title: Ethanol precipitation for purification of recombinant antibodies.
abstract: Currently, the golden standard for the purification of recombinant humanized antibodies (rhAbs) from CHO cell culture is protein A chromatography. However, due to increasing rhAbs titers alternative methods have come into focus. A new strategy for purification of recombinant human antibodies from CHO cell culture supernatant based on cold ethanol precipitation (CEP) and CaCl2 precipitation has been developed. This method is based on the cold ethanol precipitation, the process used for purification of antibodies and other components from blood plasma. We proof the applicability of the developed process for four different antibodies resulting in similar yield and purity as a protein A chromatography based process. This process can be further improved using an anion-exchange chromatography in flowthrough mode e.g. a monolith as last step so that residual host cell protein is reduced to a minimum. Beside the ethanol based process, our data also suggest that ethanol could be replaced with methanol or isopropanol. The process is suited for continuous operation.
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, A-1190 Vienna, Austria.
authors.forename: Anne
authors.initials: A
authors.lastname: Tscheliessnig
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, A-1190 Vienna, Austria.
authors.forename: Peter
authors.initials: P
authors.lastname: Satzer
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, A-1190 Vienna, Austria; Austrian Centre of Industrial Biotechnology, Vienna, Muthgasse 18, 1190 Vienna, Austria.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Hammerschmidt
authors.affiliation: Novartis Pharma AG, CH-4002 Basel, Switzerland.
authors.forename: Henk
authors.initials: H
authors.lastname: Schulz
authors.affiliation: Novartis Pharma AG, CH-4002 Basel, Switzerland.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Helk
authors.affiliation: Department of Biotechnology, University of Natural Resources and Life Sciences, Vienna, A-1190 Vienna, Austria; Austrian Centre of Industrial Biotechnology, Vienna, Muthgasse 18, 1190 Vienna, Austria. Electronic address: alois.jungbauer@boku.ac.at.
authors.forename: Alois
authors.initials: A
authors.lastname: Jungbauer
mesh_terms: D000818:Animals; D000906:Antibodies / Q000302:isolation & purification*; D016466:CHO Cells; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D006224:Cricetinae; D003412:Cricetulus; D004797:Enzyme-Linked Immunosorbent Assay; D000431:Ethanol / Q000737:chemistry*; D011994:Recombinant Proteins / Q000302:isolation & purification; D013205:Staphylococcal Protein A / Q000737:chemistry
chemical_list: D000906:Antibodies; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; D000431:Ethanol
keywords: CHO; Cold ethanol precipitation; Monolith; Organic solvent; Recombinant antibodies