title: Arginine as an eluent overcomes the hindrance of monoclonal antibody quantification by dextran sulfate in protein A affinity chromatography.
abstract: Analytical chromatography using protein A affinity columns was employed for the fast and simple quantitative analysis of monoclonal antibodies (mAb) from suspension cultures of recombinant Chinese hamster ovary (rCHO) cells. Reliable results could not be obtained from analysis of rCHO cell culture supernatants containing dextran sulfate using elution buffers such as phosphate, glycine, or MgCl2 . These problems increased as the number of analysis and the concentration of dextran sulfate in samples increased. Arginine was identified as an alternative eluent to overcome the hindrance by dextran sulfate. When the samples contain dextran sulfate up to 100 mg/L, the elution buffer containing 0.6-1.0 M arginine at pH 3.0-3.8 is useful for the effective analysis. Reproducible results in the mAb quantification could be obtained by this developed arginine elution buffer from rCHO cell culture supernatants containing dextran sulfate.
authors.affiliation: Dept. of Chemical Engineering, Hanyang University, Seoul, 133-791, Korea.
authors.forename: Bong Gyun
authors.initials: BG
authors.lastname: Kim
authors.affiliation: Dept. of Chemical Engineering, Hanyang University, Seoul, 133-791, Korea.
authors.forename: Hong Woo
authors.initials: HW
authors.lastname: Park
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D001120:Arginine / Q000737:chemistry*; D016466:CHO Cells; D002846:Chromatography, Affinity / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D016264:Dextran Sulfate / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D015636:Magnesium Chloride; D013205:Staphylococcal Protein A / Q000737:chemistry* / Q000378:metabolism
chemical_list: D000911:Antibodies, Monoclonal; D013205:Staphylococcal Protein A; D015636:Magnesium Chloride; D016264:Dextran Sulfate; D001120:Arginine
keywords: antibody; arginine; dextran sulfate; protein A