NINDS/UC Davis NeuroMab Hybridoma Facility

NINDS/加州大学戴维斯分校 NeuroMab 杂交瘤设施

• 批准号: 7095057
• 负责人: James S Trimmer
• 金额: $ 66.89万
• 依托单位: UNIVERSITY OF CALIFORNIA AT DAVIS
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-15 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7095057
• 关键词: biomedical facility; biotechnology; brain; hybridomas; immunoglobulin G; immunologic substance development /preparation; immunologic substance resource /registry /referral center; immunotherapy; molecular cloning; monoclonal antibody

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.

描述（由申请人提供）： 该提案的具体目的是开发综合的单克隆抗体（mAB）库（mAB），可在Ninds/UC Davis Neuromab设施中优化用于大脑（即神经瘤）的使用。该建议是由大大扩展此类脑部优化mAB的可用性以用于基本，转化和临床神经科学研究的驱动的。数据是从高通量后的基因组方法快速生成的，这些方法解决了脑发育，神经元可塑性以及神经系统和精神疾病的分子机制。验证候选基因作为进一步基础研究的潜在靶标或疗法的发展依赖于这些基因蛋白质产物的表征。针对已定义的消失产品的mAb可以用作大脑中表达的基因库存之间的关键桥梁，并洞悉其产品如何确定大脑功能。但是，许多必要的试剂要么不可用，要么在可用时缺乏疗效和特异性时，在哺乳动物大脑中使用。用于在人，非人类灵长类动物和啮齿动物大脑（即神经瘤）中使用的高质量，可靠的单克隆抗体的可用性至关重要。首先利用从人，小鼠和大鼠基因组项目中产生的大量数据来产生重组和/或合成免疫原子，将追求全面的神经瘤图书馆，从而追求全面的神经单抗库。这些将用于强烈的免疫协议中，该方案从相对较短的免疫期中产生大量分泌IgG的杂交瘤。这些大型杂交瘤池将被筛选为那些识别异源细胞中同源抗原的mAB，然后将整个阳性池经过全面的生物化学和免疫组织化学分析，这些分析对其在大脑中的疗效和特异性进行了。最终的大脑优化神经瘤将以非常低的成本作为组织培养物上清液或浓缩IgG制剂提供。神经瘤分泌的杂交瘤也将免费提供。研究者将使用这些神经毛电图来确定同源抗原在发育，成人，老年和患病的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以及蛋白质 - 蛋白质蛋白相互作用。此外，神经单抗可能在蛋白质，诊断程序和作为治疗剂的直接功能分析中发现其他I应用。