NINDS/UC Davis NeuroMab Hybridoma Facility

这项建议的具体目标是开发一个全面的单抗(MAbs)文库，该文库针对 在NINDS/加州大学戴维斯分校的神经单抗设施中用于大脑(即神经单抗)。这项提议是由以下需要推动的 极大地扩大这种大脑优化单抗的可获得性，用于基础、翻译和临床神经科学 研究。高通量的后基因组方法正在以快速的速度产生数据，这些方法涉及分子 脑发育、神经元可塑性以及神经和精神障碍的机制。的验证 作为进一步基础研究或治疗学发展的潜在靶点的候选基因依赖于 这些基因的蛋白质产物的特征。针对特定基因产物的单抗可以作为关键的 在大脑中表达的基因清单和对其产品如何决定大脑的洞察之间架起了桥梁 功能。然而，许多必要的试剂要么无法获得，要么在有的情况下缺乏 在哺乳动物脑中使用时的有效性和特异性。高质量、可靠的单抗的可用性 优化用于人类、非人类灵长类动物和啮齿类动物的大脑(即神经单抗)对虚拟 神经科学的所有领域。建立一个全面的NeuroMabs文库将首先进行 利用人类、小鼠和大鼠基因组计划产生的丰富数据来产生重组 和/或与神经元蛋白片段相对应的合成免疫原。这些将被用在激烈的 从相对较短的免疫中产生大量分泌免疫球蛋白的杂交瘤的免疫方案 句号。这些大型杂交瘤库将筛选那些识别同源抗原的单抗 异源细胞，然后对整个阳性池进行全面的生化和免疫组织化学检查 分析其在脑内的疗效和特异性。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免疫球蛋白制剂，对研究界来说成本非常低。这个 分泌神经单抗的杂交瘤也将免费提供。调查人员将使用这些神经单抗 测定发育中的、成人的、老年的和患病的大脑中同源抗原的存在和相对丰度， 它们的细胞和亚细胞定位、功能相关的翻译后修饰和蛋白质-蛋白质 互动。此外，NeuroMabs可能会在蛋白质的直接功能分析中找到更多的应用 诊断程序，以及作为治疗学。

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.