title: Enhanced binding by dextran-grafting to Protein A affinity chromatographic media.
abstract: Dextran-grafted Protein A affinity chromatographic medium was prepared by grafting dextran to agarose-based matrix, followed by epoxy-activation and Protein A coupling site-directed to sulfhydryl groups of cysteine molecules. An enhancement of both the binding performance and the stability was achieved for this dextran-grafted Protein A chromatographic medium. Its dynamic binding capacity was 61 mg immunoglobulin G/mL suction-dried gel, increased by 24% compared with that of the non-grafted medium. The binding capacity of dextran-grafted medium decreased about 7% after 40 cleaning-in-place cycles, much lower than that of the non-grafted medium as decreased about 15%. Confocal laser scanning microscopy results showed that immunoglobulin G was bound to both the outside and the inside of dextran-grafted medium faster than that of non-grafted one. Atomic force microscopy showed that this dextran-grafted Protein A medium had much rougher surface with a vertical coordinate range of ±80 nm, while that of non-grafted one was ±10 nm. Grafted dextran provided a more stereo surface morphology and immunoglobulin G molecules were more easily to be bound. This high-performance dextran-grafted Protein A affinity chromatographic medium has promising applications in large-scale antibody purification.
authors.affiliation: State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.
authors.forename: Lan
authors.initials: L
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.|School of Chemical and Environmental Engineering, China University of Mining and Technology, Beijing, P.R. China.
authors.forename: Kai
authors.initials: K
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.
authors.forename: Yongdong
authors.initials: Y
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Li
authors.affiliation: State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.
authors.forename: Xiunan
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Chemical Engineering, Beijing Institute of Petro-chemical Technology, Beijing, P.R. China.
authors.forename: Rongyue
authors.initials: R
authors.lastname: Zhang
authors.affiliation: State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.
authors.forename: Zhiguo
authors.initials: Z
authors.lastname: Su
authors.affiliation: School of Chemical and Environmental Engineering, China University of Mining and Technology, Beijing, P.R. China.
authors.forename: Qibao
authors.initials: Q
authors.lastname: Wang
authors.affiliation: State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing, P.R. China.|Jiangsu National Synergetic Innovation, Center for Advance Materials (SICAM), Nanjing, P.R. China.
authors.forename: Guanghui
authors.initials: G
authors.lastname: Ma
mesh_terms: D000327:Adsorption; D002846:Chromatography, Affinity; D003911:Dextrans / Q000737:chemistry*; D007074:Immunoglobulin G / Q000378:metabolism; D011485:Protein Binding; D012685:Sepharose / Q000737:chemistry; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000378:metabolism*
chemical_list: D003911:Dextrans; D007074:Immunoglobulin G; D013205:Staphylococcal Protein A; D012685:Sepharose
keywords: Protein A; affinity chromatography; antibody purification; dextran; grafting