Thömmes J
Institut für Enzymtechnologie der Heinrich-Heine Universität Düsseldorf im Forschungszentrum Jülich, Stetternicher Forst, 52426 Jülich, Germany.
Biotechnol Bioeng. 1999 Feb 5;62(3):358-62. doi: 10.1002/(sici)1097-0290(19990205)62:3<358::aid-bit12>3.0.co;2-9.
A new stationary phase for protein purification was investigated with regard to its performance during capture of selected model proteins. The commercially available matrix consists of a porous agarose backbone, to which dextran is covalently attached. The dextran carries ion-exchange ligands, thus providing a binding space of high ligand density. Breakthrough of various proteins during frontal application to packed beds was measured and the experiments were analyzed in terms of equilibrium and breakthrough capacity. A significant increase of static capacity, as compared with conventional porous matrices, was found. Good dynamic properties allowed utilization of a high percentage of the equilibrium capacity at 10% breakthrough. For all proteins, a decreasing ratio of breakthrough to equilibrium capacity was detected with increasing feed concentration. This observation suggested a significant contribution of solid diffusion to the transport of proteins into the adsorbent particles. The specific architecture of the stationary phase, where the agarose base structure is derivatized with ion-exchange ligand-bearing dextran, may lead to this behavior.
研究了一种用于蛋白质纯化的新型固定相在捕获选定模型蛋白质过程中的性能。市售基质由多孔琼脂糖主链组成，葡聚糖共价连接在其上。葡聚糖带有离子交换配体，从而提供高配体密度的结合空间。测量了各种蛋白质在前沿进样到填充床时的穿透情况，并根据平衡容量和穿透容量对实验进行了分析。与传统多孔基质相比，发现静态容量有显著增加。良好的动态性能使得在10%穿透率时能够利用较高比例的平衡容量。对于所有蛋白质，随着进料浓度的增加，检测到穿透容量与平衡容量的比率降低。这一观察结果表明固体扩散对蛋白质向吸附剂颗粒内的传输有显著贡献。固定相的特定结构，即琼脂糖基础结构用带有离子交换配体的葡聚糖衍生化，可能导致了这种行为。
