Wang Yanli, Chen Quan, Xian Mo, Nian Rui, Xu Fei
College of Life Sciences, Jilin University, Changchun, China.
CAS Key Laboratory of Biobased Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China.
AMB Express. 2018 Jun 1;8(1):93. doi: 10.1186/s13568-018-0622-3.
In recent studies, electronegative multimodal chromatography with Eshmuno HCX was demonstrated to be a highly promising recovery step for direct immunoglobulin G (IgG) capture from undiluted cell culture fluid. In this study, the binding properties of HCX to IgG at different pH/salt combinations were systematically studied, and its purification performance was significantly enhanced by lowering the washing pH and conductivity after high capacity binding of IgG under its optimal conditions. A single polishing step gave an end-product with non-histone host cell protein (nh-HCP) below 1 ppm, DNA less than 1 ppb, which aggregates less than 0.5% and an overall IgG recovery of 86.2%. The whole non-affinity chromatography based two-column-step process supports direct feed loading without buffer adjustment, thus extraordinarily boosting the overall productivity and cost-savings.
在最近的研究中，使用Eshmuno HCX的电负性多模式色谱被证明是从未稀释的细胞培养液中直接捕获免疫球蛋白G（IgG）的一个非常有前景的回收步骤。在本研究中，系统地研究了HCX在不同pH/盐组合下与IgG的结合特性，并且在最佳条件下IgG高容量结合后，通过降低洗涤pH值和电导率，其纯化性能得到显著提高。单一的精制步骤得到的终产品中非组蛋白宿主细胞蛋白（nh-HCP）低于1 ppm，DNA小于1 ppb，聚集体小于0.5%，IgG的总回收率为86.2%。整个基于非亲和色谱的双柱步骤工艺支持直接进料加载而无需缓冲液调整，从而极大地提高了整体生产率并节省了成本。
