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病症提供独特的治疗，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，在接受治疗性蛋白质产品的患者的显著部分（高达50%或更高）中，由于对产品的免疫应答而丧失功效。免疫原性可以通过蛋白质聚集体的存在来刺激。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结合最先进的实验，以提供详细的，分子水平的信息，蛋白质聚集发生的机制。实验将利用具有设计化学特征的合成蛋白质、鼠模型中的模型治疗蛋白质和商业治疗蛋白质。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，小角中子散射，小角X射线散射微流成像，表面微流变学，色谱，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关于均匀溶液中蛋白质聚集的工作扩展到蛋白质聚集，这种蛋白质聚集是由于与界面的相互作用而发生的，如在递送装置中发现的那些（例如，注射器、IV袋、IV管线和输送泵）。

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).