Electrical stimulation of hybridoma cells producing monoclonal antibody to cAMP.
Electrical stimulation was applied to hybridoma cells in order to activate metabolic activities and increase the monoclonal antibody production. Hybridoma cells that produce monoclonal antibody to adenosine 3':5'-cyclic monophosphate were placed on a transparent glass electrode immersed in medium and subjected to electric pulses (pulse shape, alternating rectangular; field strength, 4 X 10(3) V X m-1; frequency, 5 kHz; pulse mode, 0.5 min application and 4.5 min pause). After 48 h of incubation, the concentration of lactic acid in the medium reached 8.4 mM, approx. 30% higher than that obtained without electric stimulation. Similarly, cell growth rate was promoted by the electric stimulation, reaching a maximum stimulation after 40 h. When the hybridoma was cultured for 48 h with electrical stimulation, the antibody concentration in the medium reached 22.3 microgram X ml-1, approx. 10% higher than the control, with a concomitant 16% increase in cell concentration. Longer periods of electric pulse application, however, caused an inhibitory effect on the hybridoma growth. The most probable cause of the inhibition are reactive oxygen species such as superoxide and hydrogen peroxide, which are inevitably generated by electrolysis. The presence of superoxide dismutase (EC 1.15.1.1) reduced the inhibitory effects. In conclusion, metabolic activities including monoclonal antibody production were activated by the electrical stimulation.
D000818:Animals; D000911:Antibodies, Monoclonal; D002374:Catalase / Q000378:metabolism; D000242:Cyclic AMP / Q000276:immunology*; D004558:Electric Stimulation; D004563:Electrochemistry; D006825:Hybridomas / Q000378:metabolism*; D007773:Lactates / Q000096:biosynthesis; D019344:Lactic Acid; D051379:Mice; D008807:Mice, Inbred BALB C; D013482:Superoxide Dismutase / Q000378:metabolism
D000911:Antibodies, Monoclonal; D007773:Lactates; D019344:Lactic Acid; D000242:Cyclic AMP; D002374:Catalase; D013482:Superoxide Dismutase