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. © 2015 American Institute of Chemical Engineers Biotechnol. Prog., 31:1536–1541, 2015

A genome‐scale nutrient minimization forecast algorithm for controlling essential amino acid levels in CHO cell cultures

“…Solution of 50 mM of sodium phosphate with 150 mM sodium chloride at pH = 7.2 was prepared as binding buffer. Due to the high concentration of dextran sulfate presented in culture, 0.7 M of L-arginine solution at pH 3.0 (adjusted with hydrochloric acid) was used as elution buffer as suggested in (Kim & Park, 2015). The running flowrate is constant at 2.0 ml/min with a stepwise elution gradient.…”