Streptavidin - A Versatile Binding Protein for Solid-Phase Immunoassays

Streptavidin, a tetrameric protein secreted by Streptomyces avidinii, binds tightly to a small growth factor biotin. One of the numerous applications of this high-affinity system comprises the streptavidin-coated surfaces of bioanalytical assays which serve as universal binders for straightforward immobilization of any biotinylated molecule. Proteins can be immobilized with a lower risk of denaturation using streptavidin-biotin technology in contrast to direct passive adsorption. The purpose of this study was to characterize the properties and effects of streptavidin-coated binding surfaces on the performance of solid-phase immunoassays and to investigate the contributions of surface modifications. Various characterization tools and methods established in the study enabled the convenient monitoring and binding capacity determination of streptavidin-coated surfaces. The schematic modeling of the monolayer surface and the quantification of adsorbed streptavidin disclosed the possibilities and the limits of passive adsorption. The defined yield of 250 ng/cm² represented approximately 65 % coverage compared with a modelled complete monolayer, which is consistent with theoretical surface models. Modifications such as polymerization and chemical activation of streptavidin resulted in a close to 10-fold increase in the biotin-binding densities of the surface compared with the regular streptavidin coating. In addition, the stability of the surface against leaching was improved by chemical modification. The increased binding densities and capacities enabled wider high-end dynamic ranges in the solid-phase immunoassays, especially when using the fragments of the capture antibodies instead of intact antibodies for the binding of the antigen. The binding capacity of the streptavidin surface was not, by definition, predictive of the low-end performance of the immunoassays nor the assay sensitivity. Other features such as non-specific binding, variation and leaching turned out to be more relevant. The immunoassays that use a direct surface readout measurement of time-resolved fluorescence from a washed surface are dependent on the density of the labeled antibodies in a defined area on the surface. The binding surface was condensed into a spot by coating streptavidin in liquid droplets into special microtiter wells holding a small circular indentation at the bottom. The condensed binding area enabled a denser packing of the labeled antibodies on the surface. This resulted in a 5 - 6-fold increase in the signal-to-background ratios and an equivalent improvement in the detection limits of the solid-phase immunoassays. This work proved that the properties of the streptavidin-coated surfaces can be modified and that the defined properties of the streptavidin-based immunocapture surfaces contribute to the performance of heterogeneous immunoassays.

Salient Pole Synchronous Machine Modelling in an Industrial Direct Torque Controlled Drive Application

Synchronous motors are used mainly in large drives, for example in ship propulsion systems and in steel factories' rolling mills because of their high efficiency, high overload capacity and good performance in the field weakening range. This, however, requires an extremely good torque control system. A fast torque response and a torque accuracy are basic requirements for such a drive. For large power, high dynamic performance drives the commonly known principle of field oriented vector control has been used solely hitherto, but nowadays it is not the only way to implement such a drive. A new control method Direct Torque Control (DTC) has also emerged. The performance of such a high quality torque control as DTC in dynamically demanding industrial applications is mainly based on the accurate estimate of the various flux linkages' space vectors. Nowadays industrial motor control systems are real time applications with restricted calculation capacity. At the same time the control system requires a simple, fast calculable and reasonably accurate motor model. In this work a method to handle these problems in a Direct Torque Controlled (DTC) salient pole synchronous motor drive is proposed. A motor model which combines the induction law based "voltage model" and motor inductance parameters based "current model" is presented. The voltage model operates as a main model and is calculated at a very fast sampling rate (for example 40 kHz). The stator flux linkage calculated via integration from the stator voltages is corrected using the stator flux linkage computed from the current model. The current model acts as a supervisor that prevents only the motor stator flux linkage from drifting erroneous during longer time intervals. At very low speeds the role of the current model is emphasised but, nevertheless, the voltage model always stays the main model. At higher speeds the function of the current model correction is to act as a stabiliser of the control system. The current model contains a set of inductance parameters which must be known. The validation of the current model in steady state is not self evident. It depends on the accuracy of the saturated value of the inductances. Parameter measurement of the motor model where the supply inverter is used as a measurement signal generator is presented. This so called identification run can be performed prior to delivery or during drive commissioning. A derivation method for the inductance models used for the representation of the saturation effects is proposed. The performance of the electrically excited synchronous motor supplied with the DTC inverter is proven with experimental results. It is shown that it is possible to obtain a good static accuracy of the DTC's torque controller for an electrically excited synchronous motor. The dynamic response is fast and a new operation point is achieved without oscillation. The operation is stable throughout the speed range. The modelling of the magnetising inductance saturation is essential and cross saturation has to be considered as well. The effect of cross saturation is very significant. A DTC inverter can be used as a measuring equipment and the parameters needed for the motor model can be defined by the inverter itself. The main advantage is that the parameters defined are measured in similar magnetic operation conditions and no disagreement between the parameters will exist. The inductance models generated are adequate to meet the requirements of dynamically demanding drives.

Determination of chlorophenols from water by solid phase microextraction - ion mobility spectrometry (SPME-IMS)

Chlorophenols have been classified as possible carcinogens for humans. Chlorophenols have been used as pesticides and wood preservatives. In Finland, during 1930 – 1980s, saw mills used KY-5 wood preservative that contained 2,4,6-TCP, 2,3,4,6-TeCP and PCP. Especially in Finland chlorophenols have entered the environment by leaking from contaminated grounds of old saw mills. Although chlorophenol concentrations found in environment do not cause acute concern, long term exposure can increase the risk of cancer. SPME is relatively cheap and simple sampling method, in which the sample extraction and concentration are performed in a single step. Solvents are not required in SPME. IMS is based on the detection of sample ion drift times. Based on the drift times, reduced mobilities are calculated, which are comparable despite the measurement conditions. SPME-IMS coupling has not been used earlier in the determination of chlorophenols from water samples. The scope of this work was to study, if SPME-IMS system is suitable for detecting chloro-phenols from water samples. The aim was to determine the most optimal extraction condi-tions, which were then applied to real water samples. Following detection limits were deter-mined: 2,4,6-TCP: 0.33 mg/l; 2,3,4,6-TeCP: 0.63 mg/l and PCP: 1.63 mg/l. Detection limits were high compared to the highest possible chlorophenol concentration that is allowed in Finnish drinking water, 10 μg/l. Detected concentrations from water sample differed from verified concentrations in the case of 2,3,4,6-TeCP by 4.6 % and in the case of 2,4,6-TCP by 48.4 %. Based on the results it can be said that SPME-IMS setup is suitable for preliminary analysis of mg/l chlorophenol concentrations from water samples.

Characterization of amine-based CO2 adsorbent for Direct Air Capture

Direct air capture technologies extract CO2 from air at a concentration of as low as 400ppm. The captured CO2 can be used for the production of synthetic methane or liquid fuels. In the literature survey of this thesis, results related to direct air capture by using solid sorbents are presented and critically discussed. In the experimental part, a proprietary amine functionalized resin is characterized for direct air capture. Structural comparison is also made to a commercial resin of similar type. Based on the literature survey, the most important parameters in direct air capture process are low adsorption and desorption temperatures, good cyclic stability in dry and humid conditions, high CO2 outlet purity and a high working capacity. Primary amine functionalized solid sorbents are found to often have good qualities for direct air capture, but overall process performance is rarely studied exhaustively. Based on FTIR spectra, both resin adsorbents are found to be consisted of polystyrene functionalized with primary amine, and capture CO2 by forming carbamate. The commercial resin is more porous, has a slightly higher particle size and contains fewer impurities. Important physical parameters are gained of the proprietary resin, such as internal porosity and median particle size. The resin’s amine group is found to endure thermal treatment reasonably well. CO2 adsorption capacity gained by thermal gravimetry from 400ppm CO2 is highest at 25oC, and is found to be reasonable compared to values presented in literature. Thus, the resin is stated to exhibit promising qualities for direct air capture.

Voiko totuutta tavoittaa? : direct cinema -tyylisuunnan käyttö dokumenttielokuvassa

The subject of this study is the use of direct cinema style in documentary film. The main purpose of this thesis was to the research the ways in which direct cinema style attempts to show and achieve truth in documentary films. The following questions were posed: Is it possible to depict reality in a documentary film; how does the choice of using this style affect the final documentary? The essential purpose of this study was to try to see whether the direct cinema style works when trying to achieve truth in a documentary film. This work consints of two elements, the theoretical part and the short documentary. The theoretical part deals with the history, the truth, and the direct cinema- style in documentaries. The theoretical information of direct cinema has been used when making the short documentary. In the documentary Tuloaula 2 I have studied the way in which using direct cinema -style works in practise. The documentary has followed as strictly as possible the direct cinema style. I was the director, the cameraman and the editor of my documentary film. In the documentary film Tuloaula 2 it appeared that the direct cinema style works best when filming everyday life. By using this style it is easy for the director to observe and leave his own persona in the background. The strength in using the direct cinema style is that it enables the viewer to build his/her own impression on the subject. Even though the direct cinema style aims to achieve objectivity the director has to make numerous subjective choices during both the filming and the editing process. These subjective choices automatically effect the "truth" of the documentary film. The difficulty in a direct cinema style is the large amount of material. This often leads to a long editing phase, which is not often possible in the busy production schedules. The direct cinema style is not at its best when shooting people who are passive because their attention often focuses too much on the camera. In general, the best way to make a documentary film would be to use many documentary styles in one film and not to srictly concentrate on only one style.

Evaluation of the spot urine sampling technique to assess urinary pseudouridine excretion in lactating dairy cows

Selostus: Mahdollisuus lyhytaikaisen virtsankeruun käyttöön lypsylehmien virtsan pseudouridiinin erityksen määrittämisessä

A simple device for sampling of volumetric forest soil cores

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Variation in soil organic carbon and thickness of soil horizons within a boreal forest stand - effect of trees and implications for sampling

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Cut-off importance sampling of bole volume

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