title: Preparation and characterization of a thermoresponsive gigaporous medium for high-speed protein chromatography.
abstract: A high-speed thermoresponsive medium was developed by grafting poly(N-isopropylacrylamide-co-butyl methacrylate) (P(NIPAM-co-BMA)) brushes onto gigaporous polystyrene (PS) microspheres via surface-initiated atom transfer radical polymerization (ATRP) technique, which has strong mechanical strength, good chemical stability and high mass transfer rate for biomacromolecules. The gigaporous structure, surface chemical composition, static protein adsorption, and thermoresponsive chromatographic properties of prepared medium (PS-P(NIPAM-co-BMA)) were characterized in detail. Results showed that the PS microspheres were successfully grafted with P(NIPAM-co-BMA) brushes and that the gigaporous structure was robustly maintained. After grafting, the nonspecific adsorption of proteins on PS microspheres was greatly reduced. A column packed with PS-P(NIPAM-co-BMA) exhibited low backpressure and significant thermo-responsibility. By simply changing the column temperature, it was able to separate three model proteins at the mobile phase velocity up to 2167 cm h(-1). In conclusion, the thermoresponsive polymer brushes grafted gigaporous PS microspheres prepared by ATRP are very promising in 'green' high-speed preparative protein chromatography.
authors.affiliation: State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China. Electronic address: jbqu@upc.edu.cn.
authors.forename: Jian-Bo
authors.initials: JB
authors.lastname: Qu
authors.affiliation: State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China.
authors.forename: Yan-Li
authors.initials: YL
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China.
authors.forename: Guan-Sheng
authors.initials: GS
authors.lastname: Huan
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, PR China.
authors.forename: Wei-Qing
authors.initials: WQ
authors.lastname: Zhou
authors.affiliation: State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China.
authors.forename: Jian-Guo
authors.initials: JG
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China.
authors.forename: Hu
authors.initials: H
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Heavy Oil Processing, Center for Bioengineering and Biotechnology, China University of Petroleum (East China), Qingdao 266580, PR China. Electronic address: zhangxy@upc.edu.cn.
authors.forename: Xiao-Yun
authors.initials: XY
authors.lastname: Zhang
mesh_terms: D002853:Chromatography, Liquid / Q000295:instrumentation* / Q000379:methods; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification; D008863:Microspheres; D011109:Polymethacrylic Acids / Q000737:chemistry; D011137:Polystyrenes / Q000737:chemistry; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000302:isolation & purification; D013696:Temperature
chemical_list: D011109:Polymethacrylic Acids; D011137:Polystyrenes; D011506:Proteins; C039217:fluorescein isothiocyanate bovine serum albumin; C084450:poly(N-isopropylacrylamide-co-butyl methacrylate); D012710:Serum Albumin, Bovine; D016650:Fluorescein-5-isothiocyanate
keywords: Atom transfer radical polymerization; Gigaporous polystyrene microspheres; N-isopropylacrylamide; Protein chromatography; Thermoresponsive polymer brushes