Isolation of monoclonal antibodies from cell containing hybridoma broth using a protein A coated adsorbent in expanded beds.
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.
Thömmes
J
J
Institut für Enzymtechnologie, Heinrich-Heine Universität Düsseldorf, Jüllich, Germany.
Bader
A
A
Halfar
M
M
Karau
A
A
Kula
M R
MR
D000911:Antibodies, Monoclonal / Q000302:isolation & purification*; D006825:Hybridomas / Q000276:immunology*; D013205:Staphylococcal Protein A / Q000276:immunology*
D000911:Antibodies, Monoclonal; D013205:Staphylococcal Protein A