title: Functional gigaporous polystyrene microspheres facilitating separation of poly(ethylene glycol)-protein conjugate.
abstract: A novel sulfopropyl gigaporous polystyrene (SP-GP) microsphere enhancing the separation of poly(ethylene glycol)-protein (PEGylated protein) was first presented. The SP-GP microspheres were successfully prepared by introducing sulfopropyl groups into agarose-coated gigaporous polystyrene microspheres and used as chromatography media. Compared with a commercial medium, SP-GP microspheres exhibited improved column efficiency and reduced backpressure with increasing flow velocity, which could ensure its use in high-speed chromatography. Furthermore, a higher protein recovery and purity of the PEGylated protein could be obtained, even when SP-GP was applied at a flow velocity of 1224 cm h(-1). Additionally, the dynamic binding capacity (DBC) of SP-GP was significantly improved, which was higher than 10 mg mL(-1) medium even at a flow velocity of 306 cm h(-1). Further investigation using a laser scanning confocal microscope (LSCM) demonstrated that the static adsorption equilibrium of the PEGylated protein on SP-GP could be completed in 5 min, whereas a much longer period (ca. 60 min) was required for the commercial medium, indicating that the mass transfer of SP-GP was much faster with the gigaporous structure. All of these results strongly support that our developed SP-GP could serve as a promising cation exchange chromatography resin for high-speed separation, especially for biomolecules of high molecular weight.
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, PR China.
authors.forename: Yanqin
authors.initials: Y
authors.lastname: Zhai
authors.forename: Weiqing
authors.initials: W
authors.lastname: Zhou
authors.forename: Wei
authors.initials: W
authors.lastname: Wei
authors.forename: Jianbo
authors.initials: J
authors.lastname: Qu
authors.forename: Jiandu
authors.initials: J
authors.lastname: Lei
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Su
authors.forename: Guanghui
authors.initials: G
authors.lastname: Ma
mesh_terms: D002850:Chromatography, Gel; D016179:Granulocyte Colony-Stimulating Factor / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D006801:Humans; D008863:Microspheres; D011092:Polyethylene Glycols / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011137:Polystyrenes / Q000737:chemistry*; D016062:Porosity; D011506:Proteins / Q000737:chemistry* / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D011137:Polystyrenes; D011506:Proteins; D011994:Recombinant Proteins; C486981:pegylated granulocyte colony-stimulating factor, human; D016179:Granulocyte Colony-Stimulating Factor; D011092:Polyethylene Glycols