Abstract：
Proteins in ionic liquids (ILs) and deep eutectic solvents (DESs) have gained significant attention due to their potential applications in various fields, including biocatalysis, bioseparation, biomolecular delivery, and structural biology. Scattering approaches including dynamic light scattering (DLS) and small-angle X-ray and neutron scattering (SAXS and SANS) have been used to understand the solution behavior of proteins at the nanoscale and microscale. This review provides a thorough exploration of the application of these scattering techniques to elucidate protein properties in ILs and DESs. Specifically, the review begins with the theoretical foundations of the relevant scattering approaches and describes the essential solvent properties of ILs and DESs linked to scattering such as refractive index, scattering length density, ion-pairs, liquid nanostructure, solvent aggregation, and specific ion effects. Next, a detailed introduction is provided on protein properties such as type, concentration, size, flexibility and structure as observed through scattering methodologies. This is followed by a review of the literature on the use of scattering for proteins in ILs and DESs. It is highlighted that enhanced data analysis and modeling tools are necessary for assessing protein flexibility and structure, and for understanding protein hydration, aggregation and specific ion effects. It is also noted that complementary approaches are recommended for comprehensively understanding the behavior of proteins in solution due to the complex interplay of factors, including ion-binding, dynamic hydration, intermolecular interactions, and specific ion effects. Finally, the challenges and potential research directions for this field are proposed, including experimental design, data analysis approaches, and supporting methods to obtain fundamental understandings of complex protein behavior and protein systems in solution. We envisage that this review will support further studies of protein interface science, and in particular studies on solvent and ion effects on proteins.
摘要：
离子液体(ILs)和低共熔溶剂(DESs)中的蛋白质因其在各个领域的潜在应用而受到广泛关注。包括生物催化,生物分离,生物分子传递，结构生物学。包括动态光散射（DLS）和小角度X射线和中子散射（SAXS和SANS）的散射方法已用于了解蛋白质在纳米级和微米级的溶液行为。这篇综述提供了这些散射技术在阐明IL和DES中蛋白质特性方面的应用。具体来说,该综述从相关散射方法的理论基础开始，并描述了与散射有关的IL和DES的基本溶剂性质，例如折射率，散射长度密度，离子对，液体纳米结构，溶剂聚集，和特定的离子效应。接下来,详细介绍了蛋白质的性质，如类型，浓度,尺寸,通过散射方法观察到的灵活性和结构。随后是有关在IL和DES中使用蛋白质散射的文献的综述。强调增强的数据分析和建模工具对于评估蛋白质的灵活性和结构是必要的。为了理解蛋白质的水合作用，聚集和特定离子效应。还应注意，由于因素的复杂相互作用，建议采用补充方法来全面了解溶液中蛋白质的行为，包括离子结合，动态水化,分子间相互作用，和特定的离子效应。最后,提出了该领域面临的挑战和潜在的研究方向，包括实验设计，数据分析方法，和支持方法，以获得对溶液中复杂蛋白质行为和蛋白质系统的基本理解。我们设想这篇综述将支持蛋白质界面科学的进一步研究，特别是关于溶剂和离子对蛋白质影响的研究。
