UC Davis/NIH NeuroMab Facility

描述（由申请人提供）：本提案的具体目的是继续UC Davis/NIH NeuroMab设施的使命：开发一个优化用于大脑的单克隆抗体（mab）的综合文库（即NeuroMab）。这种更新仍然是由需求驱动的，在最初创建加州大学戴维斯分校/美国国立卫生研究院神经单克隆抗体设施的提议中明确表达，并且仍然存在，以极大地扩大这种大脑优化的单克隆抗体的可用性，用于基础，转化和临床神经科学研究。目前仍然需要针对特定基因产物的高质量抗体，这些抗体作为大脑中表达的基因清单和理解其产物如何决定正常和病理条件下的大脑功能之间的关键桥梁。然而，许多必要的试剂要么没有，要么即使有，也缺乏效力和特异性，特别是在哺乳动物脑制剂中使用时。高质量，可靠的单克隆抗体的可用性已经优化用于哺乳动物的大脑（即神经单克隆抗体）是至关重要的，几乎所有的神经科学领域。我们将继续通过使用重组和/或合成的与神经元蛋白片段相对应的免疫原，在一个相对较短的免疫周期内产生大量分泌igg的杂杂瘤，以建立一个全面的神经单抗文库。在这些大型杂交瘤池中筛选识别异种细胞同源抗原的单克隆抗体，然后对整个阳性池进行生化和免疫组织化学综合分析其在脑中的有效性和特异性。由此产生的脑优化神经单克隆抗体将继续以极低的成本作为组织培养上清或纯化的IgG制剂提供给研究界。研究人员将继续使用这些neuromab来确定同源抗原在发育、成人、老年和患病大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和蛋白质-蛋白质相互作用。此外，neuromab将继续在蛋白质的直接功能分析、诊断程序和治疗方面寻找更多的应用。

DESCRIPTION (provided by applicant): The specific aim of this proposal is to continue the mission of the UC Davis/NIH NeuroMab Facility: to develop a comprehensive library of monoclonal antibodies (mAbs) optimized for use in the brain (i.e. NeuroMabs). This renewal remains driven by the need, articulated in the original proposal to create the UC Davis/NIH NeuroMab Facility, and that still remains, to greatly expand the availability of such brain-optimized mAbs for use in basic, translational and clinical neuroscience research. There remains a need for high- quality antibodies against defined gene products that serve as the crucial bridge between the inventory of genes expressed in the brain, and understanding how their products determine brain function in normal and pathological conditions. However, many necessary reagents remain either unavailable, or when available suffer from a lack of efficacy and specificity, especially when used in mammalian brain preparations. The availability of high-quality, reliable mAbs that have been optimized for use in mammalian brain (i.e. NeuroMabs) is of utmost importance to virtually all areas of neuroscience. We will continue to pursue the generation of a comprehensive library of NeuroMabs by using recombinant and/or synthetic immunogens corresponding to fragments of neuronal proteins 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 continue to be made available at very low cost to the research community as tissue culture supernatants or as purifed IgG preparations. Investigators will continue to 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 will continue to find additional applications in direct functional analyses of proteins, in diagnostic procedures, and as therapeutics. PUBLIC HEALTH RELEVANCE: The availability of high-quality, reliable mAbs that have been optimized for use in mammalian brain is of utmost importance to virtually all areas of basic, translational and clinical neuroscience.