Rapid preparation of 1-vinylimidazole based non-affinity polymers for the highly-selective purification of antibodies from multiple biological sources
Rapid preparation of 1-vinylimidazole based non-affinity polymers for the highly-selective purification of antibodies from multiple biological sources
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快速制备 1-乙烯基咪唑基非亲和聚合物，用于高度选择性纯化多种生物来源的抗体
DOI：
10.1016/j.chroma.2020.461607
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发表时间：
2020
影响因子：
4.1
通讯作者：
Jiang Zhengjin
中科院分区：
文献类型：
--
作者：
Lei yutian;Liu xiao;Lu li;Liu cuihua;Xu rongrong;Huang shengfeng;Shen yuan;Deng chunping;Yu jinchen;Zhang tingtiing;Crommen Jacques;Wang qiqin;Jiang Zhengjin
迫切需要开发成本低廉、产能满意的先进纯化技术，以应对当前单克隆抗体（mAb）下游纯化的挑战。在此，首次提出一种简单且廉价的氮杂环分子1-乙烯基咪唑（VIM）作为抗体的捕获配体。然后通过一步聚合制备相应的基于 VIM 的非亲和性聚合物材料 (polyVIM)，用于抗体的高度选择性纯化。与之前报道的材料相比，这种新型材料在不明显牺牲选择性的情况下表现出许多优点，例如更简单、更快速的制造（1.5小时内）、相当甚至更高的结合容量（饱和静态吸附容量> 190 mg/g聚合物，动态结合容量约31.62 mg/g聚合物）、更低的非特异性蛋白质吸附和更低的成本。值得注意的是，polyVIM可以有效地纯化多种生物来源的抗体，纯度高（细胞培养基中的mAb为95.4％，人血清中的hIgG为93.3％），回收率可接受（mAb为91.6％，hIgG为77.0％），并且可重复使用性良好（> 10次）。此外，目标ELISA结合测定和NFAT-luc报告基因测定表明，富集的抗体在整个纯化过程中能够很好地保持其结合活性和生物活性。 PolyVIM材料的优异性能可能归因于VIM对抗体的高识别能力，以及多孔聚合物的生物相容性和防污性能。这项研究为下游工艺中 mAb 的纯化和人血清中 hIgG 的富集提供了一种有前途的替代材料。
There is an urgent need for developing advanced purification techniques with the merits of low cost and satisfactory capacity in order to meet the challenges in the current downstream purification of monoclonal antibodies (mAbs). Herein, a simple and inexpensive nitrogen heterocycle molecule, 1-vinylimidazole (VIM), was proposed as the capture ligand of antibodies for the first time. The corresponding VIM-based non-affinity polymeric material (polyVIM) was then fabricated via a one-step polymerization for use in the highly selective purification of antibodies. Compared to the previously reported materials, this novel material exhibited many advantages without clearly sacrificing selectivity, such as a simpler and faster fabrication (within 1.5 h), comparable or even higher binding capacity (saturated static adsorption capacity > 190 mg/g polymer, dynamic binding capacity about 31.62 mg/g polymer), lower non-specific protein adsorption, and much lower cost. Notably, the polyVIM can effectively purify the antibodies from multiple biological sources with high purity (95.4% for mAbs in the cell culture medium, 93.3% for hIgG in the human serum), with an acceptable recovery (91.6% for mAbs, 77.0% for hIgG), and good reusability (> 10 times). Moreover, the target ELISA binding assay and NFAT-luc reporter gene assay demonstrated that the enriched antibodies can well maintain their binding activity and bioactivity during the whole purification process. The excellent performance of the polyVIM material may be attributed to the high recognition ability of VIM for antibodies, as well as the biocompatible and antifouling properties of the porous polymer. This study provides a promising alternative material for the purification of mAbs in downstream processes and the enrichment of hIgG in human serum.