Enhanced IgG production in eRDF media with and without serum. A comparative study.
The performance of three basal media RPMI, DMEM/F12 (DF) and eRDF (enhanced RDF, RPMI:DMEM:F12 in 2:1:1) were evaluated in cultures with and without serum with respect to cell proliferation, metabolism and monoclonal antibody (Mab) productivity. Based on the ease of adaptation, growth rate, maximum cell density and Mab production, the media were ranked as follows: eRDF > DF > RPMI. This was true for serum-free (SF) and serum supplemented (SS) media in static and shaker cultures. Growth performances in static and shaker cultures were consistently 20-50% lower in all three SF media compared to the corresponding SS conditions. Antibody titres in DF/SF and RPMI/SF cultures, irrespective of the culture condition, were generally similar or slightly lower than their SS counterparts. However, eRDF/SF medium yielded a much higher Mab titre (193 mg l-1) compared to eRDF/SS medium (145 mg l-1). This was also six times higher than the lowest titre of 30 mg l-1 in RPMI/SF medium. Hybridomas in eRDF/SF were further adapted to media without bovine serum albumin (eRDF/SF-BSA). Maximum cell densities in these cultures improved with scale up, from 1.1 x 10(6) ml-1 in static, to 1.9 x 10(6) ml-1 in shaker flasks, to 2.5 x 10(6) ml-1 in bioreactors. However, Ig levels remained between 100-130 mg l-1 which were much lower than in eRDF/SF medium. Thus BSA appears to be necessary for Ig production. The manufacturing cost (excluding purification) of Ig using eRDF was calculated to be between 17-50% of the price of the other two media and therefore this is regarded as the best medium for Ig production.
Chua
F
F
Chemical Engineering Department, National University of Singapore.
Oh
S K
SK
Yap
M
M
Teo
W K
WK
D000222:Adaptation, Physiological; D000818:Animals; D002455:Cell Division; D003470:Culture Media / Q000191:economics; D016895:Culture Media, Serum-Free / Q000191:economics; D046508:Culture Techniques / Q000191:economics / Q000379:methods*; D005947:Glucose / Q000378:metabolism; D006825:Hybridomas; D007074:Immunoglobulin G / Q000096:biosynthesis*; D051379:Mice
D003470:Culture Media; D016895:Culture Media, Serum-Free; D007074:Immunoglobulin G; D005947:Glucose