Biotechnology Research Partnerships Renewal: Aggregation of Therapeutic Proteins

描述（由申请人提供）：治疗性蛋白质为许多人类疾病和失调提供独特的治疗方法，包括癌症、多发性硬化症、血友病、类风湿关节炎、克罗恩病和糖尿病。不幸的是，在很大一部分接受治疗性蛋白质产品的患者中（高达50%或更高），由于对产品的免疫反应，疗效丧失。免疫原性可能被蛋白质聚集物的存在所刺激。在这个项目中，我们将使用分层方法，使用先进的计算机模拟与最先进的实验相结合，提供关于蛋白质聚集发生机制的详细分子水平信息。实验将利用具有设计化学特性的合成蛋白，在小鼠模型中模拟治疗蛋白和商业治疗蛋白。蛋白质聚集体及其形成途径将使用广泛的实验技术进行分析，包括单分子全内反射荧光-福斯特共振能量转移，小角度中子散射，小角度x射线散射微流成像，表面微流变学，色谱学，圆二色光谱和红外光谱。该更新提案的主要重点将是将我们目前在均相溶液中蛋白质聚集的工作扩展到由于与界面（例如在输送设备中发现的界面）相互作用而发生的蛋白质聚集（例如，注射器，静脉输液袋，静脉输液管和输送泵）。

DESCRIPTION (provided by applicant): Therapeutic proteins provide unique treatments for numerous human diseases and disorders including cancer, multiple sclerosis, hemophilia, rheumatoid arthritis, Crohn's disease and diabetes. Unfortunately, in significant fractions of patients receiving therapeutic protein products (up to 50% or higher), efficacy is lost due to immune response to the product. Immunogenicity may be stimulated by the presence of protein aggregates. In this project, we will use a hierarchical approach that uses advanced computer simulations combined with state-of-the art experiments to provide detailed, molecular-level information on the mechanisms by which protein aggregation occurs. Experiments will utilize synthetic proteins with designed chemical characteristics, model therapeutic proteins in murine models and commercial therapeutic proteins. Protein aggregates and pathways for their formation will be analyzed using a wide array of experimental techniques including single-molecule total internal reflectance fluorescence-Forster resonant energy transfer, small angle neutron scattering, small angle x-ray scattering micro-flow imaging, surface micro-rheology, chromatography, circular dichroism spectroscopy and infrared spectroscopy. A major focus of this renewal proposal will be the extension of our current work on protein aggregation in homogeneous solutions to protein aggregation that occurs due to interactions with interfaces such as those found in delivery devices (e.g., syringes, IV bags, IV lines, and delivery pumps).