An alternative downstream process based on aqueous two-phase extraction for the purification of monoclonal antibodies

•New alternative downstream process based on ATPS without protein A chromatography.•A holistic development as well as optimization of the individual unit operations is demonstrated.•74 % yield of mAb with simultaneous removal of DNA (99.97 %) and HCP (97.11 %).•mAb glycosylation pattern remained uni...

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Bibliographic Details
Published inBiochemical engineering journal Vol. 161; p. 107703
Main Authors Kruse, Thomas, Kampmann, Markus, Rüddel, Ines, Greller, Gerhard
Format Journal Article
LanguageEnglish
Published Elsevier B.V 15.09.2020
Subjects
Aqueous two-phase extraction
ATPS
Chromatography
Diafiltration
Downstream
Monoclonal antibody
ATPS
Monoclonal antibody
Diafiltration
Aqueous two-phase extraction
Downstream
Chromatography
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Summary:•New alternative downstream process based on ATPS without protein A chromatography.•A holistic development as well as optimization of the individual unit operations is demonstrated.•74 % yield of mAb with simultaneous removal of DNA (99.97 %) and HCP (97.11 %).•mAb glycosylation pattern remained unimpaired after ATPE and throughout the alternative DSP (stable product quality). An alternative downstream process for the purification of a monoclonal antibody (mAb) based on aqueous two-phase extraction as clarification, capture and primary purification step was developed. For further purification unit operations which are commonly used for mAb platform processes were utilized. A diafiltration approach was used to combine virus inactivation and removal of phase forming components as well as low molecular weight impurities in one step followed by cation and anion exchange chromatography. Starting from cell containing cultivation broth, an overall mAb yield of 74 % was achieved within an application study. The process was optimized regarding mAb yield and the clearance of process related impurities like deoxyribonucleic acid as well as host cell proteins, which were removed to approximately 60 and 6000 ppm respectively. Critical product quality attributes, regarding glycosylation patterns, were also examined and remained unimpaired after the aqueous two-phase extraction. The alternative downstream process presented in this study offers great potential to improve mAb manufacturing.
ISSN:1369-703X
DOI:10.1016/j.bej.2020.107703