High yield expression, refolding, and characterization of recombinant interferon α2/α8 hybrids in Escherichia coli
High yield expression, refolding, and characterization of recombinant interferon α2/α8 hybrids in Escherichia coli
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DOI：
10.1016/s1046-5928(03)00187-6
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发表时间：
2003-10-01
影响因子：
1.6
通讯作者：
Foster, GR
中科院分区：
文献类型：
--
作者：
Platis, D;Foster, GR
干扰素（ifn）是一个多效细胞因子家族，用于治疗各种病毒感染和癌症。低成本生产具有高生物学价值的ifn和发现具有改进性质的ifn对于治疗这些疾病以及了解这些化合物的生理功能具有重要意义。我们描述了一种蛋白表达系统，用于在大肠杆菌中以不溶性形式生产ifn α 2、α 8及其杂交体，并结合了一种高效的两步优化的重折叠和组氨酸标签纯化方案。表达的干扰素具有很高的生物学价值，如抗病毒和抗增殖试验所示，其中一些具有比市售干扰素制剂更高的特异性活性，并表现出新的特性。这种高效、优化的蛋白表达方法不仅可以生产单一干扰素亚型，还可以生产几种天然干扰素和杂交干扰素，产量相对较高，成本较低，可用于功能和潜在的临床分析。(C) 2003 Elsevier Science（美国）版权所有。
Interferons (IFNs) are a family of pleiotropic cytokines used for the treatment of various viral infections and cancers. The low-cost production of IFNs with high biological value and the discovery of IFNs with improved properties are important for the treatment of these diseases as well as for understanding the physiological functions of these compounds. We describe a protein expression system for the production of IFNs alpha2, alpha8, and their hybrids in insoluble form in Escherichia coli, coupled to an efficient two-step optimized refolding and histidine-tag purification protocol. The expressed IFNs were of high biological value, as shown in antiviral and antiproliferative assays and some had specific activities higher than those of the commercially available interferon preparations and exhibited novel properties. This time-efficient, optimized protein expression method allows for the production of not just a single interferon subtype but several native and hybrid IFNs with relatively high yield and low cost that can be used in functional and potentially clinical assays. (C) 2003 Elsevier Science (USA). All rights reserved.