Kremser Leopold, Bilek Gerhard, Kenndler Ernst
Institute for Analytical Chemistry, University of Vienna, Vienna, Austria.
Electrophoresis. 2007 Oct;28(20):3684-90. doi: 10.1002/elps.200700168.
The different electrophoretic behavior of the members of two groups of proteins with respect to the absence or presence of detergent additives in the BGE was explored. Recombinant soluble concatemers of repeat 3 of the very low density lipoprotein (VLDL)-receptor fused at their N-terminus to maltose-binding protein (MBP) exhibited different electrophoretic mobilities in borate buffer (pH 8.3) in the absence and in the presence of dodecyl-PEG ether (D-PEG). This enabled the separation of the receptor fragments from MBP after enzymatic cleavage. In the presence of SDS, the mobilities of all proteins approached the same values with increase in detergent concentrations. In contrast, viral capsid proteins of a human rhinovirus (HRV) exhibited different migration in the presence of the additive. For the receptor proteins, extreme apparent high plate numbers were observed when the SDS concentration in the sample and the separation buffer differed. This effect might be erroneously interpreted as a high efficiency. However, it is due to the conductivity boundaries caused by the sample and leads to a total loss of separation.
研究了两组蛋白质成员在硼酸盐缓冲液（BGE）中有无去污剂添加剂时的不同电泳行为。在极低密度脂蛋白（VLDL）受体重复序列3的重组可溶性串联体的N端与麦芽糖结合蛋白（MBP）融合，在无十二烷基聚乙二醇醚（D-PEG）和有D-PEG的情况下，其在硼酸盐缓冲液（pH 8.3）中表现出不同的电泳迁移率。这使得酶切后受体片段能与MBP分离。在SDS存在的情况下，随着去污剂浓度增加，所有蛋白质的迁移率接近相同值。相比之下，人鼻病毒（HRV）的病毒衣壳蛋白在添加剂存在时表现出不同的迁移情况。对于受体蛋白，当样品和分离缓冲液中的SDS浓度不同时，会观察到极高的表观塔板数。这种效应可能会被错误地解释为高效率。然而，这是由样品导致的电导率边界造成的，会导致分离完全失败。
