Conformational antibodies recognizing amyloid epitopes

描述（由申请人提供）：许多神经系统疾病和其他疾病与淀粉样原纤维蛋白聚集体的形成有关。虽然淀粉样蛋白的蛋白质成分因不同的疾病而不同，但淀粉样蛋白的三维形状似乎在所有情况下都是相同的。事实证明，获得淀粉样蛋白的详细结构信息极其困难，这影响了我们理解和设计治疗阿尔茨海默病（AD）、帕金森氏病、亨廷顿氏病以及一系列其他毁灭性疾病的努力。我们已经开发出了抗体分子，它具有与淀粉样蛋白原纤维结合的独特能力，而不管组成蛋白质的氨基酸序列如何。在这里，我们建议开发这些构象抗体作为工具，以提高我们对淀粉样蛋白结构及其在人类疾病中如何发展的理解。具体目标是：(1)开发额外的抗体和抗体片段，(2)在结构上表征这些抗体作为淀粉样蛋白原纤维结构的新窗口，(3)表征抗体-淀粉样蛋白相互作用的基本基础，(4)开发抗体用于检测组织和血清中的淀粉样蛋白，以及在体外淀粉样蛋白形成过程中监测淀粉样蛋白基序的出现。这项工作涉及的工具将包括杂交瘤和噬菌体展示技术，重组表达，诱变，蛋白质建模和晶体学，免疫化学结合测定，以及体外淀粉样纤维组装反应。由于最近描述的淀粉样蛋白疾病（如AD）的疫苗方法依赖于抗体的产生，因此表征抗淀粉样蛋白反应的结构基础非常重要。了解淀粉样蛋白的结构和这些原纤维的生长方式也很重要，这是开发损害原纤维细胞毒性和原纤维生长的药物的一种手段。最后，淀粉样蛋白作为一种普遍存在的蛋白质聚合物的替代折叠模式，其结构对我们理解生命的分子基础至关重要。

DESCRIPTION (provided by applicant): A number of neurological and other diseases are associated with the formation of protein aggregates called amyloid fibrils. Although the protein component of amyloid is different for different diseases, the three-dimensional shape of the amyloid seems to be the same in all cases. It has proven exceedingly difficult to get detailed structural information on amyloid, compromising our efforts to understand and devise treatments for Alzheimer's disease (AD), Parkinson's disease, Huntington's disease, and an array of other devastating conditions. We have developed antibody molecules that have the unique ability to bind to amyloid fibrils, regardless of the amino acid sequence of the constituent protein. Here we propose to develop these conformational antibodies as tools to improve our understanding of the amyloid structure and how it develops in human diseases. The specific aims are to (1) develop additional antibodies and antibody fragments, (2) structurally characterize these antibodies as a novel window onto the structure of amyloid fibrils, (3) characterize the fundamental basis of the antibody-amyloid interaction, and (4) develop the antibodies into tests for detecting amyloid in tissue and serum, as well as to monitor the emergence of the amyloid motif during in vitro amyloidogenesis. The tools involved in this work will include hybridoma and phage display technology, recombinant expression, mutagenesis, protein modeling and crystallography, immunochemical binding assays, and in vitro amyloid fibril assembly reactions. Since recently described vaccine approaches to amyloid diseases such as AD depend on the generation of antibodies, characterizing the structural basis of the anti-amyloid reaction is important. It is also important to understand the structure of amyloid and how these fibrils grow, as a means toward developing agents that will compromise fibril cytotoxicity and fibril growth. Finally, as a ubiquitous alternative folding pattern of protein polymers, amyloid and its structure is of fundamental importance to our understanding of the molecular basis of life.