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

描述(由申请人提供)：抗原微阵列验证的单抗文库，用于分析与人类精神障碍有关的脑表达蛋白。摘要这是一项以多种方式(参见商业化计划)生产一组目标单抗以商业化的建议，在这里简要总结为研究(短期)、诊断(中期)和治疗(长期)，这是由波多黎各马亚圭兹的一家初创公司、CDI和约翰·霍普金斯大学的高通量生物学中心(HIT)共同努力的结果。该公司已经开发了一条高效的管道，使用HIT提供的一整套抗原克隆来生产单克隆抗体。HIT团队开发的一种基于抗原阵列的蛋白质验证技术形成了验证管道的关键组件，使CDI能够快速识别针对重要和有价值的抗原的抗体，并具有非常高的特异性。绝大多数人类基因在大脑中表达，数以千计的转录本以高度细胞、区域和发育阶段的模式表达，近年来发现许多基因与精神疾病的病理有关。因此，拥有一套高质量的蛋白质亲和力工具来测量、绘制和可视化所有这些成分是至关重要的。抗体是蛋白质研究的主力，因为它们用于免疫组织化学、免疫共沉淀、芯片、免疫印迹和许多其他方法。然而，抗体在质量上差异很大；最常用的抗体是多克隆的，因此在批次之间存在显著的差异。虽然已经描述了许多蛋白质亲和试剂，但没有一种具有传统双链抗体的强大敏感性和特异性，我们专注于简化这项非常成熟的技术。我们将在开发新的抗原微阵列的基础上，结合我们开发的高通量生产方法，开发一种分离超高特异性可再生单抗的系统技术。由于CDI位于波多黎各，由于离岸劳动力成本较低，将有可能高效地生产大量和大量的单抗，同时在岛上提供极好的培训和就业机会。我们在这一阶段的应用中的目标是生产至少60个针对与人脑发育和精神疾病有关的转录因子的高级单抗。如果这种方法被证明是成功的，我们将扩大我们的努力，在第二阶段生产与大脑发育和/或精神疾病有关的数百种额外人类蛋白的抗体。 公共卫生相关性：对于我们认识的每一位在分子精神病学领域工作过的同事来说，特定的抗体(或缺乏抗体)一直是一个主要问题。在目录中看到一种抗体被宣传为具有某些特性或特异性，然后发现它实际上是无用的，这是一种常见的经验。如果分子精神病学的科学要成熟，那么一系列经过充分验证的、可重复性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。我们建议使用这一工具来产生一大组经过验证的特异性单抗，这些单抗可以选择性地与涉及精神疾病或大脑发育的人类蛋白发生反应。这一资源将直接惠及心理健康研究界和更大的生物医学界。

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.