title: Protein adsorption behavior and immunoglobulin separation with a mixed-mode resin based on p-aminohippuric acid.
abstract: p-Aminohippuric acid is a newly developed ligand for mixed-mode chromatography with a commercial resin name of Nuvia cPrime. In this study, bovine immunoglobulin G and bovine serum albumin were used as two model proteins, and the adsorption isotherms with Nuvia cPrime were investigated under different pH and salt concentrations. The results showed that pH had a strong but different influence on the adsorption of these two proteins. The adsorption capacity for bovine immunoglobulin G and BSA was 170.4 and 28.1 mg/g at pH 6.0, respectively. Different salts also showed varying effects on the protein adsorption. Moreover, the adsorption and elution behaviors of the two proteins in a column were determined under varying pH and salt concentrations. An optimized process showed that feedstock loaded under pH 6.0 with 0.8 M (NH4)2SO4 and eluted under pH 8.0 with 1.0 M NaCl could effectively purify bovine immunoglobulin G from feedstock containing BSA. The purity of bovine immunoglobulin G could reach 99.8% and the recovery was 92.7%. The results demonstrated that the control of pH and salt addition during the loading and elution processes were two key factors in improving separation efficiency with Nuvia cPrime resin.
authors.affiliation: Key Laboratory of Biomass Chemical Engineering of Ministry of Education, Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Yan
authors.forename: Qi-Lei
authors.initials: QL
authors.lastname: Zhang
authors.forename: Dong-Qiang
authors.initials: DQ
authors.lastname: Lin
authors.forename: Shan-Jing
authors.initials: SJ
authors.lastname: Yao
mesh_terms: D000327:Adsorption; D000818:Animals; D002417:Cattle; D006863:Hydrogen-Ion Concentration; D007074:Immunoglobulin G / Q000737:chemistry* / Q000302:isolation & purification*; D012492:Salts / Q000737:chemistry; D012710:Serum Albumin, Bovine / Q000737:chemistry* / Q000302:isolation & purification*; D013499:Surface Properties; D010130:p-Aminohippuric Acid / Q000737:chemistry*
chemical_list: D007074:Immunoglobulin G; D012492:Salts; D012710:Serum Albumin, Bovine; D010130:p-Aminohippuric Acid
keywords: Adsorption; Bovine immunoglobulin G; Bovine serum albumin; Mixed-mode chromatography; Purification