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使用前沿色谱法在不同温度下对流化床和填充床柱中的蛋白质吸附进行检测。

Examination of protein adsorption in fluidized bed and packed bed columns at different temperatures using frontal chromatographic methods.

作者信息

Finette G M, Mao Q M, Hearn M T

机构信息

Centre for Bioprocess Technology, Department of Biochemistry and Molecular Biology, Monash University, Wellington Road, Clayton, 3168 Victoria, Australia.

出版信息

Biotechnol Bioeng. 1998 Apr 5;58(1):35-46. doi: 10.1002/(sici)1097-0290(19980405)58:1<35::aid-bit4>3.0.co;2-v.

DOI:10.1002/(sici)1097-0290(19980405)58:1<35::aid-bit4>3.0.co;2-v
PMID:10099259
Abstract

The influences of various experimental parameters on the dynamic adsorption capacity (DAC) and the dynamic adsorption rate (DAR) of a biomimetic affinity silica-based adsorbent in fluidized and packed bed columns operated under plug flow conditions and at different temperatures have been investigated with different inlet concentrations of hen egg white lysozyme (HEWL) and human serum albumin (HSA). The DACs as well as the DARs of both the fluidized and packed beds were examined at 10% saturation (i.e., at the QB value) and the experimental data compared with the corresponding data obtained from batch equilibrium adsorption procedures. Parameters examined included the fluid superficial velocity and protein concentration and their effect on the binding capacity and column efficiency. Consistent with various results reported from this and other laboratories on the behavior of biospecific affinity adsorbents derived from porous silica and zirconia particles, adsorbents prepared from Fractosil 1000 were found to exhibit appropriate rheological characteristics in fluidized bed systems under the experimental conditions. Moreover, changes in temperature resulted in a more significant effect on the breakthrough profiles of HSA compared to HEWL with the immobilized Cibacron Blue F3G-A with Fractosil 1000 adsorbent. This result suggests that temperature effects can possibly be employed profitably in some processes as part of a strategy to enhance column performance with fluidized bed systems for selective recovery of target proteins. At relatively low superficial velocities of the feed, the DARs with HEWL and HSA were similar for both the fluidized and packed bed column systems, whereas, at high superficial velocities, the DARs for these proteins were larger with the packed bed columns.

摘要

研究了在活塞流条件下、不同温度下运行的流化床和填充床柱中,各种实验参数对基于仿生亲和硅胶的吸附剂的动态吸附容量(DAC)和动态吸附速率(DAR)的影响,实验采用了不同入口浓度的鸡蛋清溶菌酶(HEWL)和人血清白蛋白(HSA)。在10%饱和度(即QB值)下检测了流化床和填充床的DAC以及DAR,并将实验数据与分批平衡吸附程序获得的相应数据进行了比较。研究的参数包括流体表观速度和蛋白质浓度及其对结合容量和柱效率的影响。与本实验室和其他实验室报道的关于由多孔二氧化硅和氧化锆颗粒衍生的生物特异性亲和吸附剂行为的各种结果一致,发现在实验条件下,由Fractosil 1000制备的吸附剂在流化床系统中表现出合适的流变特性。此外,与固定化Cibacron Blue F3G-A和Fractosil 1000吸附剂的HEWL相比,温度变化对HSA的穿透曲线影响更大。这一结果表明,在某些过程中,温度效应可能作为一种策略的一部分被有效地利用,以提高流化床系统的柱性能,用于选择性回收目标蛋白质。在进料表观速度相对较低时,流化床和填充床柱系统中HEWL和HSA的DAR相似,而在高表观速度下,填充床柱中这些蛋白质的DAR更大。

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