Hou K C, Zaniewski R, Roy S
Life Science Division, Cuno Inc., Meriden, Connecticut 06450.
Biotechnol Appl Biochem. 1991 Apr;13(2):257-68.
Recombinant Protein A was immobilized on a cellulose and acrylic composite matrix through Schiff base formation. Various factors that could affect the binding of immunoglobulin by the Protein A molecules immobilized on the solid matrix were studied to achieve optimum affinity purification. The spacer arm length and ligand concentration of Protein A were verified as factors crucial to optimized IgG purification. Liquid-phase environmental conditions such as pH and salt concentration also play important roles in adsorption capacity by affecting the molecular interaction between IgG and the immobilized Protein A. The rate of interaction between Protein A and IgG is rather fast, with minimal differences observed at 10-fold increases in the cartridge loading rate. This paper describes a cellulose/acrylic composite matrix for immobilizing Protein A, at an optimized ligand concentration, installed on a spacer arm of adequate length, to purify immunoglobulins from animal plasma. The fast-flow property of the cartridge made from such a matrix and its simplicity in operation provide effective means for purifying immunoglobulins on a relatively large scale.
重组蛋白A通过席夫碱形成固定在纤维素和丙烯酸复合基质上。研究了各种可能影响固定在固体基质上的蛋白A分子与免疫球蛋白结合的因素，以实现最佳的亲和纯化。蛋白A的间隔臂长度和配体浓度被证实是优化IgG纯化的关键因素。液相环境条件，如pH值和盐浓度，也通过影响IgG与固定化蛋白A之间的分子相互作用，在吸附能力方面发挥重要作用。蛋白A与IgG之间的相互作用速率相当快，在柱负载率提高10倍时观察到的差异最小。本文描述了一种用于固定蛋白A的纤维素/丙烯酸复合基质，其配体浓度经过优化，安装在长度合适的间隔臂上，用于从动物血浆中纯化免疫球蛋白。由这种基质制成的柱的快速流动特性及其操作简便性为大规模纯化免疫球蛋白提供了有效的手段。
