FAB LIBRARIES TO DISCOVER VACCINE LEADS AGAINST HIV-1

描述：（改编自申请人摘要）本研究的目标 一个提议是开发一种新的方法来获得肽， 疫苗可以预防HIV-1感染。我们的策略从一个剧目开始 这些抗体是专门设计来模拟抗体的结构， 是通过用肽缀合物免疫引起的。选择的人类生殖系V 基因将编码这种“抗肽样”抗体，因此它将 模仿一个子集的“自然”剧目，通常是引起对 肽免疫原。曲目将被表达和显示为 Fab融合到丝状噬菌体，并将针对重组 HIV-1包膜蛋白寻找结合Fab。这些Fab，反过来，应该是 倾向于结合肽，并且，在免疫时，它们应该被 肽缀合物疫苗。因此，当使用此类Fab来筛选肽时 文库，“免疫原性模拟”肽应该被发现， 与Fab交叉反应，并在用作靶抗原时引发靶抗原结合Ab。 免疫原我们建议将这些概念应用于我们正在进行的研究， 肽导致疫苗，将产生广泛中和抗体， HIV-1，以下具体目标：1.噬菌体展示Fab抗体库的构建 其设计基于人类生殖系V基因， “抗肽”抗体。2.将噬菌体展示的Fab文库用于（i） 其产生针对一组“靶”肽的结合Fab的能力， （ii）那些抗原结合Fab结合蛋白抗原的能力; （iii）Fab结合肽引发Ab的能力 与相应的靶抗原结合的反应。3.电影The Fab 重组HIV- 1包膜蛋白和完整HIV-1颗粒的文库， 获得特异性HIV-1结合Fab;并表征Fab的能力 结合并中和一组HIV-1毒株和主要分离株。4. 筛选一组16个具有HIV- 1结合活性的噬菌体肽库 Fab以获得肽配体。5.确定肽配体是否引发 免疫反应识别Env蛋白并中和HIV-1。受此 我们希望开发一种新的靶向肽疫苗， HIV-1的中和位点

DESCRIPTION: (Adapted from Applicant's Abstract) The goal of this research proposal is to develop a novel means for obtaining peptides that would serve as vaccine leads against infection by HIV-1. Our strategy begins with a repertoire of antibodies that is specifically designed to mimic the structures of Abs that are elicited by immunization with peptide conjugates. Selected human germline V genes will encode this repertoire of "anti-peptide-like" Abs, so that it will mimic a subset of the "natural" repertoire that is typically elicited against peptide-immunogens. The repertoire will be expressed and displayed as Fab-fusions to filamentous phage, and will be screened against recombinant HIV-1 envelope protein to find binding Fabs. These Fabs, in turn, should be predisposed to bind peptides, and, on immunization, they should be elicited by peptide-conjugate vaccines. Thus, when such Fabs are used to screen a peptide library, "immunogenic-mimic" peptides should be found that will both cross-react with the Fabs and elicit target-antigen-binding Abs when used as immunogens. We propose to apply these concepts in our ongoing search for peptide leads for a vaccine that will produce broadly neutralizing Abs against HIV-1, following the specific aims: 1. Construct a phage-displayed Fab library whose design is based on human germline V-genes that appear in the "anti-peptide" antibodies. 2. Validate the phage-displayed Fab library for (i) its ability to produce binding Fabs against a panel of "target" peptide and protein antigens; (ii) the ability of those antigen-binding Fabs to bind to peptides, and (iii) the ability of the Fab-binding peptides to elicit Ab responses that bind to the corresponding target antigen. 3. Screen the Fab library with recombinant HIV- 1 envelope protein and intact HIV-1 particles to obtain specific HIV-1-binding Fabs; and characterize the Fabs for their ability to bind to and neutralize a panel of HIV-1 strains and primary isolates. 4. Screen a panel of 16 phage-displayed peptide libraries with the HIV- 1 binding Fabs to obtain peptide ligands. 5. Determine whether the peptide ligands elicit immune responses that recognize Env proteins and neutralize HIV-1. By this means, we hope to develop a new means of targeting peptide vaccines against neutralizing sites on HIV- l.