STRUCTURAL REQUIREMENTS OF ANTIBODY COMBINING SITES

描述（改编自研究者摘要）： 该项目旨在探索多克隆抗体库的潜力， 治疗和/或诊断工具 癌症、微生物感染、自身免疫和免疫缺陷，以及 以预防移植排斥和移植物抗宿主病。 多克隆抗体库将结合联合收割机的优势， 多抗原决定簇（高亲合力，抗原 转义变量和效应器功能的有效调解）与 使用单克隆抗体的优点（无限量供应标准化 试剂，以及所需遗传物质的可用性 操纵）。 具有所需特异性的多克隆抗体文库的选择是 通过用于产生Fab噬菌体展示文库的技术而成为可能。 为了使这项技术适用于创建特定的完整的 IgG抗体，P.I.s实验室最近构建了双向 噬菌体展示和哺乳动物表达载体。 在这些载体中， (H)和轻（L）链转录单位处于不同的方向， 便于物理连接的变量对（V）的批量传输， 从噬菌体展示载体向哺乳动物细胞中表达V-VH区基因（VL-VH）。 表达载体，而不损失所选的VL-VH组合。 在目前的拨款申请中，研究人员建议继续 开发和优化多克隆抗体的生产技术 文库，使用人T淋巴细胞的CD 4+亚群作为模型系统。 本申请的具体目的是： Fab噬菌体展示文库（针对人CD 4 + T细胞） 来自两个不同物种（小鼠和鸡）;以产生Fab噬菌体 针对与T细胞耗尽的人的反应性选择的展示亚文库 外周血单核细胞和另外选择的亚文库 针对与CD 8 + T细胞的反应性;表达所选的多克隆 文库作为具有小鼠或鸡V的完整IgG抗体的文库 区域和小鼠或体外功能;并测试V区域 IgG文库的免疫原性。

DESCRIPTION (Adapted from Investigator's abstract): The long term goal of this project is to explore the potential of polyclonal antibody libraries as therapeutic and/or diagnostic tools in the treatment of diseases such as cancer, microbial infections, autoimmunity, and immunodeficiency, as well as in efforts to prevent transplant rejection and graft versus host disease. Polyclonal antibody libraries would combine the advantages of targeting multiple antigenic determinants (high avidity, low likelihood of antigen escape variants, and efficient mediation of effector functions) with the advantages of using monoclonal antibodies (unlimited supply of standardized reagents, and the availability of the genetic material for desired manipulations). The selection of polyclonal antibody libraries with desired specificities is made possible by the technology for generating Fab phage display libraries. To adapt this technology for the creation of specific libraries of intact IgG antibodies, the P.I.s laboratory has recently constructed bidirectional phage display and mammalian expression vectors. In these vectors, the heavy (H) and light (L) chain transcription units are in divergent orientations to facilitate the bulk transfer of physically linked pairs of variable (V) region genes (VL-VH) from the phage display vector to the mammalian expression vector, without loss of the selected VL-VH combinations. In the current grant application, The investigator proposes to continue to develop and optimize the technology for production of polyclonal antibody libraries, using as a model system the CD4+ subset of human T lymphocytes. The specific aims of the current application are: to generate polyclonal Fab phage display libraries (directed against human CD4+ T-cells) derived from two different species (mouse and chicken); to generate a Fab phage display sublibrary selected against reactivity with T-cell-depleted human peripheral blood mononuclear cells and a sublibrary additionally selected against reactivity with CD8+ T-cells; to express the selected polyclonal libraries as libraries of intact IgG antibodies with mouse or chicken V regions and mouse or functions in vitro; and to test the V region immunogenicity of the IgG libraries.