Katoh Shigeo, Imada Masami, Takeda Naoki, Katsuda Tomohisa, Miyahara Hiroyoshi, Inoue Masaki, Nakamura Shuji
Graduate School of Scinece and Technology, Kobe University, Kobe 657-8501, Japan.
J Chromatogr A. 2007 Aug 17;1161(1-2):36-40. doi: 10.1016/j.chroma.2007.04.023. Epub 2007 Apr 19.
Considering the large molecular size of IgG antibodies widely used for therapeutic applications, the pore size, pore volume and coupling density of silica-based media were optimized for the effective large-scale purification, using an antibody-protein A affinity system. Silica media, with average pore sizes from 70 nm to 140 nm and surface areas of 26-67 m(2)/g, were prepared and coupled with protein A. The static adsorption capacity and dynamic binding capacity of bovine and human IgG were measured at superficial liquid velocities ranging from 94 to 720 cm/h. The volumetric coefficient of mass transfer of the alkali-treated silica-based protein A media, with a pore size of 110 nm, was four times higher than the values for cross-linked agarose media and thus had high dynamic binding capacities at high superficial liquid velocities.
考虑到广泛用于治疗应用的IgG抗体分子尺寸较大，利用抗体-蛋白A亲和系统，对基于二氧化硅的介质的孔径、孔体积和偶联密度进行了优化，以实现有效的大规模纯化。制备了平均孔径为70纳米至140纳米、表面积为26 - 67平方米/克的二氧化硅介质，并与蛋白A偶联。在94至720厘米/小时的表观液体流速下，测量了牛和人IgG的静态吸附容量和动态结合容量。孔径为110纳米的碱处理二氧化硅基蛋白A介质的体积传质系数比交联琼脂糖介质的值高四倍，因此在高表观液体流速下具有高动态结合容量。
