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[亲和介质和溶液条件对从蛋白质溶液中去除内毒素的影响]

[Effect of affinity medium and solution conditions on endotoxin removal from protein solutions].

作者信息

Xing Haixin, Huang Yongdong, Li Yan, Luo Jian, Zhang Liye, Ma Guanghui, Su Zhiguo

机构信息

College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029, China.

出版信息

Sheng Wu Gong Cheng Xue Bao. 2010 Nov;26(11):1584-95.

Abstract

Endotoxin removal is essential for the safety of biological products. To remove endotoxin efficiently, we used polymyxin B (PMB) affinity adsorbent to remove endotoxin from protein solutions by static adsorption. We studied the effects of spacer length and ligand density of the affinity adsorbent, pH, salt type and concentration, protein type and concentration, endotoxin concentration, and additive on endotoxin removal and protein recovery. Endotoxin content and protein concentration were determined by test and Lowry assay respectively. The results showed that PMB affinity adsorbent had high capacity, high adsorption speed, high removal efficiency and good reusability. In addition, ligand density, pH, salt concentration and the isoelectric point and hydrophobicity of protein all had remarkable influence on the endotoxin removal. Under the optimal conditions, the recoveries of hemoglobin, human serum albumin and lysozyme were 87.2%, 73.4% and 97.3%, respectively, and the corresponding endotoxin removal rates 99.8%, 97.9% and 99.7%, respectively. This study illustrated the effects of solution conditions on the efficiency of endotoxin removal and protein recovery, and would provide useful reference for the efficient removal of endotoxin from biological products.

摘要

内毒素去除对于生物制品的安全性至关重要。为了高效去除内毒素,我们使用多粘菌素B(PMB)亲和吸附剂通过静态吸附从蛋白质溶液中去除内毒素。我们研究了亲和吸附剂的间隔臂长度和配体密度、pH值、盐的类型和浓度、蛋白质的类型和浓度、内毒素浓度以及添加剂对内毒素去除和蛋白质回收率的影响。内毒素含量和蛋白质浓度分别通过检测和Lowry法测定。结果表明,PMB亲和吸附剂具有高容量、高吸附速度、高去除效率和良好的可重复使用性。此外,配体密度、pH值、盐浓度以及蛋白质的等电点和疏水性对内毒素去除均有显著影响。在最佳条件下,血红蛋白、人血清白蛋白和溶菌酶的回收率分别为87.2%、73.4%和97.3%,相应的内毒素去除率分别为99.8%、97.9%和99.7%。本研究阐明了溶液条件对内毒素去除效率和蛋白质回收率的影响,将为从生物制品中高效去除内毒素提供有用的参考。

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