Learning from the mammalian expression system to develop a high titer- half antibody process in E. coli

While producing half-antibodies for bispecific antibody therapeutics in E. coli has its advantages over a mammalian expression system, it also comes with challenges associated with soluble expression and effective folding of these complex proteins. The merits and challenges of using a microbial host for half-antibody production will be discussed, with a focus on a chaperone overexpression strategy to achieve high titers in a short amount of time. To develop a high titer E. coli process, we looked to the mammalian expression system to better understand the mechanisms involved in antibody folding. By overexpressing the endogenous E. coli protein FkpA, we can facilitate similar folding mechanisms, suggesting that FkpA could be acting as an analog to both the mammalian chaperone BiP and ppiase CypB involved in antibody folding in CHO cells. Please click Additional Files below to see the full abstract