Diverse Neutralizing Human Monoclonal Antibodies to CMV

多种抗 CMV 中和人单克隆抗体

• 批准号: 6644673
• 负责人: ANGRAY S KANG
• 金额: $ 10万
• 依托单位: AVANIR PHARMACEUTICALS
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-05-01 至 2004-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6644673
• 关键词: SCID mouse; cell line; cytomegalovirus; enzyme linked immunosorbent assay; human tissue; immunologic substance development /preparation; molecular cloning; monoclonal antibody; neutralizing antibody; polymerase chain reaction; recombinant proteins; transfection

DESCRIPTION (provided by applicant): The long-term goal of this proposal (Phase I and II) is to develop a human monoclonal antibody-based preparation for use in treating CMV infections or transmission. Commercial applications for human monoclonal antibodies are extensive. Phase I specific aims are: 1) Screen matching plasma of random buffy coats to identify CMV immune donors, and engraft selected donor cells in SCID mice (hu-PBL-SCID system). 2) Immunize with recombinant gB or gH proteins (neutralizing targets for CMV), and detect specific serum antibodies by ELISA. Optional EBV super-infection may be used. 3) Harvest cells from selected mice, and isolate antigen specific human B/plasma cells by a novel method (antibody rescue technology). Clone heavy and light chain V regions from cells by RT-PCR. 4) Isolate transiently transfected cells producing antigen-specific human monoclonal antibodies. Characterize these for epitope group, affinity, neutralizing capacity, and strain specificity. The main goal of the phase II proposal will be the configuration of at least one combination of human monoclonals for treatment of CMV infection. The phase II SBIR proposal may initially require the leneration of a larger panel of antibodies, based on the success of phase I.

描述（由申请人提供）：该提案（第一阶段和第二阶段）的长期目标是开发一种基于人单克隆抗体的制剂，用于治疗巨细胞病毒感染或传播。人单克隆抗体的商业应用非常广泛。 第一阶段的具体目标是： 1) 筛选随机血沉棕黄层匹配血浆以识别 CMV 免疫供体，并将选定的供体细胞移植到 SCID 小鼠中（hu-PBL-SCID 系统）。 2）用重组gB或gH蛋白（CMV的中和靶点）进行免疫，并通过ELISA检测特异性血清抗体。可以选择使用 EBV 重复感染。 3）从选定的小鼠中收获细胞，并通过新方法（抗体拯救技术）分离抗原特异性的人B/浆细胞。通过 RT-PCR 从细胞中克隆重链和轻链 V 区。 4) 分离产生抗原特异性人单克隆抗体的瞬时转染细胞。表征这些表位组、亲和力、中和能力和菌株特异性。 II 期提案的主要目标是配置至少一种用于治疗 CMV 感染的人单克隆抗体组合。基于第一阶段的成功，第二阶段 SBIR 提案最初可能需要产生更大的抗体组。