Key Laboratory of Biomass Chemical Engineering of Ministry of Education, College of Chemical and Biological Engineering, Zhejiang University, Hangzhou, P. R. China.
J Sep Sci. 2020 Oct;43(19):3816-3823. doi: 10.1002/jssc.202000325. Epub 2020 Sep 3.
Short peptide biomimetic affinity chromatography as a novel antibody separation chromatography is a potential alternative to protein A chromatography. However, if directly attaching ligand to matrix, the adsorption capacity and mass transfer rate would be affected by pore blockage and steric effect. Grafting resin is an effective method to solve this problem by using polymer as a bridge between matrix and ligand. In this work, a novel resin was prepared by grafting a tetrapeptide to the dextran-grafted matrix. Then, the adsorption properties for human immunoglobin G and BSA were determined. The results showed the saturation adsorption capacity could reach to 133 mg/g resin at pH 8.9 with a significantly low dissociation constant (0.03 mg/mL). The influence of flow rates to dynamic binding capacity of this resin was less than that of the non-grafted resin. The separation performance of the resin showed monoclonal antibody could be well isolated from the Chinese hamster ovary culture supernatant at pH 9.0 with the purity of 93.0% and yield of 84.7% by one step. Overall, this resin could achieve higher binding capacity by the possible of gaining higher ligand density, indicating its potential significance for separation in larger scale systems.
短肽仿生亲和层析作为一种新型的抗体分离层析，是蛋白 A 层析的潜在替代品。然而，如果将配体直接连接到基质上，吸附容量和传质速率会受到孔阻塞和空间位阻的影响。接枝树脂是一种有效的方法，可以通过聚合物作为基质和配体之间的桥梁来解决这个问题。在这项工作中，通过将四肽接枝到葡聚糖接枝的基质上，制备了一种新型的树脂。然后，测定了其对人免疫球蛋白 G 和 BSA 的吸附性能。结果表明，在 pH 8.9 时，其饱和吸附容量可达 133mg/g 树脂，解离常数（0.03mg/mL）显著降低。该树脂的动态结合容量受流速的影响小于非接枝树脂。该树脂在 pH 9.0 时，一步法可从中国仓鼠卵巢培养液中分离出纯度为 93.0%、收率为 84.7%的单克隆抗体，表明其具有良好的分离性能。总之，该树脂通过获得更高的配体密度，可能实现更高的结合容量，表明其在更大规模系统中的分离具有潜在意义。
