Gu Huimin, Yin Dezhong, Ren Jie, Zhang Baoliang, Zhang Qiuyu
School of Science, Northwestern Polytechnical University, Xi'an 710072, China.
School of Science, Northwestern Polytechnical University, Xi'an 710072, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2016 Oct 15;1033-1034:399-405. doi: 10.1016/j.jchromb.2016.09.020. Epub 2016 Sep 14.
Large size virion is unable to diffuse into pores of conventional porous chromatography particles. Therefore, separation of virion by conventional column-packing materials is not quite efficient. To solve this problem, a monolithic column with large convective pores and quaternary amine groups was prepared and was applied to separate Enterovirus 71 (EV71, ≈5700-6000kDa). Cross-section, pore structure, hydrodynamic performance, adsorption property and dynamic binding capacity of prepared monolithic column were determined. Double-pore structures, macropore at 2472nm and mesopore at 5-60nm, were formed. The porosity was up to 63.3%, which enable higher permeability and lower back pressure of the monolithic column than commercial UNO™ Q1 column. Based on the breakthrough curves, the loading capacity of bovine serum albumin was calculated to be 42.0mg per column. In addition, prepared quaternary amine monolithic column was proved to be suitable for the separation of protein mixture by strong anion-exchange chromatography. As a practical application, prepared monolith column presents excellent performance to the separation of EV71 from virus-proteins mixture.
大尺寸病毒粒子无法扩散到传统多孔色谱颗粒的孔隙中。因此，用传统的柱填充材料分离病毒粒子效率不高。为了解决这个问题，制备了一种具有大对流孔隙和季胺基团的整体柱，并将其应用于分离肠道病毒71型（EV71，约5700 - 6000kDa）。测定了制备的整体柱的横截面、孔结构、流体动力学性能、吸附性能和动态结合容量。形成了双孔结构，大孔为2472nm，中孔为5 - 60nm。孔隙率高达63.3%，这使得整体柱比商业UNO™ Q1柱具有更高的渗透率和更低的背压。根据穿透曲线，计算出每根柱牛血清白蛋白的负载量为42.0mg。此外，制备的季胺整体柱被证明适用于通过强阴离子交换色谱分离蛋白质混合物。作为实际应用，制备的整体柱在从病毒 - 蛋白质混合物中分离EV71方面表现出优异的性能。
