UC Davis/NIH NeuroMab Facility

描述(由申请人提供)：这项建议的具体目标是继续加州大学戴维斯分校/国立卫生研究院神经单抗设施的使命：开发一个针对大脑使用而优化的单抗(单抗)的综合库(即神经单抗)。这一更新仍然是由创建加州大学戴维斯分校/国立卫生研究院神经单抗设施的原始提案中阐明的需求驱动的，而且仍然需要极大地扩大这种大脑优化的单抗的可用性，用于基础、翻译和临床神经科学研究。仍然需要针对特定基因产品的高质量抗体，这些抗体是大脑中表达的基因清单与了解其产品如何决定正常和病理条件下大脑功能之间的关键桥梁。然而，许多必要的试剂要么仍然不可用，要么在有的时候缺乏有效性和特异性，特别是在用于哺乳动物的脑制剂时。对于神经科学的几乎所有领域来说，获得高质量、可靠的、针对哺乳动物大脑使用而优化的单抗(即神经单抗)是至关重要的。我们将继续致力于通过使用与神经元蛋白片段相对应的重组和/或合成免疫原，在较短的免疫期内产生大量分泌免疫球蛋白的杂交瘤，从而建立一个全面的神经单抗文库。这些大型杂交瘤细胞池将筛选出识别异种细胞中同源抗原的单抗，然后对整个阳性细胞池进行全面的生化和免疫组织化学分析，以确定其在脑内的有效性和特异性。由此产生的脑部优化的神经单抗将继续以非常低的成本提供给研究界，作为组织培养上清或纯化的免疫球蛋白制剂。研究人员将继续使用这些神经单抗来确定发育中、成人、老年和疾病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，以及蛋白质-蛋白质相互作用。此外，NeuroMabs将继续在蛋白质的直接功能分析、诊断程序和治疗学中找到更多的应用。

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.