Development of Continuous Separation and Refinement Equipment for Colloidal Solution of Proteins Using Isoelectric Focusing

Research Project

Project/Area Number	60550658
Research Category	Grant-in-Aid for General Scientific Research (C)
Allocation Type	Single-year Grants
Research Field	化学工学
Research Institution	Gunma University
Principal Investigator	YUKAWA Hisoshi Gunma Univ., Faculty of Eng. (proffesor), 工学部, 教授 (00006967)
Co-Investigator(Kenkyū-buntansha)	HOSHINO Teruhiko Gunma Univ., Faculty of Eng. (asistant), 工学部, 助手 (90008459) KOBAYASHI Kazumasa Gunma Univ., Faculty of Eng. (lecture), 工学部, 講師 (50008512)
Project Period (FY)	1985 – 1986
Project Status	Completed (Fiscal Year 1986)
Budget Amount *help	¥2,300,000 (Direct Cost: ¥2,300,000) Fiscal Year 1986: ¥1,000,000 (Direct Cost: ¥1,000,000) Fiscal Year 1985: ¥1,300,000 (Direct Cost: ¥1,300,000)
Keywords	Isoelectric Focusing / Electrophoresis / Carrier Ampholytes / Proteins / Separation / 精製
Research Abstract	A new continuous fractionation of colloidal solution of proteins is proposed using isoelectric focusing. The device consists of cells whose cell is separated by membranes at the both sides and the ampholytes are used as a buffer reagent. The costly ampholytes are recovered from the effluent by ultrafiltration and reused as buffer without decreasing the separation efficiency. An experimental study has been carried out to investigate the effect of electric field strength and flow rate in the cell on the separation efficiency in the case of using binary proteins solution, 0.2wt% bovine serum albumin and 0.1wt% bovine hemoglobin, as samples of the filtration. It was confirmed from the result that each protein was separated with efficiency above 95% under the condition of 12v/cm of electric field strength and 23mim of retention time, and the ampholytes were able to recover and reuse as buffer. Futhermore a semi-theoretical equation for predicting separation efficiency of the continuous isoelectric focusing apparatus was obtained experimentally based the mass balance of protein.