A novel gigaporous GSH affinity medium for high-speed affinity chromatography of GST-tagged proteins
文献类型：期刊论文
| 作者 | Huang, Yongdong; Zhang, Rongyue; Li, Juan; Li, Qiang; Su, Zhiguo; Ma, Guanghui |
| 刊名 | PROTEIN EXPRESSION AND PURIFICATION
 |
| 出版日期 | 2014-03-01 |
| 卷号 | 95期号:1页码:84-91 |
| 关键词 | GSH affinity chromatography Gigaporous polystyrene microspheres High-speed chromatography GST-tagged proteins |
| ISSN号 | 1046-5928 |
| 其他题名 | Protein Expr. Purif. |
| 中文摘要 | Novel GSH-AP (phenoxyl agarose coated gigaporous polystyrene, Agap-co-PSt) microspheres were successfully prepared by introducing GSH ligand into hydrophilic AP microspheres pre-activated with 1,4-butanediol diglycidyl ether. The gigaporous structure and chromatographic properties of GSH-AP medium were evaluated and compared with commercial GSH Sepharose FF (GSH-FF) medium. The macropores (100-500 nm) of gigaporous PSt microspheres were well maintained after coating with agarose and functionalized with GSH ligand. Hydrodynamic experiments showed that GSH-AP column had less backpressure and plate height than those of GSH-FF column at high flow velocity, which was beneficial for its use in high-speed chromatography. The presence of flow-through pores in GSH-AP microspheres also accelerated the mass transfer rate of biomolecules induced by convective flow, leading to high protein resolution and high dynamic binding capacity (DEC) of glutathione S-transferase (GST) at high flow velocity. High purity of GST and GST-tagged recombinant human interleukin-1 receptor antagonist (rhIL-1RA) were obtained from crude extract with an acceptable recovery yield within 1.5 min at a velocity up to 1400 cm/h. GSH-AP medium is promising for high-speed affinity chromatography for the purification of GST and GST-tagged proteins. (C) 2013 Elsevier Inc. All rights reserved. |
| 英文摘要 | Novel GSH-AP (phenoxyl agarose coated gigaporous polystyrene, Agap-co-PSt) microspheres were successfully prepared by introducing GSH ligand into hydrophilic AP microspheres pre-activated with 1,4-butanediol diglycidyl ether. The gigaporous structure and chromatographic properties of GSH-AP medium were evaluated and compared with commercial GSH Sepharose FF (GSH-FF) medium. The macropores (100-500 nm) of gigaporous PSt microspheres were well maintained after coating with agarose and functionalized with GSH ligand. Hydrodynamic experiments showed that GSH-AP column had less backpressure and plate height than those of GSH-FF column at high flow velocity, which was beneficial for its use in high-speed chromatography. The presence of flow-through pores in GSH-AP microspheres also accelerated the mass transfer rate of biomolecules induced by convective flow, leading to high protein resolution and high dynamic binding capacity (DEC) of glutathione S-transferase (GST) at high flow velocity. High purity of GST and GST-tagged recombinant human interleukin-1 receptor antagonist (rhIL-1RA) were obtained from crude extract with an acceptable recovery yield within 1.5 min at a velocity up to 1400 cm/h. GSH-AP medium is promising for high-speed affinity chromatography for the purification of GST and GST-tagged proteins. (C) 2013 Elsevier Inc. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology |
| 关键词[WOS] | MICELLES SWELLING METHOD ; PERFUSION CHROMATOGRAPHY ; POLYSTYRENE MICROSPHERES ; PURIFICATION ; SEPARATION ; TAGS ; ADSORBENTS ; PARTICLES ; PHASE |
| 收录类别 | SCI |
| 原文出处 | |
| 语种 | 英语 |
| WOS记录号 | WOS:000332192900012 |
| 公开日期 | 2014-05-06 |
| 版本 | 出版稿 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/8150]  |
| 专题 | 过程工程研究所_研究所（批量导入） |
| 作者单位 | Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China |
| 推荐引用方式 GB/T 7714 | Huang, Yongdong,Zhang, Rongyue,Li, Juan,et al. A novel gigaporous GSH affinity medium for high-speed affinity chromatography of GST-tagged proteins[J]. PROTEIN EXPRESSION AND PURIFICATION,2014,95(1):84-91. |
| APA | Huang, Yongdong,Zhang, Rongyue,Li, Juan,Li, Qiang,Su, Zhiguo,&Ma, Guanghui.(2014).A novel gigaporous GSH affinity medium for high-speed affinity chromatography of GST-tagged proteins.PROTEIN EXPRESSION AND PURIFICATION,95(1),84-91. |
| MLA | Huang, Yongdong,et al."A novel gigaporous GSH affinity medium for high-speed affinity chromatography of GST-tagged proteins".PROTEIN EXPRESSION AND PURIFICATION 95.1(2014):84-91. |
入库方式： OAI收割
来源：过程工程研究所
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