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Purification of monoclonal antibodies from whole hybridoma fermentation broth by fluidized bed adsorption
Author(s) -
Thömmes Jörg,
Halfar Markus,
Lenz Suzanne,
Kula MariaRegina
Publication year - 1995
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260450304
Subject(s) - chromatography , agarose , elution , chemistry , fluidized bed , adsorption , fluidization , matrix (chemical analysis) , downstream processing , fermentation , monoclonal antibody , expanded bed adsorption , bioreactor , biochemistry , antibody , organic chemistry , immunology , biology
Abstract To achive 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 elution 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 achived. 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 (Stream‐line‐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 elution demonstrated a similar performance. © 1995 John Wiley & Sons, Inc.