Continuous capture of recombinant antibodies by ZnCl2 precipitation without polyethylene glycol
Autor: | Peter Satzer, Daniel Komuczki, Alois Jungbauer, Gregory Dutra |
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Jazyk: | angličtina |
Rok vydání: | 2020 |
Předmět: |
Environmental Engineering
Chromatography Downstream processing purification Precipitation (chemistry) IgG Membrane fouling zinc Bioengineering Polyethylene glycol precipitation Cross-flow filtration Dilution Viscosity chemistry.chemical_compound downstream processing chemistry PEG ratio Research Articles Biotechnology Research Article |
Zdroj: | Engineering in Life Sciences |
ISSN: | 1618-2863 1618-0240 |
Popis: | The capture of recombinant antibodies from cell culture broth is the first critical step of downstream processing. We were able to develop a precipitation-based method for the capture and purification of monoclonal antibodies based on divalent cations, namely ZnCl2. Traditional precipitation processes have to deal with high dilution factors especially for resolubilization and higher viscosity due to the use of PEG as precipitation or co-precipitation agent. By the use of the crosslinking nature of divalent cations without the use of PEG, we kept viscosity from the supernatant and resolubilization dilution factors very low. This is especially beneficial for the solid–liquid separation for the harvest and wash of the precipitate in continuous mode. For this harvest and wash, we used tangential flow filtration that benefits a lot from low viscosity solutions, which minimizes the membrane fouling. With this precipitation based on ZnCl2, we were able to implement a very lean and efficient process. We demonstrated precipitation studies with three different antibodies, Adalimumab, Trastuzumab, and Denosumab, and a continuous capture case study using tangential flow filtration for precipitate recovery. In this study, we achieved yields of 70%. |
Databáze: | OpenAIRE |
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