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Study of biomolecular interactions by the method of surface plasmon resonance
Bocková, Markéta ; Homola, Jiří (advisor)
Title: Study of biomolecular interactions by the method of surface plasmon resonance Author: Markéta Bocková Department: Department of Biochemistry, Faculty of Science Supervisor: Prof. Jiří Homola, Ph.D., DSc., Institute of Photonics and Electronics of the Czech Academy of Sciences Abstract: In this diploma work, a surface plasmon resonance (SPR) biosensor technology was exploited to study two different classes of molecular interactions: hybridization of partially complementary oligonucleotides (ONs) and interactions between enzymes and peptides involved in pathogenesis of Alzheimer disease (AD). In particular, the study of interactions between ONs was focused on determining kinetic parameters of the hybridization proces. It was found that the association equilibrium constant decreases with the increasing surface probe density. An SPR biosensor-based method for rapid and sensitive detection of the 17-HSD10 enzyme and its complex with amyloid β (Aβ) peptides was designed and developed. It was demonstrated that the developed method is capable of detecting 17β-HSD10/Aβs complex in real cerebrospinal fluid (CSF) samples and that CSF samples from AD patients exhibited elavated levels of 17β- HSD10/Aβs complex compared to the control groups. The research reported in this thesis resulted in two publications in...
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Study of Biomolecular Interactions with Surface Plasmon Resonance Biosensors
Šípová, Hana ; Homola, Jiří (advisor) ; Houska, Milan (referee) ; Skládal, Petr (referee)
Surface plasmon resonance (SPR) biosensors represent one of the most advanced sensing technologies for real-time studies of biomolecular interactions. In this thesis, methods for functionalization of SPR substrates were optimized and studied via spectroscopic methods. Effects related to the SPR sensor microfluidic interface on the measured biomolecular interactions were analyzed, and furthermore, means to decrease mass-transport limitations were proposed. Several SPR-based assays regarding the detection of nucleic acids were developed, which allow for the detection of physiologically relevant concentrations of nucleic acids as well as point mutations in a nucleic acid sequence. Assays for the determination of the enzymatic activity of HIV integrase and ribonuclease H were developed. These assays can be employed for the design and synthesis of molecules that function either as antiviral drugs or as gene-regulating agents.
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Complex diffractive structures for surface plasmon resonance sensors
Vala, Milan ; Homola, Jiří (advisor) ; Richter, Ivan (referee) ; Procházka, Marek (referee)
Title: Complex diffractive structures for surface plasmon resonance sensors Author: Milan Vala Institute of Photonics and Electronics ASCR, v.v.i. Supervisor of the doctoral thesis: Prof. Jiří Homola, Ph.D., DSc. Institute of Photonics and Electronics, ASCR, v.v.i. Abstract: Nowadays, biosensors based on surface plasmon resonance (SPR) present one of the most advanced label-free optical biosensor technologies. These sensors are based on the monitoring of refractive index changes in the vicinity of a plasmonic metal and enable both rapid and sensitive detection of various biological or chemical agents as well as the analysis of biomolecular interactions in real time. This doctoral thesis presents the results of research of advanced diffractive structures having benefits for SPR biosensor technology. In addition to a theoretical analysis, this work consists of the fabrication and characterization of selected diffractive structures, and furthermore, the development of novel types of SPR biosensors based on these structures. In particular, the results presented in this thesis include the development of a novel method of fabrication of periodic plasmonic arrays based on multiple-beam interference lithography, the development of compact high-resolution diffraction grating-based SPR devices, and development of...
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Surface plasmons in optical microstructures and their sensor applications
Adam, Pavel ; Homola, Jiří (advisor) ; Brynda, Eduard (referee) ; Richter, Ivan (referee)
Title: Surface plasmons in optical microstructures and their sensor applications Author: Pavel Adam Institute: Institute of Photonics and Electronics AS CR, v.v.i., Department of Optical Sensors Supervisor of the doctoral thesis: doc. Ing. Jiří Homola, CSc., DSc. Abstract: This work is focused on the study of surface plasmon resonance (SPR) sensor platforms based on wavelength division multiplexing (WDM) of multiple surface plasmons (SPs). These sensors are based on advanced diffraction gratings supporting either conventional or Bragg-scattered SPs, which are simultaneously excited at different wavelengths. These SPs are studied both analytically and numerically using rigorous coupled-wave analysis and an integral approach. WDM of two and three SPs is presented and followed by the method for the analysis of the resolution, noise and cross-sensitivity. This method is employed to analyze the ability of different SPR sensor platforms (supporting WDM of two SPs) to discriminate refractive index (RI) changes in a thin layer at the sensor surface from background RI changes. The WDM SPR sensors based on advanced diffraction gratings prepared by interferometric holography are developed and tested in a model biosensing experiment consisting of the layer-by-layer growth of protein multilayers. The linear WDM of two...
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Rich Information Optical Biosensor Based on Surface Plasmon Resonance
Dostálek, Jakub ; Homola, Jiří (advisor) ; Richter, Ivan (referee) ; Vrbová, Miroslava (referee)
In the presented thesis, new optical biosensor platform for high-throughput label-free monitoring of biomolecular interactions is reported. This biosensor platform is based on a spectroscopy of surface plasmons on an array of miniature diffraction gratings. Each miniature grating serves as an independent sensing channel and allows for monitoring of optical density changes induced by biomolecular interactions on the grating surface. The presented platform has a potential for monitoring of hundreds of interactions on a single sensor chip. The presented research include theoretical study of surface plasmon resonance on metallic diffraction gratings and the analysis of diffraction grating-based SPR biosensor. In the experimental work, preparation, replication and characterization of diffraction gratings is carried out. A prototype SPR sensor device including an SPR sensor chip with a twodimensional array of sensing channels, fluidic system and SPR chip reader is developed and its main performance characteristics are determined.
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Vedené vlny v periodických strukturách a jejich senzorové aplikace
Vala, Milan ; Homola, Jiří (advisor) ; Richter, Ivan (referee)
This work is introduced by the description of fundamental properties of surface plasmons, propagating along the metal-dielectric interface and along the structure with thin metal film surrounded by the dielectrics with identical refractive indices. It is shown, that the excitation of two coupled modes (long-range surface plasmon - LRSP and short-range surface plasmon - SRSP) on symmetrical structure with a thin metal film is possible. Further, a brief overview of surface plasmons applications in optical sensors is presented. The diffractive multi-layer structure supporting LRSP and SRSP modes, which consists of the thin gold film surrounded by the dielectrics with slightly different refractive indices is designed. The diffractive structure is optimized using rigorous integral method for an application in SPR sensor. Diffractive structures with optimal parameters are prepared and characterized optically and using atomic force microscopy. In the end, an application of developed diffractive structure in SPR sensor with wavelength modulation is demonstrated.
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