Antigen-microarray validated monoclonal antibody library for analysis of brain-ex

用于脑前分析的抗原微阵列验证单克隆抗体库

• 批准号: 8000889
• 负责人: Seth Blackshaw
• 金额: $ 29.09万
• 依托单位: CDI LABORATORIES, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-05 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8000889
• 关键词: Advertising; Affinity; Antibodies; Antibody Specificity; Antigens; Binding; Biology; Brain; Cataloging; Catalogs; Cells; Co-Immunoprecipitations; Collection; Communities; Custom; Development; Diagnostic; Genes; Goals; Human; Immunoblotting; Immunohistochemistry; Immunologics; Island; Libraries; Maps; Measures; Mental Health; Mental disorders; Methods; Molecular; Monoclonal Antibodies; Occupations; Pathology; Pattern; Performance; Phase; Production; Property; Protein Microarray Assay; Proteins; Proteome; Psychiatry; Puerto Rico; Reagent; Recombinant Proteins; Research; Resources; Science; Sensitivity and Specificity; Series; Set protein; Specificity; Staging; Technology; Therapeutic; Training; Transcript; Universities; Validation; Variant; Work; base; chromatin immunoprecipitation; commercialization; cost; design; experience; immunocytochemistry; novel; polyclonal antibody; public health relevance; technology validation; tool; transcription factor

DESCRIPTION (provided by applicant): Antigen-microarray validated monoclonal antibody library for analysis of brain- expressed proteins implicated in human mental disorders. Summary This is a proposal to produce a targeted set of monoclonal antibodies to be commercialized in a variety of ways (see commercialization plan), briefly summarized here as Research (short term), Diagnostics (middle term) and Therapeutics (long term) as a collaborative effort by a startup company in Mayaguez, Puerto Rico, CDI, and the High Throughput Biology Center (HiT) at Johns Hopkins University. The company has developed an efficient pipeline to produce monoclonal antibodies using a complete collection of antigen clones provided by HiT. An antigen array-based protein validation technology developed by the HiT Team forms a critical component of a validation pipeline that allows CDI to rapidly identify antibodies against important and valuable antigens and of very high specificity. The great majority all human genes are expressed in the brain, with many thousands of transcripts expressed in highly cell, region and developmental stage-specific patterns, and recent years have identified many genes as being implicated in the pathology of mental illness. Thus it is critical to have a high-quality set of protein affinity tools to measure, map, and visualize all of these components. Antibodies are the workhorses of protein research because of their use in immunohistochemistry, (co)immunoprecipitation, ChIP on chip, immunoblotting and many other methods. Antibodies vary dramatically, however, in quality; the most commonly used antibodies are polyclonal and thus suffer from significant batch-to-batch variation. Whereas many protein affinity reagents have been described, none has the robust sensitivity and specificity performance of conventional, two-chain antibodies and we focus on streamlining this very well developed technology. We will develop a systematic technology to isolate ultra high specificity renewable monoclonal antibodies, based on exploiting novel antigen microarrays in combination with high throughput production approaches we have developed. Because CDI is located in Puerto Rico, it will be possible to produce large numbers and quantities of mAbs efficiently due to the lower labor costs offshore, while providing excellent training and job opportunities on the island. Our goal in this Phase I application is to produce at least 60 high-grade monoclonal antibodies to transcription factors implicated in human brain development and mental illness. If this approach proves successful, we will expand our effort to produce antibodies to several hundred additional human proteins that are implicated in brain development and/or mental illness in Phase II. PUBLIC HEALTH RELEVANCE: Specific antibodies (or the lack there of) has been a major issue for every colleague we know who has worked in the field of molecular psychiatry. It is a common experience to see an antibody in a catalog advertised as having certain properties or specificity and then finding it to be effectively useless. If the science of molecular psychiatry is to mature, then a series of well validated, reproducible protein affinity reagents is essential. We have developed an approach to rapidly validate antibody specificity. We propose to use this tool to generate a large set of monoclonal antibodies of verified specificity that selectively react with human proteins that are involved in mental illness or brain development. This resource will directly benefit both the mental health research community and the larger biomedical community.

描述（由申请人提供）：抗原微阵列验证的单克隆抗体文库，用于分析与人类精神障碍有关的脑表达蛋白。这是一项生产一组靶向单克隆抗体的提案，将以多种方式商业化（见商业化计划），在此简要总结为研究（短期）、诊断（中期）和治疗（长期），作为波多黎各Mayaguez的一家初创公司、CDI和约翰霍普金斯大学高密度生物学中心（HiT）的合作成果。该公司已经开发出一种高效的管道，使用HiT提供的完整抗原克隆来生产单克隆抗体。HiT团队开发的基于抗原阵列的蛋白质验证技术是验证管道的关键组成部分，使CDI能够快速识别针对重要和有价值抗原的抗体，并且具有非常高的特异性。绝大多数人类基因都在大脑中表达，成千上万的转录本以高度细胞，区域和发育阶段特异性的模式表达，近年来已经鉴定出许多基因与精神疾病的病理学有关。因此，至关重要的是要有一套高质量的蛋白质亲和力工具来测量，映射和可视化所有这些组件。抗体是蛋白质研究的主力，因为它们用于免疫组织化学，（共）免疫沉淀，ChIP芯片，免疫印迹和许多其他方法。然而，抗体的质量差异很大;最常用的抗体是多克隆抗体，因此批次间差异很大。尽管已经描述了许多蛋白质亲和试剂，但没有一种具有常规双链抗体的稳健灵敏度和特异性性能，我们专注于简化这种非常成熟的技术。我们将开发一种系统的技术，以分离超高特异性可再生单克隆抗体，基于开发新的抗原微阵列与我们开发的高通量生产方法相结合。由于CDI位于波多黎各，由于离岸劳动力成本较低，因此可以有效地生产大量mAb，同时在岛上提供良好的培训和就业机会。我们在第一阶段申请的目标是生产至少60种高级别的单克隆抗体，用于治疗与人类大脑发育和精神疾病有关的转录因子。如果这种方法被证明是成功的，我们将扩大我们的努力，以产生数百个额外的人类蛋白质的抗体，这些蛋白质与大脑发育和/或精神疾病有关。 公共卫生关系：特异性抗体（或缺乏特异性抗体）一直是我们认识的每一位在分子精神病学领域工作的同事的主要问题。这是一个共同的经验，看到一个抗体在目录中宣传为具有某些性质或特异性，然后发现它实际上是无用的。如果分子精神病学要走向成熟，那么一系列经过充分验证的、可重复的蛋白质亲和试剂是必不可少的。我们开发了一种快速验证抗体特异性的方法。我们建议使用这个工具来产生一大批具有验证特异性的单克隆抗体，这些抗体选择性地与涉及精神疾病或大脑发育的人类蛋白质反应。这一资源将直接使心理健康研究界和更大的生物医学界受益。