josef (referee) ; Klepárník, Karel (advisor)
Tato práce je zaměřena jak na cílený tak na přehledný přístup ve studiu proteomiky. Cílená proteomika přináší informace o přítomnosti proteinu a jeho lokalizaci v buňce či tkáni pomocí luminiscenčních značek na bázi kvantových teček, zatímco přehledná proteomika se zabývá identifikací změn v proteomu dvou nebo více jedinců stejného druhu vystavených různým podmínkám. Protože proteomika vyžaduje vysoce citlivé separační a identifikační techniky, byly v této práci ověřeny různé metody zlepšení citlivosti kapilární elektroforézy s hmotnostní detekcí. Použití rozhraní s kapalinovým spojem pro spojení těchto dvou technik, které zajišťuje vyšší citlivost analýz, bylo také ověřeno analýzou metabolitů etanolu a kokainu v lidské moči. Zavedené techniky instrumentace jsou využitelné při posouzení vlivu významných faktorů životního prostředí na živé systémy jak na buněčné tak na molekulární úrovni.
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Plasmon enhanced photoluminescence
Édes, Zoltán ; Kůsová,, Kateřina (referee) ; Šikola, Tomáš (advisor)
Diplomová práce se zabývá fotoluminiscencí polovodičových materiálů zesílené plazmonovými polaritony. Je popsána základní teorie interakce mezi lokalizovanými povrchovými plazmonovými polaritony a fotoluminiscenčními látkami. Dva mechanismy, které mohou vést k fotoluminiscenci zesílené plazmonovými polaritony jsou diskutovány. Následně je popsán návrh aparatury pro měření fotoluminiscence a způsob její realizace. Funkčnost aparatury je ověřena měřením fotoluminiscenčních spekter objemového GaN, nanokrystalického Si a CdTe kvantových teček. Nakonec je zkoumána metoda přípravy vzorků sestávajících z kovových nanokuliček a fotoluminiscenčně aktívních CdTe kvantových teček.
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Study of influence of quantum dots to biological systmes and their components
Koudelková, Zuzana ; Zítka, Ondřej (referee) ; Provazník, Ivo (advisor)
The aim of this thesis is to summarize the available evidence about quantum dots and their effects on living systems. The text describes methods for the preparation of quantum dots with respect to their characteristics (size, fluorescence wavelength) and methods of quantum dots bio-functionalization of biomolecules. In living organisms is a large number of proteins, because these are considered as one of the main components of the interaction of organisms. Therefore, the work also provides basic information about proteins. Finally, there are described various methods by which the quantum dots may be characterized mainly by differential pulse voltammetry measurement zeta potential and fluorescence measurement. The main objective of this work is to propose models of different environments in which will be degradation of quantum dots with regard to the evaluation of acquired kinetic parameters for predicting the stability of individual quantum dots.
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Synthesis of core/shell quantum dots for diagnostics
Mihajlović, Ana ; Vaculovičová, Markéta (referee) ; Pekárková, Jana (advisor)
This thesis deals with biosensors based on modified semiconductor core/shell quantum dots (QDs) for diagnosis. The work is divided into four main parts. The first one discusses the theory required for the use of QDs in bioaplications, there are described methods of synthesis, modification, application and bioconjugation of QDs. In the experimental part, CdTe/ZnS QDs with core/shell structure were prepared, in which the core was modified by MPA, GSH and TGA. In the next step, these QDs were further modified using CDI, EDC and NHS as mediators in order to increase affinity to BSA (bovine serum albumine) and IgG (imunoglobuline G). Prepared conjugates were characterized by fluorescence spectroscopy (Infinite M200Pro, Tecan) and capillary electrophoresis (Agilent 7100).
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Diagnostické biosenzory pro encefalopatie způsobené priony
Šobrová, Pavlína
Prion diseases are fatal transmisible neurodegenerative and infectious disorders (TSEs) of humans and animals, characterized by structural transition of the host-encoded cellular prion protein (PrPC) into the aberrantly folded pathologic isoform PrPSc. The main aim of this work is to summarize present information about prion diseases and their possibilities of determination pointed to electrochemical techniques. For this purpose cyclic voltammetry (CV), differential pulse voltammetry, differential pulse voltammetry Brdicka reaction and chronopotentiometric stripping analysis (CPSA) were used. The estimated detection limits were 32 ug/ml by CV, 16 ug/ml by DPV, 16 ug/ml by DPV -- Brdicka reaction and 8 ug/ml by CPSA. Subsequently, the influence of heat denaturation was observed. It clearly follows from the obtained results that signals of prion decreased linearly depending on the duration of the heat treatment at 99°C for various time intervals 0, 15, 30, 45, and 60 min. Moreover, we aimed our attention on studying of prion protein interaction with CdTe quantum dots (QDs) using electrochemistry. Primarily, we characterized electrochemical properties of QDs and the detection limit at 100 fg/ml was estimated. Further, electrochemical study of prion and QD interactions was carried out to find the most suitable conditions for sensitive detection of prion proteins. Detection limit (3 S/N) was estimated as 1 fg in 5 ul. This makes labeling of proteins with QDs of great importance due to easy applicability and possibility to use in miniaturized devices, which can be used in situ. This should open new possibilities how to determine the presence of these proteins on surgical equipment and other types of materials, which could be contagious.
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Modifikované peptidy ve spojení s kvantovými tečkami
Janů, Libor
In this study, water soluble CdTe QDs of different emission wavelengths were prepared. Microwave synthesis was used to prepare CdTe QDs coated with mercaptopropionic acid (MPA). MPA ensures biocompatibility and good water solubility of QDs. Therefore QDs can be used directly in bioanalytics. Further, QDs modified with biotinylated glutathione (GSH) were prepared. In this case mercapto group (--SH) ensures a bonding of GSH to QDs surface and biotin serves as high specific linker for streptavidin, avidin or neutravidin modified analyte. Finally, peptide mediated immuno-quantum dots were prepared. CdTe QDs were coated with heptamer HWRGWVC which is able to bind Fc region of human immunoglobulin G.
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