RECOMBINANT ANTIBODIES FOR INFANT PROTECTION

用于婴儿保护的重组抗体

• 批准号: 6373507
• 负责人: Sherie L Morrison
• 金额: $ 26.58万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-30 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6373507
• 关键词: Escherichia coli; Escherichia coli infections; agglutination reaction; antibacterial antibody; antibody receptor; cell adhesion; cell mediated cytotoxicity; chickens; complement; enterotoxins; genetically modified animals; immunoglobulin A; immunoglobulin G; intravenous administration; laboratory mouse; microorganism disease chemotherapy; monoclonal antibody; mucosal immunity; neutralizing antibody; nonhuman therapy evaluation; oral administration; phagocytes; pharmacokinetics; recombinant proteins; secretory protein

DESCRIPTION: (Adapted from the Investigator's abstract): As recombinant antibodies (Abs) are increasingly used as therapeutic agents for the treatment of infectious disease and malignancy, it becomes critically important that we understand the mechanisms through which antibodies provide protection. Armed with this information, we will be able to produce the most effective Abs. It is also important that we are able to produce recombinant Abs at a cost that will make them broadly available. In the proposed experiments the investigators will address these issues. They will attempt to produce IgA with improved functional properties. IgA is the principal Ab providing protection in the hostile environment of the mucosal surface where it is susceptible to proteolytic attack and one goal will be to identify and alter the positions on sIgA that are most susceptible to proteolysis. These experiments should both yield a more stable sIgA and also give insights into its structure. The biologic properties of IgA suggest that it may be the ideal antibody for some parenteral applications. However, serum IgA is rapidly cleared. They will attempt to increase the serum half-life of IgA by providing it with an FcRn binding site and by identifying and removing the N-carbohydrate moiety responsible for the rapid clearance of IgA2 by the asialoglycoprotein receptor. These studies will provide IgA with a longer half-life and give additional insights into the mechanisms and pathways of Ab clearance. Complement activation can lead to the elimination of pathogens by both cytolysis and opsonization. They will attempt to produce IgA with the ability to activate complement. These experiments will provide an IgA with the ability to activate the inflammatory cascade and will also further our understanding of the sequences on the Ab that mediate interaction with the complement proteins. They will attempt to define the mechanisms of Ab-mediated protection using the suckling mouse model for enterotoxigenic Escherichia coli (ETEC) and monoclonal antibodies against the F41 antigen. In vitro activity in assays such as adherence, agglutination, complement-mediated cytotoxicity and stability will be compared with the ability to provide in vivo protection. As one step to our long-term goal of using the chicken as a bioreactor for Ab production, they will further define the fine specificity of the receptor responsible for Ab transport with the ultimate goal of cloning it. Definition of the sequences required for transport receptor recognition should make it possible to design Abs that are effectively transported. It also may be possible to use this sequence to target other proteins to the chicken egg. Cloning of the receptor will define its structure and allow us to fully characterize this novel transport system.

描述：（改编自研究者的摘要）：作为重组体 抗体（Abs）越来越多地用作治疗药物 面对传染病和恶性肿瘤，我们必须 了解抗体提供保护的机制。武装 有了这些信息，我们将能够生产出最有效的 Abs。这是 同样重要的是我们能够以一定的成本生产重组抗体 使它们广泛可用。在拟议的实验中，研究人员将 解决这些问题。他们将尝试生产具有改进功能的 IgA 特性。 IgA 是在敌对环境中提供保护的主要抗体 粘膜表面易被蛋白水解的环境 攻击，一个目标是识别并改变 sIgA 上的位置， 最容易被蛋白水解作用。这些实验应该都会产生更多 稳定的 sIgA 并深入了解其结构。生物学特性 IgA 的含量表明它可能是某些肠外注射的理想抗体 应用程序。然而，血清IgA很快被清除。他们将尝试 通过提供 FcRn 结合位点来延长 IgA 的血清半衰期 并通过识别和去除负责的N-碳水化合物部分 脱唾液酸糖蛋白受体快速清除 IgA2。这些研究将 为 IgA 提供更长的半衰期，并提供更多关于 抗体清除的机制和途径。补体激活可导致 通过细胞溶解和调理作用消除病原体。他们将尝试 产生具有激活补体能力的IgA。这些实验将 提供具有激活炎症级联反应能力的 IgA，并将 也进一步加深了我们对介导抗体序列的理解 与补体蛋白的相互作用。他们将尝试定义 使用乳鼠模型研究抗体介导的保护机制 产肠毒素大肠杆菌 (ETEC) 和针对该菌株的单克隆抗体 F41抗原。粘附、凝集等测定中的体外活性 补体介导的细胞毒性和稳定性将与 提供体内保护的能力。作为实现我们长期目标的第一步 使用鸡作为抗体生产的生物反应器，他们将进一步定义 负责抗体转运的受体的精细特异性 最终目标是克隆它。传输所需序列的定义 受体识别应该能够设计出有效的抗体 运输。也可以使用该序列来靶向其他 鸡蛋中的蛋白质。受体的克隆将定义其结构 并使我们能够充分描述这种新颖的运输系统。