A new integrated membrane filtration and chromatographic device
A new integrated membrane filtration and chromatographic device
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DOI：
10.1021/bp049801m
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发表时间：
2005-03-01
影响因子：
2.9
通讯作者：
Luo, RG
中科院分区：
文献类型：
--
作者：
Xu, YK;Sirkar, KK;Luo, RG
为了改善蛋白质分离，已经开发了一种将膜过滤和色谱结合的新型集成装置。该装置基本上由中空纤维过滤模块组成，其壳侧填充有色谱树脂珠。然而，在中空纤维模块出口附近存在基本上不可渗透的涂覆区。该集成装置具有膜过滤和色谱的优点;它还允许直接从发酵液或裂解物中加载色谱介质，并通过循环过程中的后续洗脱步骤分离吸附的蛋白质。进行界面聚合以涂覆中空纤维膜的底部;中空纤维膜的其余部分保持不受影响。肌红蛋白（Mb）和α-乳白蛋白（（X-LA））主要用作二元混合物中的模型蛋白;还研究了Mb和牛血清白蛋白（BSA）的二元混合物。研究了二元蛋白质混合物在超滤膜和微滤膜中的分离行为。实验结果表明，在UF和MF模块中引入不渗透涂层后，穿透时间和蛋白质负载量均显着改善。对于合成酵母发酵液进料，使用具有涂覆区的MF膜在装置中进行四次基于装载-洗涤-洗脱-再平衡的循环运行以分离Mb和a-LA，其间不进行清洁。四次连续运行的Mb和a-LA洗脱曲线几乎可重叠。由于该装置中的较低跨膜通量加上色谱分离期间的周期性洗涤-洗脱，与常规微滤不同，结垢不是问题。
To improve protein separation, a novel integrated device combining membrane filtration and chromatography has been developed. The device basically consists of a hollow fiber filtration module whose shell side is filled with chromatographic resin beads. However, there is an essentially impermeable coated zone near the hollow fiber module outlet. The integrated device enjoys the advantages of both membrane filtration and chromatography; it also allows one to load the chromatographic media directly from the fermentation broth or lysate and separate the adsorbed proteins through the subsequent elution step in a cyclic process. Interfacial polymerization was carried out to coat the bottom section of the hollow fiber membrane; the rest of the hollow fiber membrane remained unaffected. Myoglobin (Mb) and alpha-lactalbumin ((X-LA) were primarily used as model proteins in a binary mixture; binary mixtures of Mb and bovine serum albumin (BSA) were also investigated. Separation behaviors of binary protein mixtures were studied in devices having either an ultrafiltration (UF) or a microfiltration (MF) membrane. Experimental results show that the breakthrough time and the protein loading capacities were dramatically improved after introducing the impermeable coating in both UF and MF modules. For a synthetic yeast fermentation broth feed, four loading-washing-elution-reequilibration-based cyclic runs for separation of Mb and a-LA were performed in the device using a MF membrane with a coated zone without cleaning in between. The Mb and a-LA elution profiles for the four consecutive runs were almost superimposable. Due to lower transmembrane flux in this device plus the periodical washing-elution during the chromatographic separation, fouling was not a problem, unlike in conventional microfiltration.