title: Efficient fabrication of high-capacity immobilized metal ion affinity chromatographic media: The role of the dextran-grafting process and its manipulation.
abstract: Novel high-capacity Ni(2+) immobilized metal ion affinity chromatographic media were prepared through the dextran-grafting process. Dextran was grafted to an allyl-activated agarose-based matrix followed by functionalization for the immobilized metal ion affinity chromatographic media. With elaborate regulation of the allylation degree, dextran was completely or partly grafted to agarose microspheres, namely, completely dextran-grafted agarose microspheres and partly dextran-grafted ones, respectively. Confocal laser scanning microscope results demonstrated that a good adjustment of dextran-grafting degree was achieved, and dextran was distributed uniformly in whole completely dextran-grafted microspheres, while just distributed around the outside of the partly dextran-grafted ones. Flow hydrodynamic properties were improved greatly after the dextran-grafting process, and the flow velocity increased by about 30% compared with that of a commercial chromatographic medium (Ni Sepharose FF). A significant improvement of protein binding performance was also achieved by the dextran-grafting process, and partly dextran-grafted Ni(2+) chelating medium had a maximum binding capacity for His-tagged lactate dehydrogenase about 2.5 times higher than that of Ni Sepharose FF. The results indicated that this novel chromatographic medium is promising for applications in high-efficiency and large-scale protein purification.
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, PR China.
authors.forename: Lan
authors.initials: L
authors.lastname: Zhao
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, PR China.|Hebei Normal University of Science & Technology, Qinhuangdao, Hebei, PR China.
authors.forename: Jingfei
authors.initials: J
authors.lastname: Zhang
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, PR China.
authors.forename: Yongdong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, PR China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Chemical Engineering, Beijing Institute of Petro-chemical Technology, Beijing, PR China.
authors.forename: Rongyue
authors.initials: R
authors.lastname: Zhang
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, PR China.|School of Chemical & Environmental Engineering, China University of Mining & Technology, Beijing, PR China.
authors.forename: Kai
authors.initials: K
authors.lastname: Zhu
authors.affiliation: Department of Chemical Engineering, Beijing Institute of Petro-chemical Technology, Beijing, PR China.
authors.forename: Jia
authors.initials: J
authors.lastname: Suo
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, PR China.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Su
authors.affiliation: Hebei Normal University of Science & Technology, Qinhuangdao, Hebei, PR China.
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, PR China.
authors.forename: Guanghui
authors.initials: G
authors.lastname: Ma
mesh_terms: D002614:Chelating Agents / Q000138:chemical synthesis / Q000737:chemistry*; D002846:Chromatography, Affinity / Q000379:methods*; D003911:Dextrans / Q000737:chemistry*; D057446:Hydrodynamics; D007477:Ions / Q000737:chemistry; D009532:Nickel / Q000737:chemistry*; D010316:Particle Size; D013499:Surface Properties
chemical_list: D002614:Chelating Agents; D003911:Dextrans; D007477:Ions; D009532:Nickel
keywords: Dextran; Immobilized metal ion affinity chromatography; Nickel; Protein purification