Yao Kejian, Yun Junxian, Shen Shaochuan, Wang Lianghua, He Xingjiao, Yu Xiaomei
State Key Laboratory Breeding Base of Green Chemistry Synthesis Technology, College of Chemical Engineering and Materials Science, Zhejiang University of Technology, Hangzhou 310032, China.
J Chromatogr A. 2006 Mar 17;1109(1):103-10. doi: 10.1016/j.chroma.2006.01.014. Epub 2006 Feb 7.
A novel continuous supermacroporous monolithic cryogel embedded with nanometer-size particles was prepared by the radical cryogenic co-polymerization of acrylamide (AAm), N,N'-methylene-bis-acrylamide (MBAAm), allyl glycidyl ether (AGE) and the dispersed surfactant-stabilized Fe3O4 nanoparticles under the freezing-temperature variation condition in a glass column. This special separation matrix has interconnected supermacropores with pore size of 10-50 microm, which permit the free-passage of microbial cells or cell debris in the culture fluids and then is interest in downstream processes. The axial liquid dispersion coefficients of the new continuous supermacroporous monolithic bed at different liquid flow rates were obtained by measuring residence time distributions (RTDs) using tracer pulse-response method. The experimental results showed that the axial liquid dispersion within the bed was weak in a wide water flow rate of 0.5-15 cm/min. The axial dispersion coefficient was found to be increased exponentially with the increase of liquid flow rate. Chromatographic process of bovine serum albumin (BSA) in the cryogel monolithic bed was carried out to reveal the protein breakthrough and elution characteristics. Compared with other reported cryogel beds in literature, the protein adsorption capacity of the present cryogel bed was improved due to the embedded nano-sized solid adsorbents in the gel matrix. Microstructure morphology of the embedded nanoparticles in the cryogel and the gel matrix structure were also analyzed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) in this paper.
通过在玻璃柱中冷冻温度变化条件下，使丙烯酰胺（AAm）、N，N'-亚甲基双丙烯酰胺（MBAAm）、烯丙基缩水甘油醚（AGE）与分散的表面活性剂稳定的Fe3O4纳米颗粒进行自由基低温共聚，制备了一种新型的嵌入纳米尺寸颗粒的连续超大孔整体式冷冻凝胶。这种特殊的分离基质具有相互连通的超大孔，孔径为10 - 50微米，可使微生物细胞或培养液中的细胞碎片自由通过，因此在下游工艺中备受关注。采用示踪剂脉冲响应法测量停留时间分布（RTDs），得到了新型连续超大孔整体式床层在不同液体流速下的轴向液体分散系数。实验结果表明，在0.5 - 15厘米/分钟的宽水流速范围内，床层内的轴向液体分散较弱。发现轴向分散系数随液体流速的增加呈指数增加。对冷冻凝胶整体式床层中牛血清白蛋白（BSA）的色谱过程进行了研究，以揭示蛋白质的穿透和洗脱特性。与文献中报道的其他冷冻凝胶床相比，由于凝胶基质中嵌入了纳米尺寸的固体吸附剂，本冷冻凝胶床的蛋白质吸附能力得到了提高。本文还通过扫描电子显微镜（SEM）和透射电子显微镜（TEM）分析了冷冻凝胶中嵌入纳米颗粒的微观结构形态和凝胶基质结构。
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