Impact of ionic strength on adsorption capacity of chromatographic particles employed in separation of monoclonal antibodies
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The influence of ionic strength on the adsorption capacity of seven commercial adsorbents used in downstream processing of monoclonal antibodies was examined. Affinity (MabSelect, Poros 50A High Capacity, ProSep-vA High Capacity), hydrophobic charge-induction (MEP HyperCel), and cation exchange adsorbents (FractoGel EMD SE Hicap (M), SP Sepharose Fast Flow, Ceramic HyperD F) were used to study the adsorption of polyclonal human immunoglobulin G at optimal pH values. The ionic strength, adjusted by sodium chloride concentrations in the range of 0–225 mM, strongly decreased the adsorption capacity of the cation exchangers. Equilibrium data were described in the form of the dependence of the ratio of protein concentrations in the solid and liquid phases on the total concentration of cation counter ions. They were successfully fitted and interpreted through a stoichiometric ion-exchange model.
Keywordsmonoclonal antibody adsorption capacity protein A chromatography cation-exchange chromatography hydrophobic charge-induction chromatography ionic strength pH effect stoichiometric model
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- Gramblička, M., Tóthová, D., Antošová, M., & Polakovič, M. (2008). Influence of pH on adsorption of human immunoglobulin gamma, human serum albumin and horse myoglobin by commercial chromatographic materials designed for downstream processing of monoclonal antibodies. Acta Chimica Slovaca, 1(1) 85–94.Google Scholar
- Hahn, R., Shimahara, K., Steindl, F., & Jungbauer, A. (2006). Comparison of protein A affinity sorbents III. Life time study. Journal of Chromatography A, 1102, 224–231. DOI: 10.1016/j.chroma.2005.10.083.Google Scholar
- Necina, R., Amatschek, K., & Jungbauer, A. (1998). Capture of human monoclonal antibodies from cell culture supernatant by ion exchange media exhibiting high charge density. Biotechnology and Bioengineering, 60, 689–698. DOI: 10.1002/(SICI)1097-0290(19981220)60:6〈689::AID-BIT6〉3.0.CO;2-M.CrossRefGoogle Scholar
- Schwartz, W., Judd, D., Wysocki, M., Guerrier, L., Birck-Wilson, E., & Boschetti, E. (2001). Comparison of hydrophobic charge induction chromatography with affinity chromatography on protein A for harvest and purification of antibodies. Journal of Chromatography A, 908, 251–263. DOI: 10.1016/S0021-9673(00)01013-X.CrossRefGoogle Scholar
- Smith, A. W. (1948). Elements of physics (5th ed.). New York, NY, USA: McGraw-Hill.Google Scholar
- Staby, A., Sand, M.-B., Hansen, R. G., Jacobsen, J. H., Andersen, L. A., Gerstenberg, M., Bruus, U. K., & Holm Jensen, I. (2005). Comparison of chromatographic ion-exchange resins: IV. Strong and weak cation-exchange resins and heparin resins. Journal of Chromatography A, 1069, 65–77. DOI: 10.1016/j.chroma.2004.11.094.CrossRefGoogle Scholar
- Stone, M. C., Tao, Y., & Carta, G. (2009). Protein adsorption and transport in agarose and dextran-grafted agarose media for ion exchange chromatography: Effect of ionic strength and protein characteristics. Journal of Chromatography A, 1216, 4465–4474. DOI: 10.1016/j.chroma.2009.03.044.CrossRefGoogle Scholar
- Tatárová, I., Gramblička, M., Antošová, M., & Polakovič, M. (2008). Characterization of pore structure of chromatographic adsorbents employed in separation of monoclonal antibodies using size-exclusion techniques. Journal of Chromatography A, 1193, 129–135. DOI: 10.1016/j.chroma.2008.04.023.CrossRefGoogle Scholar