Novel Displacement Agents for Aqueous 2-Phase Extraction Can Be Estimated Based on Hybrid Shortcut Calculations.
Novel Displacement Agents for Aqueous 2-Phase Extraction Can Be Estimated Based on Hybrid Shortcut Calculations.
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可以根据混合捷径计算来估算用于水相两相萃取的新型驱替剂
DOI：
10.1016/j.xphs.2016.06.006
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发表时间：
2016
影响因子：
3.8
通讯作者：
Brandenbusch
中科院分区：
文献类型：
--
作者：
Sadowski;Brandenbusch
治疗性蛋白质的纯化是一项具有挑战性的任务，迫切需要优化。除了其他技术外，蛋白质的双水相萃取(ATPE)已被证明是一种有前途的替代成本密集的最先进的蛋白质层析纯化方法。最有可能的是，为了实现选择性提取，必须使用置换试剂来影响蛋白质的分离，以将目标蛋白质与杂质分离。在这项工作中，提出了一种新的驱替剂(溴化锂[LiBr])，它允许在柠檬酸-聚乙二醇ATPS中选择性地将目标蛋白IgG与人血清白蛋白(代表杂质)分离。为了表征溴化锂对免疫球蛋白的置换适合性，考察了溴化锂与形成相剂对双水相体系(ATPS)及分配的影响。利用组成梯度多角度光散射测量得到的渗透维里系数(B22和B23)，估算了LiBr2种蛋白质的沉淀效应和蛋白质分配系数。根据B22评估了溴化锂对这两种蛋白质的稳定作用，并在柠檬酸盐-聚乙二醇ATPS中进行了实验验证。我们的方法有助于在治疗性蛋白质的下游加工开发中有效地实施ATPE。
The purification of therapeutic proteins is a challenging task with immediate need for optimization. Besides other techniques, aqueous 2-phase extraction (ATPE) of proteins has been shown to be a promising alternative to cost-intensive state-of-the-art chromatographic protein purification. Most likely, to enable a selective extraction, protein partitioning has to be influenced using a displacement agent to isolate the target protein from the impurities. In this work, a new displacement agent (lithium bromide [LiBr]) allowing for the selective separation of the target protein IgG from human serum albumin (represents the impurity) within a citrate–polyethylene glycol (PEG) ATPS is presented. In order to characterize the displacement suitability of LiBr on IgG, the mutual influence of LiBr and the phase formers on the aqueous 2-phase system (ATPS) and partitioning is investigated. Using osmotic virial coefficients (B22andB23) accessible by composition gradient multiangle light-scattering measurements, the precipitating effect of LiBr on both proteins and an estimation of both protein partition coefficients is estimated. The stabilizing effect of LiBr on both proteins was estimated based onB22and experimentally validated within the citrate–PEG ATPS. Our approach contributes to an efficient implementation of ATPE within the downstream processing development of therapeutic proteins.
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影响因子：
2.6
作者：
T. Reschke;Christoph Brandenbusch;G. Sadowski
通讯作者：
G. Sadowski
使用第二渗透维里系数对蛋白质溶解度进行建模和预测
影响因子：
2.6
作者：
Marcel Herhut;Christoph Brandenbusch;G. Sadowski
通讯作者：
G. Sadowski
影响因子：
4.7
作者：
Rosa, Paula A. J.;Azevedo, Ana M.;Aires-Barros, M. Raquel
通讯作者：
Aires-Barros, M. Raquel
影响因子：
4.1
作者：
Christian Kress;G. Sadowski;Christoph Brandenbusch
通讯作者：
Christian Kress;G. Sadowski;Christoph Brandenbusch
DOI：
10.1021/je401002w
发表时间：
2014-03-01
期刊：
JOURNAL OF CHEMICAL AND ENGINEERING DATA
影响因子：
--
作者：
Glyk, Anna;Scheper, Thomas;Beutel, Sascha
通讯作者：
Beutel, Sascha