Harmon B J, Leesong I, Regnier F E
Department of Chemistry, Purdue University, West Lafayette, Indiana 47907-1393.
Anal Chem. 1994 Nov 1;66(21):3797-805. doi: 10.1021/ac00093a041.
Differential electrophoretic mobility between an analyte and its product offers electrophoretically mediated microanalysis (EMMA) a unique capability to selectively control the detection time of the product of a reaction-based chemical analysis. If an analyte and its product differ in migration velocity under the influence of an applied electric field, the apparent velocity and, consequently, the detection time of the product are dependent upon the relative amounts of time the product effectively traverses the capillary with the respective mobilities of the analyte and the product. Consequently, by controlling when the analytical reaction is allowed to occur, the detection time of the reaction product can be selectively maneuvered within a time window defined by the mobilities of the analyte and the product. This paper describes the use of this technique to manipulate the detection time of product profiles independently of nonreacting matrix interferants for the determination of both enzymes and substrates by EMMA. Furthermore, the ability to manipulate product detection times allows for simultaneous EMMA determinations of multiple enzymes or substrates.
分析物与其产物之间的电泳迁移率差异为电泳介导的微分析(EMMA)提供了一种独特的能力,能够选择性地控制基于反应的化学分析产物的检测时间。如果分析物及其产物在施加电场的影响下迁移速度不同,那么产物的表观速度以及检测时间就取决于产物以分析物和产物各自迁移率有效穿过毛细管的相对时间量。因此,通过控制分析反应何时发生,可以在由分析物和产物迁移率定义的时间窗口内选择性地操纵反应产物的检测时间。本文描述了利用该技术独立于非反应性基质干扰物来操纵产物谱的检测时间,以通过EMMA测定酶和底物。此外,操纵产物检测时间的能力允许同时对多种酶或底物进行EMMA测定。