Solubilization of proteins in aqueous two-phase extraction through combinations of phase-formers and displacement agents

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• 作者：Christian Kress ; Gabriele Sadowski ; Christoph Brandenbusch ; ^{christoph.brandenbusch@bci.tu-dortmund.de}
• 关键词：ATPE ; aqueous two-phase extraction ; ATPS ; aqueous two-phase system ; CG-MALS ; composition gradient multi-angle light scattering ; DA ; displacement agent ; ePC-SAFT ; electrolyte Perturbed-Chain Statistical Associating Fluid Theory ; HSA ; human serum albumin ; IgG ; immunoglobulin G ; mAb ; monoclonal antibody ; PEG ; polyethylene glycol ; TLL ; tie line length ; TLS ; tie line slope ; UV ; ultra violet
• 刊名：European Journal of Pharmaceutics and Biopharmaceutics
• 出版年：2017
• 出版时间：March 2017
• 年：2017
• 卷：112
• 期：Complete
• 页码：38-44
• 全文大小：662 K
• 卷排序：112

文摘

The aqueous two-phase extraction (ATPE) of therapeutic proteins is a promising separation alternative to cost-intensive chromatography, still being the workhorse of nowadays downstream processing. As shown in many publications, using NaCl as displacement agent in salt-polymer ATPE allows for a selective purification of the target protein immunoglobulin G (IgG) from human serum albumin (HSA, represents the impurity). However a high yield of the target protein is only achievable as long as the protein is stabilized in solution and not precipitated. In this work the combined influence of NaCl and polyethylene glycol (Mw = 2000 g/mol) on the IgG-IgG interactions was determined using composition gradient multi-angle light scattering (CG-MALS) demonstrating that NaCl induces a solubilization of IgG in polyethylene glycol 2000 solution. Moreover it is shown that the displacement agent NaCl has a significant and beneficial influence on the IgG solubility in polyethylene glycol 2000-citrate aqueous two-phase system (ATPS) which can also be accessed by these advanced B22 measurements. By simultaneous consideration of IgG solubility data with results of the ATPS phase behavior (especially volume fraction of the respective phases) allows for the selection of process tailored ATPS including identification of the maximum protein feed concentration. Through this approach an ATPS optimization is accessible providing high yields and selectivity of the target protein (IgG).