Studies on monoclonal antibody production by a hybridoma cell line (C1E3) immobilised in a fixed bed, porosphere culture system.
The aim of this study was to investigate the potential of fixed beds of macroporous glass spheres as a production process for animal cell products. The growth, metabolism and monoclonal antibody expression of a mouse-mouse hybridoma cell line was investigated in order to both test the potential of and to optimise the system. After the initial growth phase, the culture went into a steady-state phase brought on by glutamine limitation. An event occurred after 120-160 h of steady-state operation which destabilised the culture, causing a decline in productivity, after which the culture recovered. This event was analysed in detail to determine its cause, and whether a major switch in metabolic function had occurred. The parameter which correlated most closely to antibody production rate was oxygen, but as this was kept constant in the void medium of the bed it has to be concluded that oxygen diffusion into the spheres was the regulatory factor. A comparison of the fixed bed and a flask culture identified interesting differences in glucose metabolism between the two systems. The data gave strong indications as to how the productivity of the fixed bed system can be further improved. This includes optimisation of the glutamine concentration and modifying the porous structure of the spheres to improve diffusion characteristics.
Racher
A J
AJ
Division of Biologics, PHLS Centre for Applied Microbiology and Research, Porton Down, Salisbury, U.K.
Looby
D
D
Griffiths
J B
JB
D000641:Ammonia / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D002452:Cell Count; D002455:Cell Division; D002460:Cell Line; D002470:Cell Survival; D003584:Cytological Techniques; D005898:Glass; D005947:Glucose / Q000378:metabolism; D005973:Glutamine / Q000378:metabolism; D006825:Hybridomas / Q000166:cytology / Q000276:immunology* / Q000378:metabolism; D007700:Kinetics; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D007773:Lactates / Q000378:metabolism; D019344:Lactic Acid; D051379:Mice; D010100:Oxygen / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D007773:Lactates; D005973:Glutamine; D019344:Lactic Acid; D000641:Ammonia; D007770:L-Lactate Dehydrogenase; D005947:Glucose; D010100:Oxygen