Temperature-dependent aqueous micellar two-phase systems (AMTPSs) have recently been gaining attention in the isolation of high-added-value biomolecules from their natural sources. Despite their sustainability, aqueous two-phase systems, and particularly AMTPSs, have not been extensively applied in the industry, which might be changed by applying process integration and continuous manufacturing. Here, we report for the first time on an integrated microfluidic platform for fast and low-material-consuming development of continuous protein purification using an AMTPS. A system comprised of a microchannel incubated at high temperature, enabling instantaneous triggering of a two-phase system formation, and a microsettler, allowing complete phase separation at the outlets, is reported here. The separation of phycobiliproteins and particularly the purification of R-phycoerythrin from the contaminant proteins present in the aqueous crude extract obtained from fresh cells of Gracilaria gracilis were thereby achieved. The results from the developed microfluidic system revealed that the fractionation performance was maintained while reducing the processing time more than 20-fold when compared with the conventional lab-scale batch process. Furthermore, the integration of a miniaturized ultrafiltration module resulted in the complete removal of the surfactant from the bottom phase containing R-phycoerythrin, as well as in nearly twofold target protein concentration. The process setup successfully exploits the benefits of process intensification along with the integration of various downstream processes. Further transfer to a meso-scale integrated system would make such a system appropriate for the separation and purification of biomolecules with high commercial interest.

中文翻译：

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胶束水两相系统用于R-藻红蛋白纯化的微流体平台的开发。

依赖温度的水性胶束两相系统（AMTPS）最近在从其天然来源中分离高附加值生物分子中引起了人们的注意。尽管具有可持续性，但水性两相系统，尤其是AMTPS，尚未在工业中得到广泛应用，可能会通过应用过程集成和连续制造而发生变化。在这里，我们首次在集成的微流控平台上报告了使用AMTPS进行连续蛋白纯化的快速，低耗材开发。本文报道了一个系统，该系统包括一个在高温下孵育的微通道，该模块可即时触发两相系统的形成；一个微沉降器，可在出口处实现完全的相分离。由此实现了Gracilaria gracilis。与常规实验室规模的分批工艺相比，开发的微流体系统的结果表明，在保持分馏性能的同时，将处理时间减少了20倍以上。此外，微型化超滤模块的整合导致表面活性剂从含有R-藻红蛋白的底部相中完全去除，并使目标蛋白质浓度接近两倍。流程设置成功地利用了流程强化以及集成各种下游流程的好处。进一步转移到中等规模的集成系统将使这种系统适合于具有高商业兴趣的生物分子的分离和纯化。