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Hyaluronan-albumin system dynamics
Jarábková, Sabína ; Smilek, Jiří (referee) ; Chytil, Martin (advisor)
The bachelor thesis deals with the hyaluronan and albumin system dynamics. In this study was to use these methods: interfacial rheology, tensiometry, method of maximum pressure in the bubble and UV-VIS spectrophotometry. The measurements were used with various concentrations of albumin and a constant concentration of hyaluronan( high molecular and low molecular too) and then conversely. All solutions were measured and the obtained data were compared with the results of BSA in buffer without HA. Solutions were prepared with citrate-phosphate buffer of pH about 4. Were observed interaction between HA and BSA.
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Characterization and stabilization of pancreatin
Wurstová, Agáta ; Němcová, Andrea (referee) ; Obruča, Stanislav (advisor)
This work focuses on a study of enzyme mixture pancreatin, its characterization and subsequent encapsulation into liposomes. As a reference proteins bovine serum albumin and trypsin were used. Characterization of pancreatin consisted of two parts. The first part focuses on optimization of methods for the concentration determination by absorption spectrophotometry using basic methods for identifying proteins (Biuret method, Hartree-Lowry method and Bradford method). Moreover, UV spectrums of the protein were measured. As a method for identification of protein´s molecular weight, SDS-PAGE was used. To identify components of pancreatin, LPLC was employed in two modifications, ion-exchange chromatography and size exclusion chromatography. The second part is dedicated to the characterization of pancreatin as enzyme in terms of pH and temperature optimum for the enzyme activities of protease (pH 9, 8 and 50 °C), amylase (pH 7 and 40 °C) and lipase (pH 7 and 50 °C). The last part of this work aimed at an encapsulation of pancreatin into liposomes and DLS analysis of distribution of particles and their zeta potential. Liposomes did not spontaneously release encapsulated enzyme. To confirm that proteins were successfully entrapped into liposomes, their structure was disrupted by application of phospholipase D. In conclusion, liposomes can be utilized as delivery systems for native enzymes.
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Utilization of interfacial rheology to study of biological systems
Kachlířová, Helena ; Kalina, Michal (referee) ; Smilek, Jiří (advisor)
The aim of this bachalor thesis is to implement and optimization a relatively new method of interfacial rheology. The optimization of this method was realized on two types of interfaces, specifically liquid-liquid using immiscible systems water-chloroform and water-toluene, and water-air. The applicability of this method was tested by using real samples in form of surfactant septonex and biological system represented by protein (bovine serum albumin). The interfacial layers formed by septonex were purely viscous, where the rigidity of the layer increased slightly with concentration until reaching critical micellar concentration and then the rigidity decreased. In comparison with septonex, the interfacial film made of bovine serum albumin showed predominantly elastic behaviour.
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Electrochemical impedance spectroscopy as a nanostructured bioelectrodes characterization method
Vrbová, Eva ; Urbánková, Kateřina (referee) ; Hrdý, Radim (advisor)
Diploma thesis deals of nanostructured surfaces, nanoparticles and electrochemical characterization methods such as cyclic voltammetry, differential pulse voltammetry and electrochemical impedance spectroscopy. The aim of this thesis is a theoretical research issues of production and characterization nanostructured modified electrodes. The practical part is the production of biomodified nanostructured electrodes by anodi- zation W/Al layers with galvanic deposition of gold or deposition of mercury, a modifi- cation of the electrodes by 11-mercaptoundecanoic acid and by bovine serum albumin (BSA). The thesis includes SEM images of nanostructured electrodes contact angle mea- surements of these electrodes and form an electrical circuit with subsequent simulation waveforms.
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Utilization of interfacial rheology to study of biological systems
Kachlířová, Helena ; Kalina, Michal (referee) ; Smilek, Jiří (advisor)
The aim of this bachalor thesis is to implement and optimization a relatively new method of interfacial rheology. The optimization of this method was realized on two types of interfaces, specifically liquid-liquid using immiscible systems water-chloroform and water-toluene, and water-air. The applicability of this method was tested by using real samples in form of surfactant septonex and biological system represented by protein (bovine serum albumin). The interfacial layers formed by septonex were purely viscous, where the rigidity of the layer increased slightly with concentration until reaching critical micellar concentration and then the rigidity decreased. In comparison with septonex, the interfacial film made of bovine serum albumin showed predominantly elastic behaviour.
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Electrochemical impedance spectroscopy as a nanostructured bioelectrodes characterization method
Vrbová, Eva ; Urbánková, Kateřina (referee) ; Hrdý, Radim (advisor)
Diploma thesis deals of nanostructured surfaces, nanoparticles and electrochemical characterization methods such as cyclic voltammetry, differential pulse voltammetry and electrochemical impedance spectroscopy. The aim of this thesis is a theoretical research issues of production and characterization nanostructured modified electrodes. The practical part is the production of biomodified nanostructured electrodes by anodi- zation W/Al layers with galvanic deposition of gold or deposition of mercury, a modifi- cation of the electrodes by 11-mercaptoundecanoic acid and by bovine serum albumin (BSA). The thesis includes SEM images of nanostructured electrodes contact angle mea- surements of these electrodes and form an electrical circuit with subsequent simulation waveforms.
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Hyaluronan-albumin system dynamics
Jarábková, Sabína ; Smilek, Jiří (referee) ; Chytil, Martin (advisor)
The bachelor thesis deals with the hyaluronan and albumin system dynamics. In this study was to use these methods: interfacial rheology, tensiometry, method of maximum pressure in the bubble and UV-VIS spectrophotometry. The measurements were used with various concentrations of albumin and a constant concentration of hyaluronan( high molecular and low molecular too) and then conversely. All solutions were measured and the obtained data were compared with the results of BSA in buffer without HA. Solutions were prepared with citrate-phosphate buffer of pH about 4. Were observed interaction between HA and BSA.
|
|
|
Characterization and stabilization of pancreatin
Wurstová, Agáta ; Němcová, Andrea (referee) ; Obruča, Stanislav (advisor)
This work focuses on a study of enzyme mixture pancreatin, its characterization and subsequent encapsulation into liposomes. As a reference proteins bovine serum albumin and trypsin were used. Characterization of pancreatin consisted of two parts. The first part focuses on optimization of methods for the concentration determination by absorption spectrophotometry using basic methods for identifying proteins (Biuret method, Hartree-Lowry method and Bradford method). Moreover, UV spectrums of the protein were measured. As a method for identification of protein´s molecular weight, SDS-PAGE was used. To identify components of pancreatin, LPLC was employed in two modifications, ion-exchange chromatography and size exclusion chromatography. The second part is dedicated to the characterization of pancreatin as enzyme in terms of pH and temperature optimum for the enzyme activities of protease (pH 9, 8 and 50 °C), amylase (pH 7 and 40 °C) and lipase (pH 7 and 50 °C). The last part of this work aimed at an encapsulation of pancreatin into liposomes and DLS analysis of distribution of particles and their zeta potential. Liposomes did not spontaneously release encapsulated enzyme. To confirm that proteins were successfully entrapped into liposomes, their structure was disrupted by application of phospholipase D. In conclusion, liposomes can be utilized as delivery systems for native enzymes.
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