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

提出了一种新的生物化学亲和分离工艺。吸附阶段由传统的塔式操作改为批量操作。在批量模式下，溶质可以从大量非常稀的溶液中快速吸附。然后，采用间歇式吸附和柱式洗脱相结合的工艺，在短时间内实现了高效的净化。搭建了小、大两种尺寸的实验装置。它由一个用于吸附的泡浆槽和一个用于洗脱的固定床柱组成。利用气泡在吸附器内进行搅拌，保护机械弱的吸附凝胶。溶质在泡浆槽内吸附后，吸附剂从固定床上转移到下塔。用缓冲溶液洗涤吸附剂，然后用洗脱液洗脱被吸附的组分。在用缓冲溶液洗涤吸附剂后，吸附剂被移动到上槽进行下一个吸附阶段。采用下柱进行柱式吸附，并与批量吸附进行比较。用Blue-Sepharose CL-6B纯化白蛋白，用大豆胰蛋白酶抑制剂纯化胰蛋白酶，并用离子交换树脂脱盐。白蛋白和胰蛋白酶的吸附等温线分别为Langmuir型和Freundlich型。胰蛋白酶的实验结果表明，间歇式吸附比柱式吸附的操作时间缩短了0.14倍。洗脱浓度与饲料浓度之比，白蛋白为14倍，胰蛋白酶为35倍。因此，实验和理论结果表明，该组合工艺可以有效缩短每周期总运行时间。

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.