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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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Multifunctional biomolecular assemblies for parallelized analysis of biomolecular interactions
Bocková, Markéta ; Homola, Jiří (advisor) ; Brynda, Eduard (referee) ; Procházka, Marek (referee)
Title: Multifunctional biomolecular assemblies for parallelized analysis of biomolecular interactions Author: Markéta Bocková Department / Institute: Institute of Physics, Charles University Supervisor of the doctoral thesis: Prof. Jiří Homola, Ph.D., DSc., Institute of Photonics and Electronics, The Czech Academy of Sciences Abstract: Surface plasmon resonance (SPR) biosensors represent the most advanced optical method for the direct, real-time monitoring of biomolecular interactions without the need for labelling. This doctoral thesis aims to advance the SPR biosensor method and to expand its utility in the investigation of biomolecular interactions. This encompasses activities on two major fronts of SPR biosensor research - immobilization methods and biosensing methodologies. Methods for the immobilization of biomolecules were researched with the aim of enabling the immobilization of a broad range of biomolecules on the SPR biosensor surface in a spatially controlled manner. The development of novel biosensing methodologies was pursued in order to address the current limitations of SPR biosensors associated with non-specific adsorption and limited analyte transport, and thus to improve the accuracy and robustness of SPR biosensor measurements. Finally, advances in the development of immobilization...
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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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