Effective production of anti-tetanus toxoid and anti-HBsAg human monoclonal antibodies by serum-free culture of hybridomas.
Two hybridoma systems, mouse.human-human (m.h-h) heterohybridoma and human-human (h-h) hybridoma, have been established, and hybridomas secreting anti-tetanus toxoid and anti-HBsAg human monoclonal antibodies (MoAbs), both having a neutralizing activity have been obtained. Cell-line improvement was shown to be an efficient method for improving the productivity in a cell culture process. Two kinds of serum-free media, GFS (a serum substitute)-containing media and polyethylene glycol (PEG)-containing media, have been established to produce human MoAbs. m.h-h Heterohybridomas could be cultivated for a long period by perfusion culture in an agitation vessel, but h-h hybridomas could not. We found that h-h hybridomas show growth-associated antibody production kinetics and established two kinds of long-term cultivation systems: continuous perfusion culture and semi-continuous immobilized perfusion culture. We also scaled up batch culture and short-term perfusion culture to 200-L and 50-L fermentors, respectively. Processes for large-scale purification from the culture supernatants of both GFS- and PEG-containing serum-free media have also been developed.
Research and Development Division, Takeda Chemical Industries, Ltd., Osaka, Japan.
K
K
Kitano
Y
Y
Ichimori
H
H
Sawada
S
S
Iwasa
S
S
Sasai
K
K
Tsukamoto
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D001709:Biotechnology / Q000379:methods; D002460:Cell Line; D003470:Culture Media; D006514:Hepatitis B Surface Antigens / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000378:metabolism*; D013745:Tetanus Toxoid / Q000276:immunology*
D000911:Antibodies, Monoclonal; D003470:Culture Media; D006514:Hepatitis B Surface Antigens; D013745:Tetanus Toxoid