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Precipitation of complex antibody solutions: influence of contaminant composition and cell culture medium on the precipitation behavior

Großhans, Steffen; Suhm, Susanna; Hubbuch, Jürgen

Abstract:
Preparative protein precipitation is known as a cost-efficient and easy-to-use alternative to chromatographic purification steps. This said, at the moment, there is no process for monoclonal antibodies (mAb) on the market, although especially polyethylene glycol-induced precipitation has shown great potential. One reason might be the highly complex behavior of each component of a crude feedstock during the precipitation process. For different investigated mAbs, significant variations in the host cell protein (HCP) reduction are observed. In contrast to the precipitation behavior of single components, the interactions and interplay in a complex feedstock are not fully understood yet. This work discusses the influence of contaminants on the precipitation behavior of two different mAbs, an IgG1, and an IgG2. By spiking the mAbs with mock solution, a complex feedstock could successfully be mimicked. Spiking contaminants influenced the yield and purity of the mAbs after the precipitation step, compared to the precipitation behavior of the single components. The mixture showed a decrease in the contaminant and mAb solubility. By re-buffering the mock solution prior to spiking, special salts, small molecules like amino acids, vitamins, or sugars could be depleted while larger ones like HCP or DNA were still present. ... mehr

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Verlagsausgabe §
DOI: 10.5445/IR/1000094447
Veröffentlicht am 17.06.2019
Originalveröffentlichung
DOI: 10.1007/s00449-019-02103-y
Scopus
Zitationen: 1
Web of Science
Zitationen: 1
Cover der Publikation
Zugehörige Institution(en) am KIT Karlsruher Institut für Technologie (KIT)
Publikationstyp Zeitschriftenaufsatz
Publikationsjahr 2019
Sprache Englisch
Identifikator ISSN: 0178-515X, 1432-0797, 1615-7591, 1615-7605
KITopen-ID: 1000094447
Erschienen in Bioprocess and biosystems engineering
Band 42
Heft 6
Seiten 1039-1051
Projektinformation nextBioPharmDSP (EU, H2020, 635557)
Nachgewiesen in Web of Science
Scopus
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