京都大学 / 0048 / 新制・課程博士 / 博士(工学) / 甲第21964号 / 工博第4619号 / 新制||工||1720(附属図書館) / 京都大学大学院工学研究科都市環境工学専攻 / (主査)教授 米田 稔, 教授 高岡 昌輝, 准教授 福谷 哲 / 学位規則第4条第1項該当 / Doctor of Philosophy (Engineering) / Kyoto University / DGAM

<p>Studies of binding reactions are of highest importance in a vast number of areas of biomedicine and biotechnology. A demand for fast and accurate small-volume measurements grows stronger, partly due to the development of therapeutic antibodies. In this report, a novel method for studies of binding reactions of antibodies is described. The use of a microfluidic platform shows promising results in determination of affinity binding constants.</p><p>Affinities between 1E-09 and 1E-11 M have been determined for four TSH antibodies. Reproducibility tests give a CV below 10%, using different Gyrolab instruments and microfluidic CD:s. The method carries the advantages of using solution-based measurements of unmodified molecules. Also an initial proof-of-concept for measurement of binding reaction rate constants shows further usage of the method. The kinetic association rate constant has been determined to 2E+06 M-1s-1 for one antibody. The possibility of using this method for screening of antibody libraries is also discussed.</p>

Studies of binding reactions are of highest importance in a vast number of areas of biomedicine and biotechnology. A demand for fast and accurate small-volume measurements grows stronger, partly due to the development of therapeutic antibodies. In this report, a novel method for studies of binding reactions of antibodies is described. The use of a microfluidic platform shows promising results in determination of affinity binding constants. Affinities between 1E-09 and 1E-11 M have been determined for four TSH antibodies. Reproducibility tests give a CV below 10%, using different Gyrolab instruments and microfluidic CD:s. The method carries the advantages of using solution-based measurements of unmodified molecules. Also an initial proof-of-concept for measurement of binding reaction rate constants shows further usage of the method. The kinetic association rate constant has been determined to 2E+06 M-1s-1 for one antibody. The possibility of using this method for screening of antibody libraries is also discussed.

Due to the increasing demand for processing low-grade ores, larger volumes of material are being processed. Therefore, the size of flotation equipment has significantly increased for the past decades. The studies related to scale-up are and will remain to be crucial in terms of designing larger flotation equipment. One of the most important factors for flotation scaling-up is the “flotation rate constant”. Hence, the main aim of this investigation was to understand the scale-up criteria when the size of different laboratory-scale cells increases, using the Outotec GTK LabCell®. This was done by assessing the influence of impeller speed, as a hydrodynamic variable, on the flotation performance. Recovery was found to increase with an increase in the cell area to rotor diameter ratio. Flotation rate and recovery increased with an increase in the impeller speed until a certain point that it eventually decreased for the 2 l and 7.5 l cells. For the 4 l cell, the flotation rate and recovery decreased with increasing the impeller speed. The impeller speed of 1200 rpm allowed a successful scale-up based on the flotation rate constants and recovery when increasing the size of the cells. Maintaining the impeller speeds constant at 1300 rpm increased the flotation rate constants and recovery when increasing the cell size from both the 2 and 4 l cells to the 7.5 l cell. A further increase in the impeller speed to 1400 rpm also produced the flotation rate constants and recovery to increase as the cell size increased from both the 2 and 4 l cells to the 7.5 l cell. However, when increasing the cell size from 2 l to 4 l, good results were also observed for all impeller speeds. The products concentrate seem to become finer when decreasing the cell size, with only a few exceptions. The recovery of particles larger than 38 μm was found to differ considerably less among the different scales.