AMYLOD BETA-PEPTIDE SPECIFIC scFv ANTIBODIES

淀粉样 β 肽特异性 SCFV 抗体

• 批准号: 6754739
• 负责人: GOAR GEVORKIAN
• 金额: $ 9.72万
• 依托单位: UNIVERSITY OF MEXICO, UNAM
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-01 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6754739
• 关键词: Alzheimer' s disease; amyloid proteins; antibody; bacterial virus; chemical aggregate; combinatorial chemistry; immunologic substance development /preparation; immunotherapy; molecular cloning; neurotoxicology; passive immunization; peptide library; phage display; protein engineering; protein sequence; protein structure

DESCRIPTION (provided by applicant): Amyloid b-peptide (Ab), the major molecular component of the cerebral amyloid plaques, appears to play a central role in the neuropathology of Alzheimer's disease (AD). Compounds that prevent the formation of Ab aggregates or that selectively destroy these aggregates are attractive candidates for the development of therapeutic reagents for prevention and treatment of Alzheimer's disease. Also, compounds that interfere with the interactions of amyloid precursor protein (APP) with other factors that are involved in directing it into pathological pathway as well as those that are capable to prevent or destroy intraneuronal accumulations of Ab, are of great interest for developing therapeutic molecules for Alzheimer's disease. In this project we are proposing the possible immunological intervention for prevention and treatment of AD applying phage display technology for the construction of the first anti-Ab single chain fragment variable (scFv) antibody library and the selection of individual phage clones expressing Ab-specific scFv antibody, capable of preventing the aggregation of Ab or dissolving the preexisting aggregates, scFv phage display library enriched in anti-Ab antibodies will be constructed using the first strand cDNA synthesized from mRNA of spleen and lymph node cells isolated from mice immunized with Ab. The constructed library as well as a human scFv library will be used in bioselection procedure to isolate Ab-specific scFvs expressed on phage. The amino acid sequences of antibody complementarity-determinig regions (CDR) will be prepared and used as "mini-antibodies". Evaluation of in vitro Ab aggregation and neurotoxicity in the presence of the selected compounds: scFv antibodies expressed on phage or in soluble form and "mini-antibodies" will be performed. The most promising molecules selected in this project will be proposed for further evaluation in animal models to all researchers interested in these studies. If successful, these molecules could be of interest for passive immunization in humans, may be after modification with substances that would increase their blood-brain barrier permeability.

描述(申请人提供)：淀粉样b-肽(Ab)，大脑淀粉样斑块的主要分子成分，似乎在阿尔茨海默病(AD)的神经病理中发挥着核心作用。阻止抗体聚集体的形成或选择性地破坏这些聚集体的化合物是开发用于预防和治疗阿尔茨海默病的治疗试剂的有吸引力的候选者。此外，干扰淀粉样前体蛋白(APP)与其他因素的相互作用的化合物，以及那些能够防止或破坏抗体在神经元内积累的化合物，对于开发治疗阿尔茨海默病的分子非常有兴趣。在本项目中，我们提出了预防和治疗AD的可能的免疫干预措施，应用噬菌体展示技术构建了第一个抗单抗单链抗体(ScFv)抗体库，并筛选出能够防止抗体聚集或溶解先前存在的聚集体的表达单链抗体的噬菌体克隆，利用从经抗体免疫的小鼠的脾和淋巴结细胞中合成的第一链cDNA构建了富含抗抗体的ScFv噬菌体展示文库。所构建的单链抗体文库和人源单链抗体文库将用于生物选择过程，以分离表达于噬菌体上的抗体特异性单链抗体。抗体互补决定区(CDR)的氨基酸序列将被制备成“小抗体”。在所选化合物存在的情况下评估体外抗体聚集和神经毒性：将进行单链抗体在噬菌体上或以可溶形式表达的抗体和“迷你抗体”的研究。在这个项目中选择的最有希望的分子将被推荐给所有对这些研究感兴趣的研究人员在动物模型中进行进一步的评估。如果成功，这些分子可能会对人类的被动免疫产生兴趣，可能是在用会增加其血脑屏障通透性的物质进行修饰后。