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A novel gigaporous GSH affinity medium for high-speed affinity chromatography of GST-tagged proteins
Alternative TitleProtein Expr. Purif.
Huang, Yongdong; Zhang, Rongyue; Li, Juan; Li, Qiang; Su, Zhiguo; Ma, Guanghui
2014-03-01
Source PublicationPROTEIN EXPRESSION AND PURIFICATION
ISSN1046-5928
Volume95Issue:1Pages:84-91
AbstractNovel 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.; 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.
KeywordGsh Affinity Chromatography Gigaporous Polystyrene Microspheres High-speed Chromatography Gst-tagged Proteins
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine
DOI10.1016/j.pep.2013.11.004
URL查看原文
Indexed BySCI
Language英语
WOS KeywordMICELLES SWELLING METHOD ; PERFUSION CHROMATOGRAPHY ; POLYSTYRENE MICROSPHERES ; PURIFICATION ; SEPARATION ; TAGS ; ADSORBENTS ; PARTICLES ; PHASE
WOS Research AreaBiochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS SubjectBiochemical Research Methods ; Biochemistry & Molecular Biology ; Biotechnology & Applied Microbiology
WOS IDWOS:000332192900012
Citation statistics
Cited Times:3[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/8150
Collection研究所(批量导入)
AffiliationChinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China
Recommended Citation
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.
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