STRUCTURAL ENCAPSULATION OF MODEL PROTEINS IN BIOPOLYMER

描述（改编自应用）：许多蛋白质，包括 药物相关的，可以提供延长的时间， 合成生物相容性聚合物对人体的影响。这些发现认为， 有望显著扩大此类蛋白质用于预防 和治愈疾病。然而，在生物相容性中蛋白质的包封是不可能的。 聚合物包括对其结构有害的事件，例如暴露于 有机溶剂、剪切力、疏水界面和脱水。而 分析蛋白质从这种装置的释放动力学是常规的， 申请人最近提供了关于蛋白质的第一个结构数据， 生物相容性聚合物。本提案的总体目标是， 新开发的光谱方法，以进一步合理化的影响， 包埋条件对模型蛋白结构的影响及相关研究 结构与释放和稳定性数据。最终目的是了解 蛋白质包封的结构和稳定性结果 生物相容性聚合物，以提供合理的配制策略。为此 他们将主要采用傅里叶变换红外（FTIR）光谱， 定量表征模型二级结构的变化 蛋白质在其封装的不同阶段。此外，高分辨率 H/D交换NMR光谱将用于获得互补结构 数据结构数据将与稳定性参数相关联，例如 从装置中累积释放的蛋白质及其特异性 生物活性他们将特别专注于优化 非水包封方法，其中他们已经实现了 两种模型蛋白（重组人生长激素）结构保存 和牛血清白蛋白）， 聚乳酸-羟基乙酸共聚物。此外，最常用的方法是 蛋白质封装，即双乳化溶剂蒸发技术，将 通过工艺参数的变化进行系统分析。他们是 特别感兴趣的是稳定蛋白质在有害的 第一封装步骤，其中形成水-有机溶剂界面 并且蛋白质溶液经受高剪切力。他们还将 解决构象问题。蛋白质在有机溶剂中的稳定性 实验上，特别是专注于添加剂对它们的影响， 热稳定性

Description (Adapted from Application): Many proteins, including pharmaceutically relevant ones, can be delivered for prolonged times from synthetic biocompatible polymers to the human body. These findings hold the promise of significantly expanding the use of such proteins for the prevention and cure of diseases. However, the encapsulation of proteins in biocompatible polymers includes events detrimental to their structure, such as exposure to organic solvents, shear forces, hydrophobic interfaces, and dehydration. While it is routine to analyze the release kinetics of proteins from such devices, the applicants recently provided the first structural data on proteins within biocompatible polymers. The overall objective of this proposal is to apply the newly developed spectroscopic methods to further rationalize the impact of the encapsulation conditions on the structure of model proteins and relate structural with release and stability data. The ultimate goal is to understand the structure and stability consequences of protein encapsulation in biocompatible polymers to provide rational formulation strategies. To this end they will primarily employ Fourier-transform infrared (FTIR) spectroscopy to quantitatively characterize changes in the secondary structure of model proteins at various stages of their encapsulation. In addition, high-resolution H/D exchange NMR spectroscopy will be applied to gain complementary structural data. The structural data will be correlated with stability parameters, such as cumulative release of the proteins from the devices and their specific biological activity. They will, in particular, concentrate on optimizing non-aqueous encapsulation approaches, where they have already achieved structural preservation of two model proteins (recombinant human growth hormone and bovine serum albumin) upon encapsulation in FDA-approved poly(lactic-co-glycolic)acid. In addition, the most common method used for protein encapsulation, the double emulsion solvent evaporation technique, will be systematically analyzed by variation of the processing parameters. They are particularly interested in the stabilization of proteins in the detrimental first encapsulation step, where an aqueous-organic solvent interface is formed and protein solutions are subjected to high shear forces. They will also address the conformational. Stability of proteins in organic solvents experimentally, particularly focussing on the influence of additives on their thermal stability.