Khan M Kamran, Luo Jianquan, Khan Rashid, Fan Jinxin, Wan Yinhua
State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China; University of Chinese Academy of Sciences, Beijing 100049, PR China.
J Chromatogr A. 2017 Oct 27;1521:19-26. doi: 10.1016/j.chroma.2017.09.031. Epub 2017 Sep 15.
Fabricating membrane adsorbers with high adsorption capacity and appreciable throughput for the separation and purification of protein products is challenging in biomedical and pharmaceutical industries. Herein, we report the synthesis of a novel membrane adsorber by functionalizing a nylon microfiltration membrane with alginate dialdehyde (ADA) followed by sulphonic addition, without any solvent usage, and its successful application in the purification of lysozyme. Taking advantage of abundant dual cation exchange (CEX) groups on sulphonic-ADA (S-ADA) ligands, this novel S-ADA-nylon membrane adsorber showed an unprecedented static binding capicity of 286mg/mL for lysozyme adsorption. Meanwhile, the prepared membrane adsorber could be easily regenerated (complete protein elution) under mild conditions and be reused at least for five times. Featured with a unique selectivity, the S-ADA-nylon membrane also captured lysozyme from chicken egg white solution with a high purity (100%) and a high recovery of 98%. The purified lysozyme showed similar specific activity as commercial product. The present work provides a facile, green and low-cost approach for the preparation of high-performance membrane adsorbers, which has a great potential in protein production.
在生物医学和制药行业中，制备具有高吸附容量和可观通量以用于蛋白质产品分离和纯化的膜吸附剂具有挑战性。在此，我们报告了一种新型膜吸附剂的合成方法，该方法通过用海藻酸钠二醛（ADA）对尼龙微滤膜进行功能化，随后进行磺酸加成反应，无需使用任何溶剂，并成功将其应用于溶菌酶的纯化。利用磺酸化 - ADA（S - ADA）配体上丰富的双阳离子交换（CEX）基团，这种新型的S - ADA - 尼龙膜吸附剂对溶菌酶吸附表现出前所未有的286mg/mL静态结合容量。同时，制备的膜吸附剂在温和条件下易于再生（蛋白质完全洗脱），并且至少可重复使用五次。S - ADA - 尼龙膜具有独特的选择性，还能从鸡蛋白溶液中捕获溶菌酶，纯度高（100%），回收率达98%。纯化后的溶菌酶显示出与市售产品相似的比活性。本工作为制备高性能膜吸附剂提供了一种简便、绿色且低成本的方法，在蛋白质生产中具有巨大潜力。
