使用自动化多步色谱纯化法生产毫克级的亲和标签蛋白。
Production of milligram quantities of affinity tagged-proteins using automated multistep chromatographic purification.
作者信息
Bhikhabhai Rama, Sjöberg Anna, Hedkvist Lotta, Galin Markus, Liljedahl Pia, Frigård Tuomo, Pettersson Niklas, Nilsson Mats, Sigrell-Simon Jill A, Markeland-Johansson Christine
机构信息
Department of Research and Development, GE Healthcare Company, Amersham Biosciences AB, SE-751 84 Uppsala, Sweden.
出版信息
J Chromatogr A. 2005 Jul 1;1080(1):83-92. doi: 10.1016/j.chroma.2005.05.026.
A new chromatography system, AKTAxpress (GE Healthcare, Amersham Biosciences, Uppsala, Sweden) has been designed to meet the demand for high-throughput purification of proteins in structural genomics and drug discovery. The system offers a number of automated multistep purification protocols for affinity-tagged proteins. All protocols start with affinity chromatography followed by combinations of desalting, ion exchange chromatography and gel filtration. As an option, tag removal can be included in the purification protocols. Up to 16 proteins can be purified per day and the yield can be as high as 50 mg of each protein at > 90% purity. To highlight the versatility of the system, this paper presents several case studies; purifications of hexahistidine- and glutathione S-transferase-tagged proteins using different protocols, automated on-column tag cleavage and optimization studies for a hexahistidine-tagged kinase.
一种新型色谱系统AKTAxpress（通用电气医疗集团，瑞典乌普萨拉阿默沙姆生物科学公司）已被设计用于满足结构基因组学和药物研发中对蛋白质高通量纯化的需求。该系统为带有亲和标签的蛋白质提供了多种自动化多步纯化方案。所有方案均以亲和色谱开始，随后是脱盐、离子交换色谱和凝胶过滤的组合。作为一种选择，标签去除可包含在纯化方案中。每天最多可纯化16种蛋白质，每种蛋白质的产量可达50毫克，纯度>90%。为突出该系统的多功能性，本文展示了几个案例研究；使用不同方案对六聚组氨酸和谷胱甘肽S-转移酶标签蛋白进行纯化、六聚组氨酸标签激酶的自动化柱上标签切割及优化研究。
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