Conformation-specific Single-chain Antibodies as Neurodegeneration Research Tools

构象特异性单链抗体作为神经变性研究工具

基本信息

批准号：
7480889
负责人：
ANNE MESSER
金额：
$ 17.64万
依托单位：
ARIZONA STATE UNIVERSITY-TEMPE CAMPUS
依托单位国家：
美国
项目类别：
财政年份：
2008
资助国家：
美国
起止时间：
2008-04-01 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7480889
关键词：
Adopted Affinity Age Alzheimer&apos s Disease Amyloid beta-Protein Amyotrophic Lateral Sclerosis Antibodies Antigen Targeting Atomic Force Microscopy Binding Biological Biological Assay Biological Models Cell Line Cell Nucleus Cell model Cells Communicable Diseases Condition Development Disease Disease model Drosophila genus Engineering Future Generations Goals Human Huntington Disease Image Immunoglobulin Fragments Immunoglobulin G In Vitro Investigation Laboratories Length Libraries Location Malignant Neoplasms Methods Modeling Molecular Conformation Morphology Nerve Degeneration Neurodegenerative Disorders Neurons Nuclear Numbers Pan Genus Paper Parents Parkinson Disease Pathogenesis Phage Display Play Population Prion Diseases Prions Protein Overexpression Proteins Protocols documentation Public Health Publications Publishing Purpose Range Reagent Recombinant Antibody Research Research Personnel Role Screening procedure Site Specificity Technology Testing Therapeutic Therapeutic antibodies Toxic effect Transgenes Variant Work alpha synuclein antibody engineering base concept cross reactivity direct application drug discovery experience human Huntingtin protein improved in vivo mouse model nervous system disorder protein misfolding research and development tool

项目摘要

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.

描述（由申请人提供）：此发展项目的具体目标是优化和表征特定于构象的单链FV（SCFV）抗体，作为识别和操纵错误折叠神经变性蛋白的工具。我们开发了独特的功能，使我们能够生成靶向特定蛋白质形态并细胞内测试功能的SCFV。鉴于涉及阿尔茨海默氏症，帕金森氏症，亨廷顿，prion和ALS疾病的蛋白质的已知结构共同点，这些试剂将允许研究这些各种疾病的研究人员在细胞上下文中区分聚集体的寡聚和纤维状形式。它还将允许将特定形式的蛋白质重定向到其他细胞内室，以测试最关键的作用部位所在位置的假设。这是一个多重PI应用程序。目前正在ASU的Michael Sierks博士的实验室中选择了抗α-核蛋白低聚和抗纤维SCFV。第一年的大部分时间将致力于有效地筛查纽约州奥尔巴尼市沃兹沃思中心的安妮·梅塞尔（Anne Messer）博士在神经元细胞系中的细胞内表达，然后立即进行了更有希望的SCFV的劳动密集型工程和稳定性。到第二年，大多数工作将包括在奥尔巴尼进行的详细生物学研究，并根据ASU的需要进行进一步的蛋白质相互作用和优化。公共卫生的启示：随着我们人口的年龄，神经退行性疾病正在越来越多的人类和经济损失。工程抗体疗法已经用于癌症和传染病，并为神经疾病的药物发现和疗法提供了一个令人兴奋的新平台。该项目专门针对帕金森氏病中蛋白质折叠率误后的后果，直接应用于相关疾病，例如阿尔茨海默氏症，亨廷顿和ALS。