Purification of monoclonal antibodies from whole hybridoma fermentation broth by fluidized bed adsorption

To achieve the coarse purification of a monoclonal antibody from whole hybridoma fermentation broth a fluidized bed cation exchange process was used. The procedure consisted of application of the crude sample and washing of the bed in a fluidized mode and elusion in a fixed bed mode. A completely cl...

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Bibliographic Details
Published in:Biotechnology and bioengineering 1995-02, Vol.45 (3), p.205-211
Main Authors: Thommes, J. (Heinrich-Heine Universitat Dusseldorf, Julich, Germany.), Halfar, M, Lenz, S, Kula, M.R
Format: Article
Language:English
Subjects:
ANIMAL
Animal cells
ANIMALES
ANTICORPS MONOCLONAL
ANTICUERPOS MONOCLONALES
Biological and medical sciences
Biotechnology
Cell hybridization, cell fusion, hybridomas
CELLULE
CELULAS
Eukaryotic cell cultures
FERMENTACION
FERMENTATION
fluidized bed adsorption
Fundamental and applied biological sciences. Psychology
Methods. Procedures. Technologies
monoclonal antibodies
PURIFICACION
PURIFICATION
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Summary:To achieve the coarse purification of a monoclonal antibody from whole hybridoma fermentation broth a fluidized bed cation exchange process was used. The procedure consisted of application of the crude sample and washing of the bed in a fluidized mode and elusion in a fixed bed mode. A completely clarified eluate was obtained with purification factors between 4 and 8 and a concentration of the desired product (monoclonal antibody) by a factor of more than 3 was achieved. Thus, a combination of the three early steps of the downstream process clarification, concentration and coarse purification was possible. Two different materials were tested: a commercially available agarose-based matrix (Streamline-SP), and a self-derivatized material based on controlled-pore glass (Bioran). Initial experiments were performed to describe the fluidization of the glass material. Comparison with the agarose material showed several differences, the agarose matrix allowing liquid flow closer to plug flow than the glass material. Increased backmixing in the liquid phase was detected when fluidizing the glass adsorbent compared with the agarose-based matrix. Despite this fact, comparison of the two materials with respect to antibody binding and elusion demonstrated a similar performance
ISSN:0006-3592
1097-0290
DOI:10.1002/bit.260450304