Arginine as an eluent for automated on-line Protein A/size exclusion chromatographic analysis of monoclonal antibody aggregates in cell culture

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
Sean Wang, Anil Raghani

Abstract

An automated two-dimensional method using Protein A chromatography followed by size exclusion HPLC was developed for analysis of aggregates of monoclonal antibodies in mammalian cell culture samples. The method development was intended to address the analysis of IgG2 monoclonal antibody products that are particularly prone to aggregation at pH<3.5. An aqueous solution of sodium phosphate at pH 4.3 containing arginine was demonstrated in this work as an excellent eluent for Protein A chromatography. In addition, the arginine solution is a compatible mobile phase for the analysis of these samples by size exclusion HPLC, separating aggregates from the monomer of the monoclonal antibody. The effect of arginine concentration in the eluent on parameters such as protein recovery from Protein A chromatography and resolution of aggregates from the monomer are reported. The developed method was shown to provide accuracy of reported aggregates greater than 98%, and intermediate precision of 4.4% RSD. The method limit of quantitation for aggregates was determined to be 0.1%. Application of the method is demonstrated for analysis of aggregates in cell culture samples to aid in the development of cell culture conditions for the production of...Continue Reading

References

Aug 30, 1996·Journal of Chromatography. a·R D Ricker, L A Sandoval
Jul 14, 2004·Protein Expression and Purification·Tsutomu ArakawaDaisuke Ejima
Nov 1, 2005·Journal of Chromatography. a·Daisuke EjimaKouhei Tsumoto
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Jul 30, 2011·Neurological Sciences : Official Journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology·Luisa ChiappariniLaura Grazia Valentini

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Citations

Aug 19, 2015·Analytical Chemistry·Amy FarrellJonathan Bones
Jul 19, 2014·Current Opinion in Biotechnology·Daniel A M PaisAna P Teixeira
May 27, 2014·Biochimica Et Biophysica Acta·Tsutomu ArakawaDaisuke Ejima

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