Biotechnology Research Partnerships Renewal: Aggregation of Therapeutic Proteins

描述(申请人提供)：治疗性蛋白质为多种人类疾病和障碍提供独特的治疗方法，包括癌症、多发性硬化症、血友病、类风湿性关节炎、克罗恩病和糖尿病。不幸的是，在接受治疗性蛋白产品(高达50%或更高)的患者中，很大一部分患者由于对该产品的免疫反应而失去疗效。蛋白质聚集体的存在可以刺激免疫原性。在这个项目中，我们将使用一种分层的方法，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的分子水平信息。实验将利用具有设计化学特性的合成蛋白质、小鼠模型中的治疗蛋白模型和商业治疗蛋白。将使用一系列广泛的实验技术来分析蛋白质聚集体及其形成途径，包括单分子全内反射荧光-Forster共振能量转移、小角中子散射、小角x射线散射微流成像、表面微流变学、层析、圆二色光谱和红外光谱。这项更新提案的一个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蛋白质聚集，蛋白质聚集是由于与输送设备(例如注射器、静脉输液袋、静脉输液管路和输送泵)中的接口相互作用而发生的。

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).