用于肝素与抗凝血酶III动态相互作用研究的涂覆有聚甲基丙烯酸缩水甘油酯的石英晶体微天平流动注射分析

QCM-FIA with PGMA coating for dynamic interaction study of heparin and antithrombin III.

作者信息

Zhang Hongwu, Zhao Rui, Chen Zhiyong, Shangguan Di-Hua, Liu Guoquan

机构信息

Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080, China.

出版信息

Biosens Bioelectron. 2005 Jul 15;21(1):121-7. doi: 10.1016/j.bios.2004.08.041.

DOI:10.1016/j.bios.2004.08.041

PMID:15967359

Abstract

In this work, we describe a method of constructing a film of linear poly(glycidyl methacrylate) (PGMA) polymer onto the surface of quartz crystal microbalance (QCM) electrode as a coating material that allows easy coupling of heparin molecules onto the electrode and facilitates the determination of the interaction between heparin and antithrombin III (AT III). The PGMA film was characterized with atomic force microscopy (AFM) and infra-red spectroscopy. The coupling of heparin was accomplished in one step solution reaction. A home-made quartz crystal microbalance-flow injection analysis (QCM-FIA) system with data analysis software developed in our laboratory was used to determine the interaction. The interactions between immobilized heparin and AT III were studied with various concentrations under various conditions. The obtained constants are kass=(1.49+/-0.12)x10(3)mol-1ls-1, kdiss=(3.94+/-0.63)x10(-2)s-1, KA=(3.82+/-0.33)x10(4)mol-1l.

摘要

在本工作中，我们描述了一种在石英晶体微天平（QCM）电极表面构建线性聚甲基丙烯酸缩水甘油酯（PGMA）聚合物薄膜的方法，该薄膜作为一种涂层材料，能使肝素分子轻松偶联到电极上，并有助于测定肝素与抗凝血酶III（AT III）之间的相互作用。用原子力显微镜（AFM）和红外光谱对PGMA薄膜进行了表征。肝素的偶联通过一步溶液反应完成。使用我们实验室开发的带有数据分析软件的自制石英晶体微天平-流动注射分析（QCM-FIA）系统来测定相互作用。在不同条件下，用不同浓度研究了固定化肝素与AT III之间的相互作用。得到的常数为：结合速率常数kass =(1.49±0.12)×10³mol⁻¹·L⁻¹·s⁻¹，解离速率常数kdiss =(3.94±0.63)×10⁻²s⁻¹，亲和常数KA =(3.82±0.33)×10⁴mol⁻¹·L⁻¹ 。