Hybridoma cell growth and anti-neuroblastoma monoclonal antibody production in spinner flasks using a protein-free medium with microcarriers.
The main disadvantages of foetal calf serum as the world-wide common serum supplement for cell growth are its content of various proteins of variable concentrations between batches as well as its high cost. The use of serum-free and protein-free media is gradually becoming one of the goals of cell culture especially for standardizing culture conditions or for simple purification of cell products like monoclonal antibodies. The mouse hybridoma cells 14/2/1 were cultivated either in protein-free UltraDOMA medium or in serum-containing RPMI medium with and without microcarriers to generate high quantities of monoclonal antibodies against neuroblastoma tumour cells. Cell growth rate, IgG production, viability, glucose and lactate concentrations, attachment rate and doubling time have been used as investigation criteria. Modifications of culture procedures (static or stirred), inoculum density, and microcarrier concentration caused an improvement of monoclonal antibody production. The kinetics of antibody synthesis was best in spinner culture with 2 ml of microcarriers in protein-free medium. These results of short-term microcarrier culture in stirred spinner flasks indicate that IgG yields in protein-free medium 2.5-fold higher to those in serum-supplemented medium can be achieved.
Voigt
A
A
Children's Hospital, Department of Hematology, Oncology and Immunology, University of Jena, Germany. avoi@kikli1.med.uni-jena.de
Zintl
F
F
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D018929:Cell Culture Techniques; D003470:Culture Media; D006825:Hybridomas / Q000166:cytology* / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D051379:Mice; D009447:Neuroblastoma / Q000276:immunology*
D000911:Antibodies, Monoclonal; D003470:Culture Media; D007074:Immunoglobulin G