NINDS/UC Davis NeuroMab Hybridoma Facility

该提案的具体目的是开发针对以下优化的单克隆抗体（mAb）的综合文库： 在NINDS/UC Davis NeuroMab Facility的脑中使用（即NeuroMab）。这一提议是出于以下需要： 极大地扩展了这种脑优化mAb在基础、转化和临床神经科学中的可用性 research.高通量后基因组方法正在快速生成数据， 脑发育机制、神经元可塑性以及神经和精神疾病。的验证 候选基因作为进一步基础研究或开发治疗方法的潜在靶点，依赖于 这些基因的蛋白质产物的表征。针对特定基因产物的单克隆抗体可以作为关键的 在大脑中表达的基因清单与其产物如何决定大脑之间的桥梁 功能然而，许多必要的试剂要么无法获得，要么在可获得的情况下缺乏足够的试剂。 当用于哺乳动物脑时的功效和特异性。高质量、可靠的mAb的可用性， 优化用于人类、非人类灵长类动物和啮齿类动物脑（即NeuroMabs）是最重要的， 神经科学的所有领域。NeuroMab的综合文库的产生将通过首先采取 利用来自人类、小鼠和大鼠基因组计划的大量数据， 和/或对应于神经元蛋白片段的合成免疫原。这些将用于一个激烈的 从相对短的免疫产生大量IgG分泌杂交瘤的免疫方案 期将筛选这些大杂交瘤池中识别同源抗原的那些mAb， 异源细胞，然后对整个阳性池进行全面的生化和免疫组化 分析其在脑中的功效和特异性。由此产生的大脑优化的NeuroMabs将在 作为组织培养上清液或作为浓缩的IgG制剂，对于研究界来说成本非常低。的 分泌NeuroMab的杂交瘤也将免费提供。研究人员将使用这些NeuroMab进行 确定同源抗原在发育中的、成年的、老年的和患病的脑中的存在和相对丰度， 它们的细胞和亚细胞定位，功能相关的翻译后修饰，以及蛋白质-蛋白质 交互.此外，NeuroMabs可能在蛋白质的直接功能分析中找到其他应用， 诊断程序和治疗方法。

The specific aim of this proposal is to develop a comprehensive library of monoclonal antibodies (mAbs) optimized for use in the brain (i.e. NeuroMabs) at the NINDS/UC Davis NeuroMab Facility. This proposal is driven by the need to greatly expand the availability of such brain-optimized mAbs for use in basic, translational and clinical neuroscience research. Data are being generated at a rapid rate from high throughput post-genomic approaches addressing molecular mechanisms of brain development, neuronal plasticity, and neurological and psychiatric disorders. The validation of candidate genes as potential targets for further basic research, or for the development of therapeutics, relies on characterization of the protein products of these genes. MAbs against defined gene products can serve as the crucial bridge between the inventory of genes expressed in the brain, and insights into how their products determine brain function. However, many of the necessary reagents are either unavailable, or when available suffer from a lack of efficacy and specificity when used in mammalian brain. The availability of high-quality, reliable mAbs that have been optimized for use in human, non-human primate, and rodent brain (i.e. NeuroMabs) is of utmost importance to virtually all areas of neuroscience. The generation of a comprehensive library of NeuroMabs will be pursued by first taking advantage of the wealth of data emerging from the human, mouse and rat genome projects to generate recombinant and/or synthetic immunogens corresponding to fragments of neuronal proteins. These will be used in an intense immunization protocol that yields large numbers of IgG-secreting hybridomas from a relatively short immunization period. These large hybridoma pools will be screened for those mAbs that recognize the cognate antigen in heterologous cells, and then the entire positive pool subjected to comprehensive biochemical and immunohistochemical analyses of their efficacy and specificity in brain. The resultant brain-optimized NeuroMabs will be made available at very low cost to the research community as tissue culture supernatants or as concentrated IgG preparations. The NeuroMab secreting hybridomas will also be made freely available. Investigators will use these NeuroMabs for determining the presence and relative abundance of the cognate antigens in developing, adult, aged, and diseased brain, their cellular and subcellular localization, functionally relevant post-translational modifications, and protein-protein interactions. Moreover, NeuroMabs may find additional applications in direct functional analyses of proteins, in diagnostic procedures, and as therapeutics.