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Arginine as an eluent overcomes the hindrance of monoclonal antibody quantification by dextran sulfate in protein A affinity chromatography
Author(s) -
Kim Bong Gyun,
Park Hong Woo
Publication year - 2015
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.2164
Subject(s) - chromatography , dextran , chemistry , chinese hamster ovary cell , elution , column chromatography , arginine , monoclonal antibody , affinity chromatography , quantitative analysis (chemistry) , sulfate , biochemistry , antibody , amino acid , biology , enzyme , receptor , immunology , organic chemistry
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 MgCl 2 . 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. © 2015 American Institute of Chemical Engineers Biotechnol. Prog. , 31:1536–1541, 2015