Capillary Electrophoresis Using Concentrated Solution of Oligomeric Saccharides as the Medium : Elucidation of the Mechanism of the Size-dependent Separation and Its Development

以寡聚糖浓溶液为介质的毛细管电泳：大小依赖性分离机制的阐明及其发展

• 批准号: 07640804
• 负责人: TAKAGI Toshio
• 金额: $ 1.28万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07640804/
• 关键词: Electrophoresis; Protein; Saccharide; Size-dependent separation; キャピラリー; オリゴ糖

The present project originated serendipically from the accidental observation of size-dependent separation of proteins in the presence of SDS of about 0.1% and oligomeric dextran (molecular weight of about 1,000) in a high concentration around 10% by a PhD stident, M.R.Karim and Takagi. The mode of separation was apparently like that in SDS-polyacrylamide. Mobility and viscosity determinations as a function of concentration of the oligomeric dextran clearly showed that the observed size-dependent separation is not due to molecular-sieving, because no sign of the presence of network structure was observed. Under the condition of SDS-polyacrylamide gel electrophoresis, proteins are unfolded to become coiled chains and their polypeptide chains are heavily charged to show free-drainage behavior in free-solution electrophoresis. Such a chains can be naturally size-dependently separated in the presence of molecular-sieve. The new mode of separation we have observed can be interpreted, if the chains loose the free-drainage behavior as the result of the presence of concentrated oligomeric saccharide to become coils not penetrable to solvent and ions contained therein. Confirmation of this working hypothesis is left to be examined, and I will continue making efforts to do so.

本项目的起源是一位博士生 M.R.Karim 偶然观察到，在约 0.1% 的 SDS 和约 10% 的高浓度低聚右旋糖酐（分子量约 1,000）存在下，蛋白质的大小依赖性分离。高木。分离方式显然与SDS-聚丙烯酰胺中的分离方式相似。作为低聚葡聚糖浓度函数的迁移率和粘度测定清楚地表明，观察到的尺寸依赖性分离不是由于分子筛分，因为没有观察到网络结构存在的迹象。在SDS-聚丙烯酰胺凝胶电泳条件下，蛋白质解折叠成卷曲链，其多肽链带大量电荷，在自由溶液电泳中表现出自由排水行为。在分子筛存在下，这种链可以根据尺寸自然地分离。我们观察到的新分离模式是可以解释的，如果由于浓缩低聚糖的存在而导致链失去自由排水行为，变成不可渗透其中所含溶剂和离子的线圈。这个工作假设的确认还有待检验，我将继续努力这样做。

项目成果

Toshiaki Osuga: "Hydrodynamic analysis of electroosmotic flow" J. Phys. Soc. Japan. 65 (#5). 1854-1858 (1996)

Toshiaki Osuga：“电渗流的流体动力学分析”J. Phys。

Toshiaki Osuga: "Hydrodynamic analysis of electroosmotic flow" J.Phys.Soc.Japan. 65(#5). 1854-1858 (1996)

Toshiaki Osuga：“电渗流的流体动力学分析”J.Phys.Soc.Japan。