Takahashi Teruki, Shiyama Takaaki, Hosoya Ken, Tanaka Akito
Chemistry Department, Reverse Proteomics Research Institute Co., Ltd., 2-6-7 Kazusa-Kamatari, Chiba 292-0818, Japan.
Bioorg Med Chem Lett. 2006 Jan 15;16(2):447-50. doi: 10.1016/j.bmcl.2005.09.011. Epub 2005 Nov 14.
Poly(methacrylate) matrices for affinity resins were designed and synthesized based on our previous results that nonspecific protein absorption on affinity resins strongly depended on their hydrophobic property. The novel affinity resins bearing FK506 (6a, 6b) captured specific binding protein, FKBP12, with a small amount of nonspecific binding proteins. The amount of nonspecific binding proteins on 6a-6b was much reduced compared to that on commercially available poly(methacrylate) resins, Toyopearl (8), and was almost the same as that on one of the most popular resins, Affigel (9). Interestingly, 6a and 6b could isolate FKBP52 as a specific binding protein as well, although 8 and 9 could not.
基于我们之前的研究结果,即亲和树脂上的非特异性蛋白质吸附强烈依赖于其疏水性,设计并合成了用于亲和树脂的聚(甲基丙烯酸酯)基质。新型的带有FK506的亲和树脂(6a、6b)能够捕获特异性结合蛋白FKBP12,同时非特异性结合蛋白的量较少。与市售的聚(甲基丙烯酸酯)树脂Toyopearl(8)相比,6a - 6b上的非特异性结合蛋白量大幅减少,并且与最常用的树脂之一Affigel(9)上的量几乎相同。有趣的是,6a和6b也能够分离出作为特异性结合蛋白的FKBP52,而8和9则不能。