EAGER: Therapeutic Protein Separations via Surface Isoelectric Focusing (sIEF)

这一年，EAGER将探索一种新的和潜在的变革性方法，用于小体积蛋白质分离，称为表面等电聚焦（sIEF）。sIEF通过比以前的IEF技术小100倍的操作，提供了更简单、更便宜、更快速和更敏感的分析潜力。这项研究将扩展等电聚焦的知识，以非常高的分辨率分离和鉴定两性分子，例如蛋白质，这对涉及蛋白质异常的蛋白质病变的医学诊断和治疗性蛋白质的药物筛选至关重要。大规模平板凝胶的IEF是资源、劳动力和时间密集型的，在临床上被更小体积、基因组和亲和力的方法所取代。治疗蛋白的糖工程已被证明可以改善分子稳定性，调节物理化学和药理学性质，改善药代动力学，具有更好的吸收，更长的循环时间和更低的清除率。这项工作可以产生必要的知识，导致sIEF工具的开始，可以与治疗蛋白库无缝连接。结果将是快速筛选和有价值的电荷和等电点信息，以帮助开发治疗蛋白。

1548107Michigan Technological UniversityMinerickThis one year EAGER will explore a new and potentially transformative approach to small volume protein separations called surface isoelectric focusing (sIEF). sIEF offers the potential for simpler, cheaper, quicker, and sensitive analysis by operating 100 times smaller than previous IEF techniques. This research will expand knowledge in isoelectric focusing to separate and identify amphoteric molecules, e.g., proteins, at very high resolution, which is fundamental to medical diagnostics of proteopathy diseases involving protein abnormalities and pharmaceutical screening of therapeutic proteins. IEF in larger scale slab gels is resource, labor, and time intensive being replaced clinically by smaller volume, genomic and affinity approaches. Glycoengineering of therapeutic proteins has been shown to improve molecular stability, regulate physicochemical and pharmacological properties, and improve pharmacokinetics with better absorption, longer circulation times, and decreased clearance rates. This work could produce the knowledge necessary to lead to the beginning of an sIEF tool which could interface seamlessly with therapeutic protein libraries. The result would be rapid screening and valuable charge and isoelectric point information to aid with the development of therapeutic proteins.