A SIMPLE AND INEXPENSIVE HIGH-DENSITY DIALYSIS TUBING CELL-CULTURE SYSTEM FOR THE IN-VITRO PRODUCTION OF MONOCLONAL-ANTIBODIES IN HIGH-CONCENTRATION

This paper describes the construction and application of a low-cost ro ller bottle-like culture appliance in which hybridoma cells can be cul tivated in high density in dialysis tubing. The appliance facilitates the simultaneous culture of up to four cell lines yielding 50 ml cultu re volume of each. Samples for follow-up analysis of the cultures can easily be taken when needed through sample ports. In order to obtain h igh cell densities (at least 10(7) cells/ml), high cell viability (at least 50%) and high antibody yield (at least 1.0 mg/ml) the bottle is rolled at a speed of 4-6 rpm and is gassed continuously by a micropump driven by rechargeable NiCd batteries fixed to the culture flask. Dep ending on the individual properties of the hybridoma lines tested, the cells may be cultured for 1-2 weeks, and cell densities of up to 30 x 10(6) cells/ml with viabilities of approximately 50% and monoclonal a ntibodies in concentrations of up to 2.8 mg/ml may be obtained. In the ir properties the monoclonal antibodies produced by this in vitro proc edure are indistinguishable from those prepared in the form of convent ional stationary culture supernatant or of ascitic fluid. Specific ant ibody content is within the same range as in ascitic fluid. Consequent ly, the monoclonal antibodies can be purified in one step, e.g., by io n exchange chromatography from the culture supernatant. Therefore. the newly developed culture device and the culture method described is a useful alternative to ascites production in live mice.