Centrifugal Precipitation Chromatography

离心沉淀色谱法的发展一直在继续。最近开发的通过将透析管插入到长回旋管中的制备离心沉淀色谱系统具有各种缺点，例如透析膜易碎且容易损坏，并且由于透析管根据柱压的微小变化而改变其体积，因此实验结果不可再现。此外，通过增加管的直径的放大，膜表面与总体积之间的比率急剧减小，导致通过膜的相对传质速率减小。为了提高通过膜的传质效率，我决定使用插入一长段标准塑料管的多段中空纤维过滤器（HFF）管。我发现这种透析设备已经通过两家公司在商业上销售，尽管该设备的长度被限制在几厘米到略超过一米。在第一次试验中，我使用了8个单位的6.5cm长和3mm ID的短HFF管（Spectrum），其与Tygon管互连并安装在铝旋转板的外围。测试运行表明，蛋白质样品倾向于延迟其洗脱，可能是由于沉淀物在连接处的积累。因此，我使用了一个110 cm长得多的HFF（阿默舍姆生物科学公司），它以螺旋方式安装在铝制旋转板上。虽然初步研究的结果显示出一些有希望的结果，但离心机系统遭到破坏。我现在正在等待修理松动的固定伞齿轮和损坏的滚珠轴承。

Development of centrifugal precipitation chromatography has been continued. A preparative centrifugal precipitation chromatographic system recently developed by inserting a dialysis tubing into a long convoluted tubing has various shortcomings such as the dialysis membrance is fragile and easily damaged and the result of the experiment is not reproducible since the dialysis tubing changes its volume according to a subtle change of column pressure. Furthermore, the scaling-up by increasing the diameter of tubing the ratio between the membrane surface and the total volume is sharply decreased, resulting in reduction of a relative mass transfer rate through the membrane. In order to improve the efficincy of the mass transfer through the membrane, I decided to use multiple pieces of hollow fiber filter (HFF) tubing inserted into a long piece of standard plastic tubing. I found that such a dialysis device is already commercially available through two companies, although the length of the unit is limited to several centimeters to slightly over one meter. In the first trial, I used 8 units of short HFF tubes of 6.5cm long and 3 mm ID (Spectrum) which are interconnected with Tygon tubing and mounted at the periphery of an aluminum rotary plate. The test run showed that the protein samples tended to delay in their elution probably due to the accumulation of the precipitates at the junction. Therefore, I used a single piece of much longer HFF of 110 cm (Amersham Biosciences) which is mounted on an aluminum rotary plate in a spiral fashion. Although the result of the pilot studies show some promising results, the centrifuge system was damaged. I am now waiting for the repair of the loosened stationary miter gear and damaged ball bearings.