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

提出了一种利用等电聚焦对蛋白质胶体溶液进行连续分馏的新方法。该装置由细胞组成，其细胞由两侧的膜隔开，两性电解质用作缓冲试剂。在不降低分离效率的前提下，通过超滤回收出水中昂贵的两性电解质作为缓冲液再利用。以0.2wt%牛血清白蛋白和0.1wt%牛血红蛋白二元蛋白溶液为过滤样品，研究了细胞内电场强度和流速对分离效率的影响。结果证实，电场强度为12v/cm，保留时间为23mm的条件下，各蛋白的分离效率均在95%以上，两性解液可回收再利用作为缓冲液。在此基础上，从蛋白质的质量平衡出发，建立了预测连续等电聚焦装置分离效率的半理论方程。

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.