Acevedo F
Department of Dermatology, Karolinska Institute, Stockholm.
Electrophoresis. 1993 Oct;14(10):1019-22. doi: 10.1002/elps.11501401162.
The separation of proteins from human serum by isotachophoresis in sucrose density gradients, with mixtures of discrete amphoteric substances as spacers, is described. Open columns and columns with a dialysis membrane to hold the sucrose gradients were used. A simple algorithm based on the Kohlrausch function was used to calculate the amount of each spacer. The pH gradients generated in open columns were found to be in agreement with the calculations. The load was up to two gram proteins. The analysis of the fractions obtained after the separation showed a distribution of components similar to as analytical isotachophoresis. It is concluded that sucrose density gradients are suitable as supporting media for the preparative separation of proteins by isotachophoresis. The high resolution attained and the possibility of scaling-up the separation systems are major advantages of this system. In addition, the sample is easily and completely recoverable.
本文描述了在蔗糖密度梯度中，以离散两性物质混合物作为间隔离子，通过等速电泳从人血清中分离蛋白质的方法。使用了开放式柱和带有透析膜以保持蔗糖梯度的柱。基于科尔劳施函数的简单算法用于计算每种间隔离子的量。发现开放式柱中产生的pH梯度与计算结果一致。上样量高达两克蛋白质。对分离后得到的馏分进行分析，结果显示其成分分布与分析性等速电泳相似。得出结论，蔗糖密度梯度适合作为通过等速电泳进行蛋白质制备分离的支持介质。该系统的主要优点是具有高分辨率以及放大分离系统的可能性。此外，样品易于且能完全回收。
