Production of milligram quantities of affinity tagged-proteins using automated multistep chromatographic purification

被引：30

作者：

Bhikhabhai, R ^{[1
]}

Sjöberg, A ^{[1
]}

Hedkvist, L ^{[1
]}

Galin, M ^{[1
]}

Lijedahl, P ^{[1
]}

Frigård, T ^{[1
]}

Pettersson, N ^{[1
]}

Nilsson, M ^{[1
]}

Sigrell-Simon, JA ^{[1
]}

Markeland-Johansson, C ^{[1
]}

机构：

[1] GE Healthcare Co, Dept Res & Dev, Amersham Biosci AB, SE-75184 Uppsala, Sweden

来源：

JOURNAL OF CHROMATOGRAPHY A | 2005年 / 1080卷 / 01期

关键词：

AKTAxpress; hexahistidine; GST; affinity-tagged proteins; automated multistep chromatography; on-column cleavage; protein purification;

D O I：

10.1016/j.chroma.2005.05.026

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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. (c) 2005 Elsevier B.V. All rights reserved.

引用

页码：83 / 92

页数：10

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