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磁性琼脂糖载体在液体磁稳定流化床中用于蛋白质吸附的应用。

Application of magnetic agarose support in liquid magnetically stabilized fluidized bed for protein adsorption.

作者信息

Tong Xiao-Dong, Sun Yan

机构信息

Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, Tianjin 300072, China.

出版信息

Biotechnol Prog. 2003 Nov-Dec;19(6):1721-7. doi: 10.1021/bp030028p.

Abstract

A novel magnetic agarose support (MAS) was fabricated for application in a liquid magnetically stabilized fluidized bed (MSFB). It was produced by water-in-oil emulsification method using a mixture of agarose solution and nanometer-sized superparamagnetic Fe(3)O(4) particles as the aqueous phase. The MAS showed good superparamagnetic responsiveness in a magnetic field. A reactive triazine dye, Cibacron blue 3GA (CB), was coupled to the gel to prepare a CB-modified magnetic agarose support (CB-MAS) for protein adsorption. Lysozyme was used as a model protein to test the adsorption equilibrium and kinetic behavior of the CB-MAS. The dependence of bed expansion in the MSFB with a transverse magnetic field on liquid velocity and magnetic field intensity was investigated. Liquid-phase dispersion behavior in the MSFB was examined by measurements of residence time distributions and compared with that obtained in packed and expanded beds. Dynamic lysozyme adsorption in the MSFB was also compared with those in packed and expanded beds. The dynamic binding capacity at 10% breakthrough was estimated at 55.8 mg/mL in the MSFB, higher than that in the expanded bed (31.1 mg/mL) at a liquid velocity of 45 cm/h. The results indicate that the CB-MAS is promising for use in liquid MSFB for protein adsorption.

摘要

制备了一种新型磁性琼脂糖载体(MAS),用于液体磁稳定流化床(MSFB)。它是通过油包水乳化法制备的,以琼脂糖溶液和纳米级超顺磁性Fe(3)O(4)颗粒的混合物作为水相。MAS在磁场中表现出良好的超顺磁性响应。将活性三嗪染料汽巴克隆蓝3GA(CB)偶联到凝胶上,制备用于蛋白质吸附的CB修饰磁性琼脂糖载体(CB-MAS)。以溶菌酶作为模型蛋白,测试CB-MAS的吸附平衡和动力学行为。研究了具有横向磁场的MSFB中床层膨胀对液体流速和磁场强度的依赖性。通过测量停留时间分布来考察MSFB中的液相分散行为,并与填充床和膨胀床中的情况进行比较。还将MSFB中溶菌酶的动态吸附与填充床和膨胀床中的情况进行比较。在液体流速为45 cm/h时,MSFB中10%穿透率下的动态结合容量估计为55.8 mg/mL,高于膨胀床中的(31.1 mg/mL)。结果表明,CB-MAS有望用于液体MSFB中蛋白质的吸附。

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