Spiral Disk Assembly For High-speed Countercurrent Chrom

高速逆流镀铬螺旋盘组件

• 批准号: 6817678
• 负责人: Yoichiro Ito
• 金额: --
• 依托单位: NATIONAL HEART, LUNG, AND BLOOD INSTITUTE
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6817678
• 关键词: biomedical equipment development; peptides; physical separation; protein purification

The spiral disks developed for separating biopolymers by type-J high-speed countercurrent chromatography (HSCCC) showed low resolution in protein samples probably due to a laminar flow effect of the viscous polymer phase system. The spiral disk has an advantage over the conventional multilayer coil separation column in that the channel geometry is easily modified in many ways. It also permits various inserts into the channel to improve its performance. We have found that introducing glass beads or fine Teflon threads into the channel produced intensive carryover of the stationary phase resulting in detrimental loss of the partition efficiency. However, inserting short segments of Teflon tubing into the channel at regular intervals produced positive results. It is assumed that such inserts served as a stream breaker to interrupt the laminar flow of the mobile phase especially for viscous polymer phase systems. A series of experiments was carried out by inserting short segments of Teflon tubing, 3 mm long and 2 mm ID, into the spiral channel of both single and four-spiral disks. The results indicated that in the single spiral disk the partition efficiency was improved by the number of inserts up to 300 which show similar separation with 500 inserts. A four spiral disk with 600 inserts produced the best separation for both dipeptide and protein samples, mainly by improved retention of the stationary phase due to its greater pitch of the sprial. Based on these findings, we have designed "bead-chain spiral disk" (its appearance resembles a bead chain), each consisting of ca 4600 pits (2.7 mm diameter and 2 mm deep) connected with short ducts (0.5 mm long, 2 mm deep and 0.8 to 1.2 mm wide), and examined their performance with a set of test samples by a type-J HSCCC centrifuge. As expected, this new column design substantially improved the separation of both dipeptide and protein samples in terms of stationary phase retention and partition efficiency. Among those, the disk with four spiral channel and wider connection ducts (1.2 mm) produced the best results. Our studies demonstrated the following unique features of the spiral disk when mounted on the type-J CPC: 1. The spiral disk can retain a sufficient amount of polymer phase systems for the separation of biopolymers. 2. The spiral disk with an enhanced pitch permits high flow rates of the mobile phase to shorten the separation times. 3. The performance of the spiral disk can be improved by modifying the channel geometry such as spiral pitch and configuration of the channel. Fabrication of a multiple assembly of the bead-chain sprial disk is currently underway in the NIH Machine shop.

开发用于通过J型高速逆流色谱法（HSCCC）分离生物聚合物的螺旋盘显示出蛋白质样品的分辨率低，这可能是由于粘性聚合物相系统的层流效应所致。螺旋磁盘比常规多层线圈分离柱具有优势，因为通道几何形状很容易通过许多方式修改。它还允许在渠道中进行各种插入以提高其性能。 我们发现，将玻璃珠或细铁氟龙螺纹引入通道中，从而产生了固定相的密集结转，从而导致分区效率的有害损失。然而，将短的特氟龙管的短段定期插入通道会产生积极的结果。假定此类插入物用作断路器，以中断流动相的层流流，尤其是对于粘性聚合物相系统。通过将长3毫米和2毫米ID的Teflon管的短段插入单个和四螺旋盘的螺旋通道，进行了一系列实验。 结果表明，在单个螺旋盘中，通过高达300的插入数量提高了分区效率，这些插入数量显示出500个插入物。一个带有600个插入物的四个螺旋盘可为二肽和蛋白质样品提供最佳的分离，这主要是由于其更大的sprial倾斜，主要是通过改善固定相的保留。 Based on these findings, we have designed "bead-chain spiral disk" (its appearance resembles a bead chain), each consisting of ca 4600 pits (2.7 mm diameter and 2 mm deep) connected with short ducts (0.5 mm long, 2 mm deep and 0.8 to 1.2 mm wide), and examined their performance with a set of test samples by a type-J HSCCC centrifuge.正如预期的那样，根据固定相位保留和分配效率，这种新的柱设计显着改善了二肽和蛋白质样品的分离。其中，具有四个螺旋通道和更宽连接管（1.2毫米）的磁盘可产生最佳效果。 我们的研究表明，在J型CPC上安装时，螺旋磁盘的以下独特特征： 1。螺旋磁盘可以保留足够数量的聚合物相系统来分离生物聚合物。 2。具有增强音高的螺旋磁盘允许流动相的高流速缩短分离时间。 3。可以通过修改通道几何形状（例如螺旋螺距和通道的配置）来提高螺旋盘的性能。 NIH机械车间目前正在进行多个珠链Sprial磁盘的多个组件的制造。