接枝多胺聚合物制备大孔阴离子交换色谱介质及其蛋白吸附行为评价  

Preparation of a macroporous anion exchange chromatographic medium by polyamine grafting and evaluation of its protein-adsorption behavior

作　　者：姜泽平 郭旺 李子炀 侯恒扬 霍文迪 王佳一 马磊 靳海波[1] 黄永东[2] 张荣月[1] JIANG Zeping;GUO Wang;LI Ziyang;HOU Hengyang;HUO Wendi;WANG Jiayi;MA Lei;JIN Haibo;HUANG Yongdong;ZHANG Rongyue(College of New Materials and Chemical Engineering,Beijing Institute of Petrochemical Technology,Beijing Key Laboratory of Fuel Cleanliness and Efficient Catalytic Emission Reduction Technology Beijing 102617,China;State Key Laboratory of Biochemical Engineering,Institute of Process Engineering,Chinese Academy of Sciences,Beijing 100190,China)

机构地区：[1]北京石油化工学院新材料与化工学院,燃料清洁化及高效催化减排技术北京市重点实验室,北京102617 [2]中国科学院过程工程研究所生化工程国家重点实验室,北京100190

出　　处：《色谱》2024年第4期360-367,共8页Chinese Journal of Chromatography

基　　金：国家自然科学基金(22074148);国家重点研发计划课题(2021YFC2103401).

摘　　要：以环氧活化聚甲基丙烯酸酯大孔微球(FastSep-epoxy)为基质,通过接枝聚(烯丙基胺)(PAA)制备了大孔阴离子交换色谱介质(FastSep-PAA)。考察了介质的合成条件对离子交换容量(IC)及蛋白结合容量的影响,发现IC随PAA浓度、反应时间和溶液pH的增加均表现为增长趋势;同时结合蛋白吸附容量的变化选择了最优合成条件。通过扫描电子显微镜观察介质的表面形貌,发现其孔隙连通性较好,且接枝前后介质孔道结构无明显变化,PAA配基密度对介质结构无明显影响。此外,通过压汞法和氮气吸附法测定接枝前后不同介质的孔径尺寸和孔径分布情况,并考察该类介质的孔径与蛋白吸附行为的关系,发现其蛋白结合容量未出现随介质孔径尺寸增加而显著下降的现象,且孔径尺寸增加更有利于蛋白在介质内部传质。在126 cm/h的流速下FastSep-PAA介质的原始孔径(即FastSep-epoxy的孔径)为400 nm时的蛋白动态结合容量(DBC)最高(70.3 g/L),该孔径下介质比表面积大,蛋白可吸附位点较多;原始孔径为700 nm及以下的介质蛋白DBC均随流速增加而均有一定下降;原始孔径为1000 nm的介质蛋白DBC几乎不受流速影响,流速由126 cm/h增加至628 cm/h时蛋白DBC仅下降3.5%,且在628 cm/h的流速下能保持57.7 g/L,蛋白分子在该孔径下的扩散速率受限较小,传质性能优异,这一特性表明该类大孔聚合物阴离子交换色谱介质在蛋白高通量分离中具有较大的优势。The macroporous anion exchange chromatographic medium(FastSep-PAA)was prepared through grafting polyallylamine(PAA)onto polyacrylate macroporous microspheres(FastSep-epoxy).The effects of the synthesis conditions,including the PAA concentration,reaction time,and reaction solution pH,on the ion exchange(IC)of the medium were investigated in detail.When the PAA concentration,reaction time,and reaction solution pH were increased,the IC of the medium increased,and optimal synthesis conditions were then selected in combination with changes of protein binding capacity.A scanning electron microscope was used to examine the surface morphology of the medium.The medium possessed high pore connectivity.Furthermore,the pore structure of the medium was preserved after the grafting of PAA onto the macroporous microspheres.This finding demonstrates that the density of the PAA ligands does not appear to have any discernible impact on the structure of the medium;that is,no difference in the structure of the medium is observed before and after the grafting of PAA onto the microspheres.The pore size and pore-size distribution of the medium before and after grafting were determined by mercury intrusion porosimetry and the nitrogen adsorption method to investigate the relationship between pore size(measured in the range of 300-1000 nm)and protein adsorption.When the pore size of the medium was increased,its protein binding capacity did not exhibit any substantial decrease.An increase in pore size may hasten the mass transfer of proteins within the medium.Among the media prepared,that with a pore size of 400 nm exhibited the highest dynamic-binding capacity(DBC:70.3 g/L at 126 cm/h).The large specific surface area of the medium and its increased number of protein adsorption sites appeared to positively influence its DBC.When the flow rate was increased,the protein DBC decreased in media with original pore sizes of less than 700 nm.In the case of the medium with an original pore size of 1000 nm,the protein DBC was independent of the fl

关 键 词：大孔聚合物微球 阴离子交换色谱介质 接枝法 蛋白结合容量 

分 类 号：O658[理学—分析化学]