Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Pisa, Italy.
J Chromatogr B Analyt Technol Biomed Life Sci. 2011 Apr 1;879(11-12):756-62. doi: 10.1016/j.jchromb.2011.02.024. Epub 2011 Feb 22.
Matrix metalloproteinase 8 (MMP-8) has been reported to have a key role in several pathologic conditions, like heart diseases, osteoarthritis, multiple sclerosis, and various other inflammatory conditions. Therefore, there is a great interest regarding the development of MMP-8 selective inhibitors. In the recent years, immobilized enzyme reactors (IMERs) proved to be an efficient alternative to solution-based assays. Besides the recycling of the enzyme, IMER approach allows a simple way to determine affinity data and thus the ranking of inhibiting potency of the compounds under study, especially when coupled to MS. In this study, the immobilization of MMP-8 was investigated in terms of type of support, kinetic parameters, storage and pH stability. Epoxy activated silica resulted the best matrix for the preparation of an immobilized enzyme reactor (IMER) containing human MMP-8. The IMER was successfully used for the online screening of known MMP-8 inhibitors in zonal chromatography and inhibition experiments.
基质金属蛋白酶 8（MMP-8）在多种病理情况下发挥着关键作用，如心脏病、骨关节炎、多发性硬化症和各种其他炎症情况。因此，人们对 MMP-8 选择性抑制剂的开发产生了浓厚的兴趣。近年来，固定化酶反应器（IMER）被证明是一种优于基于溶液的测定方法的有效替代方法。除了酶的回收外，IMER 方法还可以简单地确定亲和力数据，从而对研究中化合物的抑制效力进行排序，特别是当与 MS 耦合时。在这项研究中，从支持物的类型、动力学参数、储存和 pH 稳定性等方面研究了 MMP-8 的固定化。环氧基活化硅胶是制备含有人 MMP-8 的固定化酶反应器（IMER）的最佳基质。IMER 成功地用于在线筛选已知的 MMP-8 抑制剂在区域色谱和抑制实验。
