title: Enhancing IgG purification from serum albumin containing feedstock with hydrophobic charge-induction chromatography.
abstract: Hydrophobic charge-induction chromatography (HCIC) with 4-mercaptoethyl-pyridine (MEP) as the ligand is a novel technology for antibody purification, however, the separation selectivity still needs to be improved for the applications, especially for the impurity of serum albumin. In this study, with bovine serum immunoglobulin G (IgG) as the model, the purification of IgG from the serum albumin containing feedstock was developed with the commercial HCIC resin MEP HyperCel, focusing on the optimization of operation pH and salt addition. The adsorption isotherms of IgG and bovine serum albumin (BSA) were investigated at different pHs, and the binding and elution behaviors of two proteins in the column were also studied at varying pHs. In addition, the protein-ligand interactions were investigated with some additives in the buffer. It was found that the conditions of pH 6 with 0.1 M NaCl or pH 8 could be used to effectively remove BSA from the MEP resin without the influence on IgG adsorption. Two modes with control of loading or washing buffer were tested to enhance the purification of IgG from BSA containing feedstock, and the purity of IgG was improved to about 95% compared with 62.9% for the control. The results demonstrated that the control of loading pH or the addition of NaCl in the buffer might be an effective method to improve the purification of antibody with the HCIC process.
authors.affiliation: State Key Laboratory of Chemical Engineering, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou 310027, China.
authors.forename: Hong-Fei
authors.initials: HF
authors.lastname: Tong
authors.forename: Dong-Qiang
authors.initials: DQ
authors.lastname: Lin
authors.forename: Xiao-Ming
authors.initials: XM
authors.lastname: Yuan
authors.forename: Shan-Jing
authors.initials: SJ
authors.lastname: Yao
mesh_terms: D000327:Adsorption; D000818:Animals; D002417:Cattle; D002853:Chromatography, Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007074:Immunoglobulin G / Q000302:isolation & purification* / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D012965:Sodium Chloride; D012972:Sodium Hydroxide
chemical_list: D007074:Immunoglobulin G; D012710:Serum Albumin, Bovine; D012965:Sodium Chloride; D012972:Sodium Hydroxide