Kundu S, Fenters C
Diagnostics Division, Abbott Laboratories, North Chicago, IL 60064, USA.
J Capillary Electrophor. 1995 Nov-Dec;2(6):273-7.
The isoelectric points (pIs) of murine monoclonal antibodies (MAbs) determined by published one-step capillary isoelectric focusing (CIEF) methods using simultaneous focusing and electroosmotic mobilization showed appreciable deviation from the pI values determined by the slab gel electrophoretic method. The present report describes a modified one-step CIEF method that accurately determines the pIs of MAbs. The modifications include use of internal standards with pIs below and above the pI ranges of MAbs and calculation of pIs from a fitted nonlinear standard curve. Proteins with pIs in the same range as MAbs were first used to establish the efficacy of the method. The present CIEF procedure is highly reproducible and easy to perform for MAbs.
采用同时聚焦和电渗迁移的已发表的一步毛细管等电聚焦（CIEF）方法测定的鼠单克隆抗体（MAb）的等电点（pI），与通过平板凝胶电泳法测定的pI值存在明显偏差。本报告描述了一种改进的一步CIEF方法，该方法能准确测定MAb的pI。改进之处包括使用pI低于和高于MAb pI范围的内标，并根据拟合的非线性标准曲线计算pI。首先使用pI与MAb在同一范围内的蛋白质来确定该方法的有效性。目前的CIEF程序对MAb具有高度可重复性且易于操作。
