Karlsson Robert, Katsamba Phinikoula S, Nordin Helena, Pol Ewa, Myszka David G
Biacore AB, Uppsala, Sweden.
Anal Biochem. 2006 Feb 1;349(1):136-47. doi: 10.1016/j.ab.2005.09.034. Epub 2005 Oct 13.
The classical method of measuring binding constants with affinity-based biosensors involves testing several analyte concentrations over the same ligand surface and regenerating the surface between binding cycles. Here we describe an alternative approach to collecting kinetic binding data, which we call "kinetic titration." This method involves sequentially injecting an analyte concentration series without any regeneration steps. Through a combination of simulation and experimentation, we show that this method can be as robust as the classical method of analysis. In addition, kinetic titrations can be more efficient than the conventional data collection method and allow us to fully characterize analyte binding to ligand surfaces that are difficult to regenerate.
使用基于亲和力的生物传感器测量结合常数的经典方法包括在同一配体表面测试几种分析物浓度，并在结合循环之间再生表面。在此，我们描述了一种收集动力学结合数据的替代方法，我们称之为“动力学滴定”。该方法包括依次注入分析物浓度系列，无需任何再生步骤。通过模拟和实验相结合，我们表明该方法与经典分析方法一样可靠。此外，动力学滴定可能比传统数据收集方法更有效，并且使我们能够全面表征分析物与难以再生的配体表面的结合情况。
