title: A novel matrix derivatized from hydrophilic gigaporous polystyrene-based microspheres for high-speed immobilized-metal affinity chromatography.
abstract: Agarose coated gigaporous polystyrene microspheres were evaluated as a novel matrix for immobilized-metal affinity chromatography (IMAC). With four steps, nickel ions were successfully immobilized on the microspheres. The gigaporous structure and chromatographic properties of IMAC medium were characterized. A column packed with the matrix showed low column backpressure and high column efficiency at high flow velocity. Furthermore, this matrix was used for purifying superoxide dismutase (SOD), which was expressed in Escherichia coli (E. coli) in submerged fermentation, on an Äkta purifier 100 system under different flow velocities. The purity of the SOD from this one-step purification was 79% and the recovery yield was about 89.6% under the superficial flow velocity of 3251 cm/h. In conclusion, all the results suggested that the gigaporous matrix has considerable advantages for high-speed immobilized-metal affinity chromatography.
authors: Center for Bioengineering and Biotechnology, China University of Petroleum (East China), QingDao, PR China. jbqu@upc.edu.cn
authors: Jian-Bo
authors: JB
authors: Qu
authors: Yong-Dong
authors: YD
authors: Huang
authors: Guang-Lun
authors: GL
authors: Jing
authors: Jian-Guo
authors: JG
authors: Liu
authors: Wei-Qing
authors: WQ
authors: Zhou
authors: Hu
authors: H
authors: Zhu
authors: Jian-Ren
authors: JR
authors: Lu
mesh_terms: D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D004586:Electrophoresis; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D057927:Hydrophobic and Hydrophilic Interactions; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D009532:Nickel / Q000737:chemistry*; D010316:Particle Size; D011137:Polystyrenes / Q000737:chemistry*; D016062:Porosity; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification; D013482:Superoxide Dismutase / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification
chemical_list: D011137:Polystyrenes; D011993:Recombinant Fusion Proteins; D009532:Nickel; D013482:Superoxide Dismutase