Feasibility studies of large scale production of human anti-tetanus toxoid monoclonal antibodies.
The feasibility of large scale production of human anti-tetanus toxoid monoclonal antibody for therapeutic use was evaluated using a human heterohybridoma. The effects of duration of subculture, transition from static to agitated culture conditions and the level of serum concentration were studied. The level of antibody secreted by the clone decreased with increasing length of subculture and decreasing serum concentration. The clone exhibited heterogeneity in expression of surface IgG after 2 or 7 weeks of subculture in static culture conditions irrespective of the serum concentration. However, a prolonged duration of subculture (9 weeks) in 3% serum medium had an effect on the expression of surface IgG both in static and agitated culture conditions. With respect to total (surface and intracellular) IgG, two distinct cell populations were observed. On long term subculture (9 weeks) in low serum medium (3% FCS), there was a decrease in the population which was the high synthesizer. In addition, when these cells were cultivated in agitated spinner flasks, a defect in secretion of antibodies was observed. Thus a general fall in the amount of antibody in the supernatant of agitated cultures was due to decrease in antibody synthesis as well as the defect in secretion of antibodies.
Martial-Gros
A
A
National Institute of Immunology, Aruna Asaf Ali Marg, New-Delhi, India.
Sarin
K K
KK
Mukhopadhyay
A
A
Ghosh
S
S
D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000627:therapeutic use; D018929:Cell Culture Techniques / Q000379:methods*; D003470:Culture Media / Q000737:chemistry; D004797:Enzyme-Linked Immunosorbent Assay; D005240:Feasibility Studies; D005434:Flow Cytometry; D006801:Humans; D006825:Hybridomas / Q000276:immunology* / Q000502:physiology; D013678:Technology, Pharmaceutical; D013745:Tetanus Toxoid / Q000276:immunology*