Use of mobility ratios to estimate binding constants of ligands to proteins in affinity capillary electrophoresis
Use of mobility ratios to estimate binding constants of ligands to proteins in affinity capillary electrophoresis
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DOI：
10.1016/s0378-4347(98)00161-3
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发表时间：
1998-09-11
期刊：
影响因子：
--
通讯作者：
Gomez, FA
中科院分区：
文献类型：
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作者：
Kawaoka, J;Gomez, FA
本工作评估了使用迁移率（M）估计蛋白质的结合常数的配体使用亲和毛细管电泳（ACE）。使用两种模型系统证明了这一概念：来自东方链霉菌的万古霉素（货车）和碳酸酐酶B（CA B，EC 4.2.1.1）。在广泛的情况下，M（Δ M）随配体浓度[L]的变化对Δ M的曲线图比传统的迁移率Δ μ随[L]的变化对Δ μ的曲线图更有用地表示毛细管电泳（CE）中的Scatchard曲线，特别是当比较在电渗流存在实质性变化的情况下获得的电泳图时。改变CE系统的电压和/或毛细管长度仅产生M的微小变化，但μ形式分析所使用的更标准的迁移测量的变化要大得多。M在Scatchard分析中的使用提供了一种新的方法来使用ACE估计配体与蛋白质的结合常数。(C)1998 Elsevier Science B. V.保留所有权利。
This work evaluates the use of mobility ratios (M) to estimate binding constants of proteins to ligands using affinity capillary electrophoresis (ACE). This concept is demonstrated using two model systems: vancomycin (Van) from Streptomyces orientalis and carbonic anhydrase B (CAB, EC 4.2.1.1). A plot of change in M (Delta M) over the concentration of ligand [L] versus Delta M yields a more useful representation of the Scatchard plot in capillary electrophoresis (CE) than traditional plots of the change in mobility Delta mu over [L] versus Delta mu in a wide set of circumstances, especially when comparing electropherograms obtained in the presence of substantial variations in electroosmotic flow. Altering the voltage and/or capillary length of the CE system produced only small variations in M, but much larger changes in the more standard measures of migration used by the mu form of analysis. The use of M in the Scatchard analysis offers a new approach to estimating binding constants of ligands to proteins using ACE. (C) 1998 Elsevier Science B.V. All rights reserved.