Selective generation of antigen-specific human hybridomas optimized for large scale growth in serum-free medium.
The direct propagation of newly formed human hybridomas in serum-free medium selects for hybrids with a metabolism best suited to growth in this environment. Under optimal culture conditions, this procedure results in the generation of antigen-specific human hybridomas comparable in frequency, stability, and antibody secretion rate to that obtained with murine hybridomas. After a transient phase of a few days in the appropriate selection medium supplemented with 1% serum, hybridomas grow in serum-free medium in stationary cultures with a cell doubling time of 15-25 h and an antibody production rate averaging 12 micrograms/10(6) cells/day. Clones propagated in bioreactors exhibited a cell doubling time of 29-35 h and an antibody secretion rate of 10-21 micrograms/10(6) cells/day.
Lang
A B
AB
Department of Immunology, Swiss Serum and Vaccine Institute, Berne.
Schürch
U
U
Zimmermann
F
F
Bruderer
U
U
D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology; D000918:Antibody Specificity; D001402:B-Lymphocytes; D002455:Cell Division; D002459:Cell Fusion; D002478:Cells, Cultured; D002999:Clone Cells; D016895:Culture Media, Serum-Free; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D007074:Immunoglobulin G / Q000096:biosynthesis; D007075:Immunoglobulin M / Q000096:biosynthesis; D007145:Immunoglobulin kappa-Chains / Q000096:biosynthesis; D012641:Selection, Genetic
D000911:Antibodies, Monoclonal; D016895:Culture Media, Serum-Free; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D007145:Immunoglobulin kappa-Chains