Rapid and high-capacity loading of IgG monoclonal antibodies by polymer brush and peptides functionalized microspheres.
Rapid and high-capacity loading of IgG monoclonal antibodies by polymer brush and peptides functionalized microspheres.
复制标题
DOI：
10.1016/j.chroma.2021.461948
复制
发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Dong-xia Hao;Rongyue Zhang;Jia Ge;Peili Ye;Cui Song;Kai Zhu;Xiangming Na;Yongdong Huang;Lan Zhao;Weiqing Zhou;Z. Su;G. Ma
中科院分区：
文献类型：
--
作者：
Dong-xia Hao;Rongyue Zhang;Jia Ge;Peili Ye;Cui Song;Kai Zhu;Xiangming Na;Yongdong Huang;Lan Zhao;Weiqing Zhou;Z. Su;G. Ma
在抗体生产过程中，对现有纯化平台的快速通量和成本降低的要求越来越高。大孔吸附剂在提高大分子物质的纯化通量方面具有广阔的应用前景。与此同时，肽类配体也成为一种很有前途的替代重组蛋白配体的色谱纯化方法。因此，我们设计了一种功能化的微球树脂，其大孔基质为聚合的甲基丙烯酸缩水甘油酯和乙二醇二甲基丙烯酸酯（PGMA-EDMA），配体为六肽（FYEILH）。为了克服肽的空间位阻效应，扩大肽在大孔基质上的结合位点，将肽配体偶联到接枝在微球上的线性PGMA聚合物刷上。与传统的琼脂糖基质树脂和一般的肽接枝微球相比，功能化微球表现出优异的渗透性和快速负载hIgG的高容量，通过在高于110柱体积/小时（CV/h）的快速流速和低于0.5 min的非常短的停留时间下保持稳定的动态结合容量水平。这种功能化的微球提供了一种简便和广泛适用的策略，以开发有吸引力的候选抗体的快速和降低成本的纯化。
Fast-throughput and cost reduction of current purification platforms are becoming increasing requests during antibody manufacture. The macroporous-matrix absorbents have presented extensive potentiality in improving operational throughput during purification of macromolecule. And meanwhile the peptide ligand has become a promising alternative to recombinant protein ligands for cost reduction of chromatographic purification. Therefore, here we designed a functionalized microspheres resin with both macroporous matrix of polymerized glycidyl methacrylate and ethylene glycol dimethacrylate (PGMA-EDMA) and peptide ligand of hexapeptide (FYEILH). In order to circumvent the steric effect of peptides and amplify the binding sites on macroporous matrix, the peptide ligand was coupled on a liner PGMA polymer brushes grafted on microspheres. Comparing to the conventional agarose-matrix resin and the general peptide-grafted microspheres, the functionalized microspheres presented excellent permeability and high capacity to rapid loading hIgG by maintaining a stable level of dynamic binding capacity at fast flow rate above 110 column volume per hour (cv/h) and very short residence time below 0.5 min. Such functionalized microspheres provide a facile and broadly applicable strategy to develop the attractive candidate for rapid and cost-reduced purification of antibody.