Effective production of anti-tetanus toxoid and anti-HBsAg human monoclonal antibodies by serum-free culture of hybridomas

• Published in: Cytotechnology (Dordrecht) 1991, Vol.5 Suppl 2 (S2), p.S53-74
• Main Authors: Kitano, K, Ichimori, Y, Sawada, H, Iwasa, S, Sasai, S, Tsukamoto, K
• Format: Article
• Language: English
• Subjects: Animals; Antibodies, Monoclonal - biosynthesis; Antibodies, Monoclonal - immunology; Biotechnology; Biotechnology - methods; Cell Line; Culture Media; Hepatitis B Surface Antigens - immunology; Humans; Hybridomas - metabolism; Tetanus Toxoid - immunology
• ISSN: 0920-9069; 1573-0778
• DOI: 10.1007/BF00573880

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.