Isolation of monoclonal antibodies from cell containing hybridoma broth using a protein A coated adsorbent in expanded beds

• Published in: Journal of Chromatography A 1996-11, Vol.752 (1), p.111-122
• Main Authors: Thömmes, Jörg, Bader, Andrea, Halfar, Markus, Karau, Andreas, Kula, Maria-Regina
• Format: Article
• Language: English
• Subjects: Antibodies, Monoclonal - isolation & purification; Biological and medical sciences; Biotechnology; Expanded bed adsorption; Fluidized bed adsorption; Fundamental and applied biological sciences. Psychology; Health. Pharmaceutical industry; Hybridomas - immunology; Immunoadsorbents; Industrial applications and implications. Economical aspects; Monoclonal antibodies; Production of active biomolecules; Protein A; Staphylococcal Protein A - immunology
• ISSN: 0021-9673
• DOI: 10.1016/S0021-9673(96)00504-3

A novel expanded bed adsorbent carrying a recombinant protein A ligand was used for the isolation of monoclonal antibodies from cell containing hybridoma fermentation broth. The untreated effluent from a continuous hybridoma cultivation was applied to the stable expanded adsorbent (Streamline rProteinA), which proved to have a high capacity for the MAb studied (14 mg MAb per ml of adsorbent). A clarified and highly concentrated (up to 50 fold) eluate of high purity was obtained. A scale up of the MAb purification is demonstrated from lab scale (250 mg MAb per purification cycle) to a small pilot scale (2 g MAb per cycle). Low product concentration in the broth in combination with the high capacity of the adsorbent caused long sample application cycles (10–11 h). Experimental problems arising from these long cycle times are discussed with regard to a large scale application of the method.