As previously reported, fluoroalcohols can induce coacervation in aqueous solutions of amphiphilic compounds with subsequent formation of two-phase systems, where one phase is enriched in amphiphile and fluoroalcohol and the other is primarily an aqueous – rich phase. This study focuses on the use of simple coacervates made of a single component amphiphile induced by a fluoroalcohol for extraction and enrichment of proteins. 1,1,1,3,3,3-Hexafluoroisopropanol (HFIP) and 2,2,2-trifluoroethanol (TFE) were used to induce coacervation in the aqueous solutions of a cationic amphiphile, cetyltrimethylammonium bromide (CTAB) or tetra-n-butylammonium bromide (TBAB). Cationic amphiphiles (CTAB, TBAB) formed two-phase coacervate systems in a basic pH and/or sufficient ionic strength depending on the strength of coacervator (HFIP or TFE). The phase diagrams for TBAB paired with HFIP or TFE coacervates were created. By increasing the concentration of coacervator (HFIP or TFE) at a constant surfactant concentration, transition from a single liquid phase to a two or multiple phase mixture, and then eventually to a single liquid phase was observed. TBAB/HFIP mixture without additives showed a unique three-phase system before transitioning to a two-phase system upon increasing HFIP concentration. However, salt addition eliminated this three-phase region and expanded the region of two-phase formation. Select two-phase systems composed of TBAB and a perfluoroalcohol (HFIP or TFE) were utilized to extract model proteins of ranging hydrophobicity. All coacervate phases extracted bacteriorhodopsin, a membrane protein, and gramicidin, a very hydrophobic polypeptide ion channel. The most hydrophilic protein in the mixture, ribonuclease A, remained in aqueous phases. The coacervates formed from TBAB/TFE/200 mM NaCl mixture and TBAB/HFIP mixture exhibited the most selectivity in extracting proteins of high hydrophobicity. The partition coefficient (P) for each protein was calculated using the ratio of the protein concentration in the coacervate to that in the aqueous-rich phases. TBAB (50 mM)/HFIP (8%, v/v) coacervate showed remarkable selectivity and a high partition coefficient (>100) for both bacteriorhodopsin and gramicidin. Thus, this system may potentially be beneficial for facile fractionation of hydrophobic and membrane proteins in proteomics applications.

如先前报道，氟代醇可在两亲化合物的水溶液中引起凝聚，随后形成两相系统，其中一相富含两亲和氟代醇，而另一相则主要是富含水的相。这项研究的重点是将由氟代醇诱导的单组分两亲物制成的简单凝聚层用于蛋白质的提取和富集。1,1,1,3,3,3-六氟异丙醇（HFIP）和2,2,2-三氟乙醇（TFE）用于在阳离子两亲物，十六烷基三甲基溴化铵（CTAB）或四正丁基萘水溶液中诱导凝聚-丁基溴化铵（TBAB）。阳离子两亲物（CTAB，TBAB）在碱性pH和/或足够的离子强度（取决于凝聚器（HFIP或TFE）的强度）下形成了两相凝聚系统。创建了TBAB与HFIP或TFE凝聚层配对的相图。通过在恒定的表面活性剂浓度下增加凝聚器（HFIP或TFE）的浓度，观察到从单一液相过渡到两相或多相混合物，然后最终过渡到单一液相。不含添加剂的TBAB / HFIP混合物显示出独特的三相体系，然后随着HFIP浓度的增加转变为两相体系。但是，添加盐消除了该三相区域，并扩大了两相形成区域。选用由TBAB和全氟醇（HFIP或TFE）组成的两相系统来提取具有一定疏水性的模型蛋白。所有凝聚相均提取细菌视紫红质（一种膜蛋白）和短杆菌肽（一种非常疏水的多肽离子通道）。混合物中亲水性最高的蛋白质，核糖核酸酶A，保留在水相中。由TBAB / TFE / 200 mM NaCl混合物和TBAB / HFIP混合物形成的凝聚层在提取高疏水性蛋白质中表现出最大的选择性。使用凝聚层中的蛋白质浓度与富水相中的蛋白质浓度之比，计算出每种蛋白质的分配系数（P）。TBAB（50 mM）/ HFIP（8％，v / v）凝聚层对细菌视紫红质和短杆菌肽均显示出显着的选择性和高分配系数（> 100）。因此，