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Functional nanoparticles for plasmonic biosensors
Přítulová, Marie ; Salyk, Ota (referee) ; Vala, Martin (advisor)
This thesis aims to prepare functional gold nanoparticles (AuNPs) and use them in conjunction with a surface plasmon resonance (SPR) biosensor for highly sensitive detection of carcinoembryonic antigen (CEA). In this work, preparation of colloidal AuNPs was investigated and a three-step synthesis was optimized to yield spherical nanoparticles with a diameter of about 100 nm and smooth surface. The synthesized AuNPs were functionalized by a self-assembled monolayer of carboxy-PEG alkanethiols and streptavidin and characterized by UV/VIS spectroscopy and -potential method. Finally, the functionalized AuNPs were employed in sandwich assay for the sensitive detection of CEA and it was demonstrated that they can enhance sensor response to CEA by a factor of 100 compared to the direct detection of CEA.
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In situ study of nuclear acids interactions key for gene expression and therapy based on its silencing
Špringer, Tomáš
In this doctoral thesis we study novel analogues based on R06 aptamers and targeting TAR hairpins of the HIV virus by means of surface plasmon resonance biosensor, which allows for sensitive and real-time monitoring of molecular interactions. We investigate seven different modifications placed at nine different positions on the R06 aptamer in order to find out their applicability in the construction of efficient and stable anti-TAR oligonucleotides. We also determine which positions are suitable for substitutions with a modification and interpret the results in the context of the local nucleotide geometries and interactions in the TAR/anti-TAR complex. In this doctoral thesis we further develop a new fluidic system. This fluidic system eliminates sample dispersion and intermixing effects and thus enables accurate monitoring of molecular interactions on the surface of an SPR chip. We also characterize experimental conditions on the surface of an oligonucleotide chip and their relations towards bio-molecular assays. Specifically, we study the shielding effect of monovalent and divalent cations, which are crucial for the interaction of negatively charged oligonucleotides.
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In situ study of nuclear acids interactions key for gene expression and therapy based on its silencing
Špringer, Tomáš
In this doctoral thesis we study novel analogues based on R06 aptamers and targeting TAR hairpins of the HIV virus by means of surface plasmon resonance biosensor, which allows for sensitive and real-time monitoring of molecular interactions. We investigate seven different modifications placed at nine different positions on the R06 aptamer in order to find out their applicability in the construction of efficient and stable anti-TAR oligonucleotides. We also determine which positions are suitable for substitutions with a modification and interpret the results in the context of the local nucleotide geometries and interactions in the TAR/anti-TAR complex. In this doctoral thesis we further develop a new fluidic system. This fluidic system eliminates sample dispersion and intermixing effects and thus enables accurate monitoring of molecular interactions on the surface of an SPR chip. We also characterize experimental conditions on the surface of an oligonucleotide chip and their relations towards bio-molecular assays. Specifically, we study the shielding effect of monovalent and divalent cations, which are crucial for the interaction of negatively charged oligonucleotides.
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In situ study of nuclear acids interactions key for gene expression and therapy based on its silencing
Špringer, Tomáš ; Štěpánek, Josef (advisor) ; Brynda, Eduard (referee) ; Vorlíčková, Michaela (referee)
In this doctoral thesis we study novel analogues based on R06 aptamers and targeting TAR hairpins of the HIV virus by means of surface plasmon resonance biosensor, which allows for sensitive and real-time monitoring of molecular interactions. We investigate seven different modifications placed at nine different positions on the R06 aptamer in order to find out their applicability in the construction of efficient and stable anti-TAR oligonucleotides. We also determine which positions are suitable for substitutions with a modification and interpret the results in the context of the local nucleotide geometries and interactions in the TAR/anti-TAR complex. In this doctoral thesis we further develop a new fluidic system. This fluidic system eliminates sample dispersion and intermixing effects and thus enables accurate monitoring of molecular interactions on the surface of an SPR chip. We also characterize experimental conditions on the surface of an oligonucleotide chip and their relations towards bio-molecular assays. Specifically, we study the shielding effect of monovalent and divalent cations, which are crucial for the interaction of negatively charged oligonucleotides.
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Functional nanoparticles for plasmonic biosensors
Přítulová, Marie ; Salyk, Ota (referee) ; Vala, Martin (advisor)
This thesis aims to prepare functional gold nanoparticles (AuNPs) and use them in conjunction with a surface plasmon resonance (SPR) biosensor for highly sensitive detection of carcinoembryonic antigen (CEA). In this work, preparation of colloidal AuNPs was investigated and a three-step synthesis was optimized to yield spherical nanoparticles with a diameter of about 100 nm and smooth surface. The synthesized AuNPs were functionalized by a self-assembled monolayer of carboxy-PEG alkanethiols and streptavidin and characterized by UV/VIS spectroscopy and -potential method. Finally, the functionalized AuNPs were employed in sandwich assay for the sensitive detection of CEA and it was demonstrated that they can enhance sensor response to CEA by a factor of 100 compared to the direct detection of CEA.
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