Separation and Purification of Biochemicals by Affinity Combined with Batch-wise Adsorption and Column-wise Elution

提出了一种亲和分离生化物质的新工艺。吸附阶段由传统的柱式操作改为间歇式操作。在分批模式下，溶质可以从大量非常稀的溶液中快速吸附。然后，分批吸附和柱洗脱的组合过程在短时间内带来了有效的纯化。建立了两种（大、小）尺寸的实验装置。该吸附装置由鼓泡浆池吸附和固定床洗脱组成。气泡用于在吸附器中搅拌，以保护机械强度弱的吸附剂凝胶。溶质在鼓泡浆液罐中被吸附后，吸附剂被转移到下塔以形成固定床。用缓冲溶液洗涤吸附剂，然后用洗脱剂洗脱被吸附的组分。在用缓冲溶液洗涤吸附剂之后，将吸附剂移动到上部罐用于下一吸附阶段。通过使用下部柱进行柱式吸附以与分批式吸附进行比较。采用Blue-SepharoseCL-6 B和大豆胰蛋白酶抑制剂分别纯化白蛋白和胰蛋白酶，并采用离子交换树脂进行纯化。白蛋白和胰蛋白酶的吸附等温线分别为Langmuir型和Freundlich型。从胰蛋白酶的实验结果发现，在间歇式吸附的操作时间是短的0.14倍，在柱式吸附。白蛋白的洗脱浓度与料液浓度之比为14倍，胰蛋白酶的洗脱浓度与料液浓度之比为35倍。因此，从实验和理论结果表明，该组合过程是有效的，以缩短每一个周期的总操作时间。

A new process in affinity separation of biochemicals was proposed. The adsorption stage was subsituted from the conventional column-wise operation to batch-wise operation. In a batch-wise mode, the solute can be adsorbed quickly from a large quantity of very dilute soution. Then, the combined process with batch-wise adsorption and column-wise elution brings a effcient purification in a short time. Two (small and large) sizes of experiemntal apparatus were constructed. It consisted of a bubble slurry tank for adsorption and a fixed bed column for elution. The air bubble is used for stirring in the adsorber to protect mechanically weak adsorbent gel. After the solute is adsorbed in the bubble slurry tank, adsorbent is transferred to the lower column to from a fixed bed. The adsorbent is washed with buffer solution and then the adsorbed component is eluted by the eluent. After the adsorbent is washed with buffer solution, the adsorbent is moved to the upper tank for the next adsorption stage. The column-wise adsorption was carried out by using the lower column to compare with the batch-wise adsorption. Purifications of albumin by Blue-Sepharose CL-6B and trypsin by Soybean Trypsin Inhibitor and desalting by ion-exchange resin were conducted for the combined process. The adsorption isotherms for albumin and trypsin were Langmuir type and Freundlich type, respectively. It was found from experimental results of trypsin that the operating time in the batch-wise adsorption was shorter by 0.14 times than that in the column-wise adsorption. The ratio of the elution coucentration to the feed concentration was 14 times for albumin and it was 35 times for trypsin. Therefore, it was clarified from experimental and theoretical results that the combined process was effective to shorten the total operating time per one cycle.