EPITOPE-SPECIFIC ANTIBODIES FOR ALZHEIMER'S DISEASE

阿尔茨海默病表位特异性抗体

• 批准号: 6742330
• 负责人: VICTOR A RASO
• 金额: $ 10万
• 依托单位: BOSTON BIOTECHNOLOGY CORPORATION
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-30 至 2004-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6742330
• 关键词: Alzheimer's disease; Macaca fascicularis; aging; amyloid proteins; antibody; antibody titering; antigen antibody reaction; biotechnology; epitope mapping; genetic library; genetically modified animals; immunologic assay /test; laboratory mouse; neuritic plaques; passive immunization; recombinant proteins; vaccine development; vaccines

DESCRIPTION (provided by applicant): Recent attempts at treating Alzheimer's patients with a beta-amyloid vaccine have resulted in an adverse brain inflammation response for a significant number in the trial group. The full-length 42 amino-acid beta-amyloid peptide was used as a vaccine in that study. Given the size of that molecule, it is expected to present many potential epitopes against which antibodies can be generated. It is hypothesized that antibody interaction with one or a few of those epitopes would be therapeutically beneficial while binding to other sites could have resulted in the unfortunate side effects. The proposed research will define which sites on the molecule are best to target therapeutically and which are best to avoid so that any untoward side effects are circumvented. The immunotherapy of Alzheimer's disease will become sharply focused and more consistent by using epitope specific anti-beta-amyloid scFv human antibodies for passive immunization. A large panel of beta-amyloid antigens that represent different sites on the beta-amyloid molecule has been synthesized. These will be used to screen a yeast recombinatorial human single chain Fv antibody display library of over one billion different antibodies. Unique clones, which tightly bind to specific epitopes on the beta-amyloid molecule, will be identified. Yeast expressing those select anti-beta-amyloid antibodies will be induced and the secreted single chain Fv purified via its his tag. Several precise binding assays will be used to characterize the specificity and affinity of each distinct antibody. Functional assays will be used to test the ability of the different single chain Fv antibodies to dissolve beta-amyloid aggregates in vitro. Therapeutic potential will be evaluated using a transgenic mouse model. Effective treatment will be assessed by an ability to prevent cerebral plaque formation and/or dissipate preestablished plaques in the brain. Lastly this yeast display system will provide a unique opportunity to refine those therapeutic agents through genetic engineering so they become precisely tuned to the task at hand. For example the single chain antibodies can be modified to achieve higher binding affinities, greater functionality, bivalent binding, a prolonged serum half-life or passage into the CNS.

描述（由申请人提供）：最近尝试治疗阿尔茨海默氏症患者接受β-淀粉样蛋白疫苗的尝试导致了试验组的大量数量的脑部炎症反应不良。在该研究中，全长42个氨基酸β-淀粉样蛋白肽用作疫苗。鉴于该分子的大小，预计会呈现许多可以生成抗体的潜在表位。假设与其中一个或几个表位的抗体相互作用在治疗上是有益的，而与其他部位结合可能会导致不幸的副作用。拟议的研究将定义分子上哪些位点最好以治疗靶向，哪些是最好避免的，以使任何不耐心的副作用都被规避。通过使用表位特异性抗抗淀粉样蛋白SCFV人类抗体进行被动免疫，对阿尔茨海默氏病的免疫疗法将变得更加集中，并且更加一致。 已经合成了代表β-淀粉样蛋白分子上不同位点的大型β-淀粉样抗原。这些将用于筛选酵母重组的人类单链FV抗体显示库，该抗体具有超过十亿种不同的抗体。将确定与β淀粉样蛋白分子上特定表位紧密结合的独特克隆。表达那些精选的抗β-淀粉样抗体的酵母将被诱导，并通过其标签纯化分泌的单链FV。几种精确的结合测定将用于表征每种不同抗体的特异性和亲和力。功能测定将用于测试不同单链FV抗体在体外溶解β-淀粉样蛋白聚集体的能力。将使用转基因小鼠模型评估治疗潜力。有效治疗将通过防止脑斑块形成和/或消散大脑中预先建立的斑块的能力来评估。 最后，该酵母展示系统将提供一个独特的机会，可以通过基因工程来完善这些治疗剂，从而精确地调整了手头的任务。例如，可以修改单链抗体以实现更高的结合亲和力，更大的功能，二价结合，长时间的血清半衰期或进入CNS。