Development of a Dynamic Light Scattering Electrobphoresis Apparatus for the Use in the Field of Biochemistry

Research Project

Project/Area Number	60880021
Research Category	Grant-in-Aid for Developmental Scientific Research
Allocation Type	Single-year Grants
Research Field	物質生物化学
Research Institution	Osaka University
Principal Investigator	TAKAGI Toshi Institute for Protein Research, Osaka University, 蛋白質研究所, 教授 (00029943)
Co-Investigator(Kenkyū-buntansha)	KAMEYAMA Keiichi Institute for Protein Research, Osaka University, 蛋白質研究所, 助手 (60177607) 岡宏一大塚電子(株), 部長 SHIRAHAMA Koeishiro Faculty of Cience and Engineering, Saga University, 理工学部, 教授 (50039252) OKA Koichi Ohtsuka Electronics Co. Ltd.
Project Period (FY)	1985 – 1987
Project Status	Completed (Fiscal Year 1987)
Budget Amount *help	¥7,400,000 (Direct Cost: ¥7,400,000) Fiscal Year 1987: ¥400,000 (Direct Cost: ¥400,000) Fiscal Year 1986: ¥2,100,000 (Direct Cost: ¥2,100,000) Fiscal Year 1985: ¥4,900,000 (Direct Cost: ¥4,900,000)
Keywords	Elecltrophoresis / 動的光散乱 / 光散乱 / 蛋白質
Research Abstract	In the field of biochemical research, gel electrophoresis techniques are widely used. Hen this electrophoresis technique is compared with the once widely used Tiselius type electrophoresis, following merits and demerits can be mentioned for the former: (1) merits: simplicity ; high resolution ; minimalization of samples required ; (2) demerits : Low quantitativeness ; non-real time measurement ; damage to samples as the result of detection procedure. Despite of the widely-spread use of electrophoresis, studies on basictopics of electrophoresis and developmennt of new electro- phoresis techniques are not active nowadays. For activation of research in both of the directions, we have planned a development of an instrument, a modern version of Tiselius electro-phoresis apparatus, which can measure electrophoresis of proteins and other biological macromolecules accurately, simply and rapidly. Under support of the present rsearch grant, we have been making effot to realize the plan. The instr … More ument measures the Doppler effect observed with scattered light emitted from an electro- phoresing particle. Such an apparatus is called a dynamic light scattering electrophoresis apparatus. Not a small number of such apparatus have been developed, and a few have been commercialized. All of them were designed to measure electrophoresis of relatively large particles like cells and emulsions. Our apparatus was designed especially for measurement of electrophoresis of proteins and other blomacromolecules which were out of the range of apparatus hitherto developed. We have successfully developed such an apparatus. The apparatus has an optics of heaterodyne type and observes scattering light over an angle of 5 to 30 degrees< Autocorrelation function of the electric field of scattered light is computed by a soft-type photon correlator, and the power spectrum corresponding to an electrophoretic diagram can be obtained by Fourier-transform. An narrow-channel electrophoresis cell was used to be free from convectional disturbance. Measurements were made setting scattering volume at various positions normalto the electric field to reveal the presence of the effect of electroosmosis. Net electrophoretic movement could be measured at the stationary layer for bovine serum albumin and other several proteins. Less