Abstract
Expanded bed adsorption (EBA) was examined as the initial capture/purification step in the purification of monoclonal antibodies from Chinese hamster ovary (CHO) cultures. Two process alternatives each using EBA were compared to a conventional Protein A process without EBA. One alternative used Protein A affinity EBA followed by packed-bed cation and anion-exchange steps. The other alternative used cation-exchange EBA as the capture step followed by packed-bed Protein A and anion-exchange steps. The process using Protein A EBA produced comparable purity (host cell protein, DNA, Protein A, antibody aggregate) to the conventional process. However, the Protein A EBA column showed a significant decrease in dynamic capacity with a limited number of cycles. The process using cation EBA achieved comparable levels of host cell proteins (HCP) and DNA but not antibody aggregate or leached Protein A compared to the conventional process.
Similar content being viewed by others
References
Barnfield F, Hjorth R & Hammarstrom A (1994) Pilot scale recovery of recombinant anexin V from unclarified Escherichia coli homogenate using expanded bed adsorption. Biotech. Bioeng. 44: 922–929.
Erickson JC, Finch JD & Greene DC (1994) Direct capture of recombinant proteins from animal cell culture media using a fluidized bed adsorber. In: Berthold RE & Berthold W (eds.) Animal Cell Technology-Products for Today, Perspectives for Tomorrow (pp. 557–562) Butterworth & Heinemann, Oxford.
Fahrner RL, Blank GS & Zapata G. (1999) Expanded bed protein A affinity chromatography of a recombinant humanized monoclonal antibody: process development, operation, and comparison with a packed bed method J. Biotech. 75: 273–280.
Hansson M, Stahl S, Hjorth R, Uhlen M & Moks T (1994) Singlestep recovery of a secreted recombinant protein by expanded bed adsorption. Bio/Technology 12: 285–288.
Thommes J, Halfar M, Lenz S & Kula M.-R (1995) Purification of monoclonal antibodies from whole hybridoma fermentation broth by fluidized bed adsorption. Biotechnol. Bioeng. 45: 205–211.
Thommes J, Baker A, Halfar M, Karau A & Kula M.-R (1996) Isolation of monoclonal antibodies from cell containing hybridoma broth using a protein A coated adsorbent in expanded beds. J. Chromatogr. A 752: 111–122.
Zapata G (1999) Cation exchange expanded bed chromatography-one step combining initial harvesting, purification and concentration applicable to humanized IgG antibodies and antibody fragments. Presentation at BioTherapeutics '99: Evaluating Primary Recovery Technologies. Washington, DC, USA.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Blank, G., Zapata, G., Fahrner, R. et al. Expanded bed adsorption in the purification of monoclonal antibodies: a comparison of process alternatives. Bioseparation 10, 65–71 (2001). https://doi.org/10.1023/A:1012278022349
Issue Date:
DOI: https://doi.org/10.1023/A:1012278022349