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A novel gigaporous GSH affinity medium for high-speed affinity chromatography of GST-tagged proteins.

Abstract：

: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 (100-500nm) 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 (DBC) 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 1.5min at a velocity up to 1400cm/h. GSH-AP medium is promising for high-speed affinity chromatography for the purification of GST and GST-tagged proteins.

journal_name

Protein Expr Purif

journal_title

Protein expression and purification

authors

Huang Y,Zhang R,Li J,Li Q,Su Z,Ma G

doi

10.1016/j.pep.2013.11.004

subject

Has Abstract

pub_date

2014-03-01 00:00:00

pages

84-91

eissn

1046-5928

issn

1096-0279

pii

S1046-5928(13)00241-6

journal_volume

95

pub_type

杂志文章

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