Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China; Key Laboratory of Systems Bioengineering and Frontiers Science Center for Synthetic Biology (Ministry of Education), Tianjin University, Tianjin 300350, China.
Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300350, China; Key Laboratory of Systems Bioengineering and Frontiers Science Center for Synthetic Biology (Ministry of Education), Tianjin University, Tianjin 300350, China.
J Chromatogr A. 2021 Feb 8;1638:461865. doi: 10.1016/j.chroma.2020.461865. Epub 2021 Jan 5.
The strategy of using polymer-grafted media is effective to create protein chromatography of high capacity and uptake rate, giving rise to an excellent performance in high-throughput protein separation due to its high dynamic binding capacity. Taking the scientific development and technological innovation of protein chromatography as the objective, this review is devoted to an overview of polymer-grafted media reported in the last five years, including their fabrication routes, protein adsorption and chromatography, mechanisms behind the adsorption behaviors, limitations of polymer-grafted media and chromatographic operation strategies. Particular emphasis is placed on the elaboration and discussion on the behaviors of ion-exchange chromatography (IEC) with polymer-grafted media because IEC is the most suitable chromatographic mode for this kind of media. Recent advances in both the theoretical and experimental investigations on polymer-grafted media are discussed by focusing on their implications to the rational design of novel chromatographic media and mobile phase conditions for the development of high-performance protein chromatography. It is concluded that polymer-grafted media are suitable for development of IEC and mixed-mode chromatography with charged and low hydrophobic ligands, but not for hydrophobic interaction chromatography with high hydrophobic ligands and affinity chromatography with ligands that have single binding site on the protein.
采用聚合物接枝介质的策略对于创造高容量和高摄取率的蛋白质色谱是有效的，由于其高动态结合能力，在高通量蛋白质分离中表现出优异的性能。本综述以蛋白质色谱的科学发展和技术创新为目标，综述了过去五年中报道的聚合物接枝介质，包括它们的制备路线、蛋白质吸附和色谱、吸附行为背后的机制、聚合物接枝介质的局限性以及色谱操作策略。特别强调了对聚合物接枝介质离子交换色谱（IEC）行为的阐述和讨论，因为 IEC 是最适合这种介质的色谱模式。通过关注对新型色谱介质和用于开发高性能蛋白质色谱的流动相条件的合理设计的意义，讨论了聚合物接枝介质在理论和实验研究方面的最新进展。结论是，聚合物接枝介质适合于开发带有带电和低疏水性配体的 IEC 和混合模式色谱，而不适合于带有高疏水性配体的疏水相互作用色谱和具有蛋白质上单个结合位点的配体的亲和色谱。
