(210b) N/a

The controlled production of artificial antibodies for therapeutic treatments is a rapidly growing and profitable field. One primary method of antibody production is transient transfection in mammalian cells most often using polyethyleneimine (PEI) as the transfection agent and Human Embryonic Kidney (HEK) or Chinese Hamster Ovary (CHO) stable cell lines. Despite its importance, transfection is not well understood and leads to variable results based on many factors including medium used, transfection and incubation times, viable cell density (VCD) at inoculation, PEI concentration, and DNA concentration. The quantity and quality of antibody produced after transfection are of the upmost importance. In this experiment, a specific transfection protocol was optimized for antibody titer in HEK cells. Design of Experiments software was utilized to plan experiments to maximize antibody titer by varying the parameters of inoculation VCD, PEI concentration, and transfection time. Experiments were carried out in a six-chamber HexaScreen® mini-bioreactor with a control and scale-up performed with a 2 liter roller bottle system. Using the HexaScreen, 15 different trials were performed over the given parameters to conclude that the optimal combination was an inoculation VCD of 3 million cells/mL, a PEI concentration of 35 mg/L, and no transfection time, which would yield an antibody concentration over 40 mg/L. The roller bottle scale-up demonstrated inconsistency and scale-up problems that would need to be addressed with future experiments. While further experiments are necessary, this project demonstrated the feasibility of this system for better understanding and optimizing transient transfection and some of the relations between antibody titer and transfection parameters.