title: Effective production of anti-tetanus toxoid and anti-HBsAg human monoclonal antibodies by serum-free culture of hybridomas.
abstract: 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 semicontinuous 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.
authors.affiliation: Research and Development Division, Takeda Chemical Industries, Ltd., Jusohonmachi 2-chome, Yodogawa-ku, 532, Osaka, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Kitano
authors.forename: Y
authors.initials: Y
authors.lastname: Ichimori
authors.forename: H
authors.initials: H
authors.lastname: Sawada
authors.forename: S
authors.initials: S
authors.lastname: Iwasa
authors.forename: S
authors.initials: S
authors.lastname: Sasai
authors.forename: K
authors.initials: K
authors.lastname: Tsukamoto