Preparation of nanogel-immobilized porous gel beads for affinity separation of proteins: fusion of nano and micro gel materials
Preparation of nanogel-immobilized porous gel beads for affinity separation of proteins: fusion of nano and micro gel materials
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DOI：
10.1038/pj.2014.101
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发表时间：
2015-02-01
期刊：
影响因子：
2.8
通讯作者：
Miura, Yoshiko
中科院分区：
文献类型：
--
作者：
Hoshino, Yu;Arata, Yuka;Miura, Yoshiko
我们描述了纳米凝胶固定的多孔凝胶珠（GB）作为蛋白质纯化介质的应用程序的制备和评价。将与免疫球蛋白G（IgG）的Fc片段结合的纳米凝胶颗粒（NP）通过多点静电相互作用固定在表面含有季铵阳离子的大孔硬GB的孔表面上。使用荧光标记的NP，在Iml GB浆液中不可逆固定的NP的量被确定为类似于30 mg。通过扫描电子显微镜获得的图像确定了NP均匀地固定在孔的表面上而不阻塞大孔。通过NP-IgG相互作用，模型靶蛋白（IgG）被NP-固定的GBs可逆捕获。NP-固定化的GBs作为新型蛋白质亲和纯化介质具有潜在的应用前景，将廉价且稳定的配体与高性能的支持物相结合。
We describe the preparation and evaluation of nanogel-immobilized porous gel beads (GB) for application as a protein purification medium. Nanogel particles (NP) that bind with the Fc fragment of immunoglobulin G (IgG) were immobilized on the pore surface of macroporous hard GB containing quaternary ammonium cations on the surface via multipoint electrostatic interactions. The amount of NPs that were irreversibly immobilized in 1ml of GB slurry was determined to be similar to 30mg using fluorescent-labeled NPs. Images obtained via scanning electron microscopy established that the NPs were uniformly immobilized on the surface of the pores without blocking the macropores. The model target protein (IgG) was reversibly captured by the NP-immobilized GBs through NP-IgG interactions. NP-immobilized GBs have potential applications as novel affinity purification media for proteins, combining inexpensive and stable ligands with high-performance supports.