title: Preparation and evaluation of dextran-grafted agarose resin for hydrophobic charge-induction chromatography.
abstract: Hydrophobic charge-induction chromatography (HCIC) is a new and effective technology for antibody separation. In the present work, HCIC resin MMI-B-XL was prepared with dextran-grafted agarose gel as the matrix and 2-mercapto-1-methyl-imidazole (MMI) as the functional ligand. The preparation procedures were optimized, and the maximum ligand density could reach as high as 200 μmol/g gel. The adsorption isotherms and kinetics on new resins were investigated with human immunoglobulin G (hIgG) as the model protein, which were compared with non-grafted HCIC resin MMI-B-6FF. It was found that the saturated adsorption capacity (Qm) increased with the increase of ligand density for MMI-B-XL. Moreover, the effective diffusivity (De) could be dramatically enhanced with the increase of ligand density for MMI-B-XL, and the De for MMI-B-XL with the ligand density of 200 μmol/g gel was 18-40 times higher than that for MMI-B-6FF. The breakthrough experiments indicated that new resins with the ligand density of 200 μmol/g gel could be used for high superficial velocity and high dynamic adsorption could be obtained. The results indicated that dextran-grafted layer on the resin could increase the ligand density, enhance the mass transport in the pore, and improve the dynamic adsorption at high velocity, which showed a potential application for large-scale antibody purification.
authors.affiliation: State Key Laboratory of Chemical Engineering, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Tao
authors.initials: T
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Chemical Engineering, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China. Electronic address: lindq@zju.edu.cn.
authors.forename: Dong-Qiang
authors.initials: DQ
authors.lastname: Lin
authors.affiliation: State Key Laboratory of Chemical Engineering, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Hui-Li
authors.initials: HL
authors.lastname: Lu
authors.affiliation: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China; Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin 300072, China.
authors.forename: Shan-Jing
authors.initials: SJ
authors.lastname: Yao
mesh_terms: D000327:Adsorption; D002853:Chromatography, Liquid / Q000295:instrumentation*; D003911:Dextrans / Q000737:chemistry*; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000737:chemistry / Q000302:isolation & purification; D007700:Kinetics; D008024:Ligands; D012685:Sepharose / Q000737:chemistry*
chemical_list: D003911:Dextrans; D007074:Immunoglobulin G; D008024:Ligands; D012685:Sepharose
keywords: Adsorption capacity; Adsorption kinetics; Dextran-grafted agarose resin; Human immunoglobulin G; Hydrophobic charge-induction chromatography; Ligand density