Therapeutic antibodies have become one of the most effective therapeutics for human diseases such as cancer, inflammation and viral infection. The production of antibody-based drugs using microbial expression systems is more cost effective with ease of gene manipulation compared to mammalian expression systems. In our team, the antibody fragments (scFv and BsAb) were produced from methylotrophic yeast Pichia pastoris secretion expression system with the AOX1 as driven promoter. And the other on platform, Escherichia coli expression system, can effective product Fab. We could development and construct secretion expression cell line which such like E. coli and P. pastoris. High production cell line came from high through put screening. The optimal fermentation parameters were using 250 mL parallel mini-fermenters to define and testing. The 5L fermenter was using to confirm fermentation parameters which from mini-fermenter. In this study about P. pastoris, we found significant relationship from screening to 5L fermenter. One of the best scFv production yielded form deep well reach to1 mg/L, production form 50 ml shake flask stage reach to 5 mg/L, production form 250 ml shake flask stage reach to 20 mg/L and production form 250 ml mini-fermenter stage is over 500 mg/L, with amplification fold around 500 fold. E. coli had same trend but the expression yield could not as same as P. pastoris which only one tenth or one fifth. The largest scale fermenter for E. coli tills to 30 liters. We just not only scale up parameters from min-fermenter to 30L but also scale down 30L operation condition to 5L.

Engineering of microbial expression platform for antibodies fragments with scale-up process

253rd National Meeting (2017)

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