NINDS/UC Davis NeuroMab Hybridoma Facility

描述（由申请人提供）： 该提案的具体目的是在NINDS/UC Davis NeuroMab Facility开发一个针对脑（即NeuroMab）使用优化的单克隆抗体（mAb）的综合文库。这一提议是由于需要大大扩展这种脑优化mAb的可用性，以用于基础，转化和临床神经科学研究。高通量后基因组方法正在快速生成数据，这些方法涉及大脑发育、神经元可塑性以及神经和精神疾病的分子机制。候选基因作为进一步基础研究或开发治疗药物的潜在靶点的验证依赖于这些基因的蛋白质产物的表征。针对已定义的Gone产物的单克隆抗体可以作为大脑中表达的基因清单与了解其产物如何决定大脑功能之间的关键桥梁。然而，许多必要的试剂要么是不可用的，要么是在哺乳动物脑中使用时缺乏有效性和特异性。已优化用于人类、非人类灵长类动物和啮齿类动物脑的高质量、可靠的mAb（即NeuroMab）的可用性对神经科学的所有领域都至关重要。将通过首先利用从人、小鼠和大鼠基因组计划中出现的大量数据来产生对应于神经元蛋白片段的重组和/或合成免疫原，来追求NeuroMab的综合文库的产生。这些将用于从相对短的免疫期产生大量IgG分泌杂交瘤的强化免疫方案。将筛选这些大杂交瘤池中识别异源细胞中同源抗原的mAb，然后对整个阳性池进行全面的生物化学和免疫组织化学分析，以确定其在脑中的有效性和特异性。所得到的脑优化的NeuroMabs将以非常低的成本作为组织培养上清液或浓缩的IgG制剂提供给研究界。分泌NeuroMab的杂交瘤也将免费提供。研究者将使用这些NeuroMabs确定发育中、成人、老年和患病脑中同源抗原的存在和相对丰度、其细胞和亚细胞定位、功能相关的翻译后修饰和蛋白质-蛋白质相互作用。此外，NeuroMabs可能在蛋白质的直接功能分析、诊断程序和治疗中找到额外的应用。

DESCRIPTION (provided by applicant): 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 gone 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 virtual 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 i applications in direct functional analyses of proteins, in diagnostic procedures, and as therapeutics.