UC Davis/NIH NeuroMab Facility

描述（由申请方提供）：本提案的具体目的是继续UC Davis/NIH NeuroMab Facility的使命：开发针对脑（即NeuroMab）优化的单克隆抗体（mAb）的综合文库。这种更新仍然是由需求驱动的，在最初的提议中阐明了创建加州大学戴维斯分校/NIH NeuroMab设施，并且仍然存在，以大大扩展这种脑优化mAb的可用性，用于基础，转化和临床神经科学研究。仍然需要针对确定的基因产物的高质量抗体，所述抗体充当在脑中表达的基因的清单与理解其产物如何在正常和病理条件下决定脑功能之间的关键桥梁。然而，许多必要的试剂仍然不可用，或者当可用时缺乏功效和特异性，特别是当用于哺乳动物脑制备时。获得高质量、可靠的单克隆抗体（即NeuroMab），这些抗体已被优化用于哺乳动物大脑，这对神经科学的几乎所有领域都至关重要。我们将继续追求通过使用重组和/或合成的免疫原对应于神经元蛋白的片段，在一个强烈的免疫方案，产生大量的IgG分泌杂交瘤从一个相对较短的免疫期的NeuroMabs的综合性库的生成。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。由此产生的脑优化的NeuroMabs将继续以非常低的成本作为组织培养上清液或纯化的IgG制剂提供给研究界。研究者将继续使用这些NeuroMabs来确定同源抗原在发育、成人、老年和患病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，NeuroMabs将继续在蛋白质的直接功能分析，诊断程序和治疗方面找到更多的应用。 公共卫生相关性：为哺乳动物大脑使用而优化的高质量、可靠的mAb的可用性对基础、转化和临床神经科学的几乎所有领域都至关重要。

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.