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

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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.