Monoclonal antibodies (mAbs) offer many advantages for the treatment of chronic and inflammatory diseases over traditional small molecule therapeutics. Indeed, their proven value as therapeutics sparked major interest from pharmaceutical companies for increased development and marketing with the global market for mAbs predicted to reach $1.2 trillion USD by 2020. While unrivaled specificity and a dearth of adverse effects shine as some of the most desirable characteristics, the cost of production, stability issues from manufacturing to delivery, and requisite high concentrations for physiological efficiency are issues which still need consideration.<br/><br/>mAbs share many common structural features, so understanding how each part contributes to structural characteristics of the entire molecule may unveil ways to design better therapeutics. The Fc portion of a mAb enables purification via Protein A chromatography and controls antibody effector functions (e.g. antibody dependent cell-mediated cytotoxicity, complement dependent cytotoxicity) depending on glycoform presence and identity. Characterizing both aglycosylated and glycosylated Fc and their contributions to antibody stability requires adequate recombinant protein yield from mammalian cell culture.<br/><br/>Mammalian cell cultures (e.g. CHO and HEK) used for protein production are either stable or transiently transfected, and lipofection and polyfection represent the most common methods of performing the transfection. Here, we determine head-to-head comparison of the yields of recombinant Fc protein from transient/stable and lipid/polymer mediated transfections in 30 mL scale cultures of HEK Freestyle 293 and ATCC HEK293 cells. Heterogeneous populations of stably transfected HEK cells under Zeocin™ antibiotic selection produced 5-10 mg/L yields, but further passages of these cells reduced overall production. The proprietary cationic lipid formulations resulted in the highest overall Fc amounts (5-15 mg/L) in transiently transfected cells, but PEI mediated polyfection (yielding &gt;2 mg/L) could offer greater overall cost efficiency and scalability under similar conditions.

Comparison of transfection methods on yield of recombinant human IgG1 Fc

255th National Meeting (2018)

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