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Impact of ionic strength on adsorption capacity of chromatographic particles employed in separation of monoclonal antibodies

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Abstract

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.

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References

  • Barrande, M., Beurroies, I., Denoyel, R., Tatárová, I., Gramblička, M., Polakovič, M., Joehnck, M., & Schulte, M. (2009). Characterisation of porous materials for bioseparation. Journal of Chromatography A, 1216, 6906–6916. DOI: 10.1016/j.chroma.2009.07.075.

    Article  CAS  Google Scholar 

  • Boschetti, E. (2002). Antibody separation by hydrophobic charge induction chromatography. Trends in Biotechnology, 20, 333–337. DOI: 10.1016/S0167-7799(02)01980-7.

    Article  CAS  Google Scholar 

  • Burton, S. C., & Harding, D. R. K. (1998). Hydrophobic charge induction chromatography: salt independent protein adsorption and facile elution with aqueous buffers. Journal of Chromatography A, 814, 71–81. DOI: 10.1016/S0021-9673(98)00436-1.

    Article  CAS  Google Scholar 

  • Carter-Franklin, N., Victa, C., McDonald, P., & Fahrner, R. (2007). Fragments of protein A eluted during protein A affinity chromatography. Journal of Chromatography A, 1163, 105–111. DOI: 10.1016/j.chroma.2007.06.012.

    Article  CAS  Google Scholar 

  • Chen, J., Tetrault, J., & Ley, A. (2008). Comparison of standard and new generation hydrophobic interaction chromatography resins in the monoclonal antibody purification process. Journal of Chromatography A, 1177, 272–281. DOI: 10.1016/j.chroma.2007.07.083.

    Article  CAS  Google Scholar 

  • Denton, G., Murray, A., Price, M. R., & Levison, P. R. (2001). Direct isolation of monoclonal antibodies from tissue culture supernatant using the cation-exchange cellulose Express-Ion S. Journal of Chromatography A, 908, 223–234. DOI: 10.1016/S0021-9673(00)00834-7.

    Article  CAS  Google Scholar 

  • Faude, A., Zacher, D., Müller, E., & Böttinger, H. (2007). Fast determination of conditions for maximum dynamic capacity in cation-exchange chromatography of human monoclonal antibodies. Journal of Chromatography A, 1161, 29–35 DOI: 10.1016/j.chroma.2007.03.114.

    Article  CAS  Google Scholar 

  • Follman, D. K., & Fahrner, R. L. (2004). Factorial screening of antibody purification processes using three chromatography steps without protein A. Journal of Chromatography A, 1024, 79–85. DOI: 10.1016/j.chroma.2003.10.060.

    Article  CAS  Google Scholar 

  • Forrer, N., Butté, A., & Morbidelli, M. (2008). Chromatographic behavior of a polyclonal antibody mixture on a strong cation exchanger column. Part I: Adsorption characterization Journal of Chromatography A, 1214, 59–70. DOI: 10.1016/j.chroma.2008.10.048.

    Article  CAS  Google Scholar 

  • Ghose, S., Hubbard, B., & Cramer, S. M. (2006). Evaluation and comparison of alternatives to Protein A chromatography: Mimetic and hydrophobic charge induction chromatographic stationary phases. Journal of Chromatography A, 1122, 144–152. DOI: 10.1016/j.chroma.2006.04.083.

    Article  CAS  Google Scholar 

  • 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 

  • Guerrier, L., Flayeux, I., & Boschetti, E. (2001). A dual-mode approach to the selective separation of antibodies and their fragments. Journal of Chromatography B, 755, 37–46. DOI: 10.1016/S0378-4347(00)00598-3.

    Article  CAS  Google Scholar 

  • Hahn, R., Schlegel, R., & Jungbauer, A. (2003). Comparison of protein A affinity sorbents. Journal of Chromatography B, 790, 35–51. DOI: 10.1016/S1570-0232(03)00092-8.

    Article  CAS  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.

    CAS  Google Scholar 

  • Hober, S., Nord, K., & Linhult, M. (2007). Protein A chromatography for antibody purification. Journal of Chromatography B, 848, 40–47. DOI: 10.1016/j.jchromb.2006.09. 030.

    Article  CAS  Google Scholar 

  • Huse, K., Böhme, H.-J., & Scholz, G. H. (2002). Purification of antibodies by affinity chromatography. Journal of Biochemical and Biophysical Methods, 51, 217–231. DOI: 10.1016/S0165-022X(02)00017-9.

    Article  CAS  Google Scholar 

  • Jungbauer, A. (2005). Chromatographic media for bioseparation. Journal of Chromatography A, 1065, 3–12. DOI: 10.1016/j.chroma.2004.08.162.

    Article  CAS  Google Scholar 

  • Low, D., O’Leary, R., & Pujar, N. S. (2007). Future of antibody purification. Journal of Chromatography B, 848, 48–63. DOI: 10.1016/j.jchromb.2006.10.033.

    Article  CAS  Google Scholar 

  • Mollerup, J. M. (2006). Applied thermodynamics: A new frontier for biotechnology. Fluid Phase Equilibria, 241, 205–215. DOI: 10.1016/j.fluid.2005.12.037.

    Article  CAS  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.

    Article  CAS  Google Scholar 

  • Okay, O. (2000). Macroporous copolymer networks. Progress in Polymer Science, 25, 711–779. DOI: 10.1016/S0079-6700(00)00015-0.

    Article  CAS  Google Scholar 

  • Roque, A. C. A., Silva, C. S. O., & Taipa, M. Á. (2007). Affinitybased methodologies and ligands for antibody purification: Advances and perspectives. Journal of Chromatography A, 1160, 44–55. DOI: 10.1016/j.chroma.2007.05.109.

    Article  CAS  Google 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.

    Article  CAS  Google Scholar 

  • Shukla, A. A., Hubbard, B., Tressel, T., Guhan, S., & Low, D. (2007). Downstream processing of monoclonal antibodies—Application of platform approaches. Journal of Chromatography B, 848, 28–39. DOI: 10.1016/j.jchromb.2006.09.026.

    Article  CAS  Google Scholar 

  • Smith, A. W. (1948). Elements of physics (5th ed.). New York, NY, USA: McGraw-Hill.

    Google Scholar 

  • Staby, A., Jacobsen, J. H., Hansen, R. G., Bruus, U. K., & Holm Jensen, I. (2006). Comparison of chromatographic ion-exchange resins: V. Strong and weak cation-exchange resins. Journal of Chromatography A, 1118, 168–179. DOI: 10.1016/j.chroma.2006.03.116.

    Article  CAS  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.

    Article  CAS  Google 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.

    Article  CAS  Google 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.

    Article  Google Scholar 

  • Tugcu, N., Bae, S. S., Moore, J. A., & Cramer, S. M. (2002). Stationary phase effects on the dynamic affinity of lowmolecular-mass displacers. Journal of Chromatography A, 954, 127–135. DOI: 10.1016/S0021-9673(02)00164-4.

    Article  CAS  Google Scholar 

  • Zhu-Shimoni, J., Gunawan, F., Thomas, A., Vanderlaan, M., & Stults, J. (2009). Trace level analysis of leached Protein A in bioprocess samples without interference from the large excess of rhMAb IgG. Journal of Immunological Methods, 341, 59–67. DOI: 10.1016/j.jim.2008.10.015.

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Chemical and Biochemical Engineering, Faculty of Chemical and Food Technology, Slovak University of Technology, Radlinského 9, 812 37, Bratislava, Slovak Republic

    Katarzyna Wrzosek, Michal Gramblička, Darina Tóthová, Monika Antošová & Milan Polakovič

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  1. Katarzyna Wrzosek
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  2. Michal Gramblička
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  3. Darina Tóthová
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  4. Monika Antošová
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  5. Milan Polakovič
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Correspondence to Milan Polakovič.

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Wrzosek, K., Gramblička, M., Tóthová, D. et al. Impact of ionic strength on adsorption capacity of chromatographic particles employed in separation of monoclonal antibodies. Chem. Pap. 64, 461–468 (2010). https://doi.org/10.2478/s11696-010-0019-5

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  • DOI: https://doi.org/10.2478/s11696-010-0019-5

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