Despite the benefits of aqueous two-phase extraction (ATPE) for purification of high-value biomolecules, phase-formers that are used to generate the aqueous two-phase system (ATPS) often reduce the stability and solubility of the biomolecule. Within this work, we thus evaluated the use of suitable excipients to increase the biomolecule stability and solubility during ATPE, whilst not negatively affecting yield and selectivity. Applying this new physically sound approach enabled a rapid selection of L-arginine as suitable excipient for the ATPE of Immunoglobulin G. It was demonstrated that both the (conformational and colloidal) stability and solubility of Immunoglobulin G in the ATPS were increased, thus maximizing yield, minimizing precipitation, and improving the extraction performance of the ATPS. The applicability and transferability of the approach was confirmed by optimizing the ATPE of an industrial relevant amine dehydrogenase. The optimized ATPS enabled an extraction yield of 79.17 wt% (impurities < 7 wt%) directly from crude extract. Furthermore, using the excipients broadened the ATPE process window, thus simplifying process handling during ATPE and further downstream unit operations. Prospectively, the use of excipients can support the establishment of ATPE as downstream technology.

尽管水相两相萃取 (ATPE) 对纯化高价值生物分子有好处，但用于生成水相两相系统 (ATPS) 的相形成剂通常会降低生物分子的稳定性和溶解度。在这项工作中，我们因此评估了使用合适的赋形剂来增加 ATPE 期间生物分子稳定性和溶解性，同时不会对产量和选择性产生负面影响。应用这种新的物理上合理的方法可以快速选择 L-精氨酸作为免疫球蛋白 G 的 ATPE 的合适赋形剂。 结果表明，免疫球蛋白 G 在 ATPS 中的（构象和胶体）稳定性和溶解度都增加了，从而最大限度地提高了产量，最大限度地减少沉淀，并提高 ATPS 的提取性能。通过优化工业相关胺脱氢酶的 ATPE，证实了该方法的适用性和可转移性。优化后的 ATPS 能够直接从粗提物中提取 79.17 wt%（杂质 < 7 wt%）的提取率。此外，使用赋形剂拓宽了 ATPE 工艺窗口，从而简化了 ATPE 和进一步下游单元操作期间的工艺处理。前瞻性地，赋形剂的使用可以支持ATPE作为下游技术的建立。从而简化 ATPE 和进一步下游单元操作期间的工艺处理。前瞻性地，赋形剂的使用可以支持ATPE作为下游技术的建立。从而简化 ATPE 和进一步下游单元操作期间的工艺处理。前瞻性地，赋形剂的使用可以支持ATPE作为下游技术的建立。