Conformation-specific Single-chain Antibodies as Neurodegeneration Research Tools

描述(申请人提供)：这个开发项目的具体目标是优化和表征构象特异性单链抗体(ScFv)，作为识别和操纵细胞内错误折叠的神经变性蛋白的工具。我们已经开发出独特的能力，使我们能够产生针对特定蛋白质形态的单链抗体，并在细胞内测试它们的功能。鉴于阿尔茨海默氏症、帕金森氏症、亨廷顿病、普恩病毒和ALS疾病所涉及的蛋白质在结构上的共性，这些试剂将使研究这些不同疾病的研究人员能够在细胞环境中区分寡聚体和纤维状聚集体。它还将允许将特定形式的蛋白质重定向到其他细胞内隔间，以测试最关键的作用部位所在的假设。 这是一个多PI应用程序。亚利桑那州立大学Michael Sierks博士的实验室目前正在挑选抗α-突触核蛋白寡聚体和抗纤维单链抗体。第一年的大部分时间将由纽约州奥尔巴尼Wadsworth中心的Anne Messer博士致力于高效快速筛选神经元细胞系的细胞内表达，随后将立即对最有希望的单链抗体进行更劳动密集型的亲和力和稳定性工程。到第二年，大部分工作将包括在奥尔巴尼完成的详细生物学研究，以及亚利桑那州立大学需要的进一步蛋白质相互作用和优化。公共卫生：随着我们人口的老龄化，神经退行性疾病正在给人类和经济造成越来越大的损失。工程抗体疗法已经用于癌症和传染病，并为神经系统疾病药物发现和治疗提供了一个令人兴奋的新平台。该项目专门针对帕金森病中蛋白质错误折叠的后果，直接应用于相关疾病，如阿尔茨海默氏症、亨廷顿氏症和肌萎缩侧索硬化症。

DESCRIPTION (provided by applicant): The specific goal of this developmental project is to optimize and characterize conformation-specific single-chain Fv (scFv) antibodies as tools to recognize and manipulate misfolded neurodegeneration proteins intracellularly. We have developed unique capabilities that enable us to generate scFvs that target specific protein morphologies and to test their function intracellularly. Given the known structural commonality of proteins involved in Alzheimer's, Parkinson's, Huntington's, prion, and ALS diseases, these reagents will allow investigators working on these various diseases to distinguish between oligomeric and fibrillar forms of aggregates in a cellular context. It will also allow redirection of specific forms of proteins to other intracellular compartments to test hypotheses of where the most critical site of action resides. This is a multiple-PI application. The anti-alpha-synuclein oligomeric and anti- fibrillar scFvs are currently being selected in the laboratory of Dr. Michael Sierks, at ASU. Much of the first year will be devoted to efficient rapid screening of intracellular expression in neuronal cell lines by Dr. Anne Messer, Wadsworth Center in Albany, NY, followed immediately by the more labor-intensive engineering of affinity and stability of the most promising scFvs. By the second year, the majority of the work will consist of detailed biological studies done in Albany, with further protein interactions and optimization as needed at ASU. PUBLIC HEALTH REVELANCE: Neurodegenerative diseases are taking an increasing human and financial toll as our population ages. Engineered antibody therapeutics are already in use for cancer and infectious disease, and offer an exciting new platform for neurological disease drug discovery and therapeutics. This project specifically targets the consequences of protein misfolding in Parkinson's disease, with direct applications to related diseases such as Alzheimer's, Huntington's, and ALS.