Monospecific monoclonal antibodies against human transcription factors

描述（由申请人提供）：我们建议创建一个生产中心，该中心拥有高效且具有成本效益的非特异性单克隆抗体（mMAb）管线，将生产识别1500种人转录因子的蛋白捕获试剂。该生产中心利用人类蛋白质组微阵列作为一种高成本效益的特异性分析工具，并作为一个全面的生化表征和验证管道，包括免疫细胞化学，免疫印迹和免疫沉淀性能评估和芯片熟练度分析的关键步骤。此外，我们将开发一种新的方法，将先进的光学技术，斜入射反射率差，与蛋白质微阵列（OIRD-PM）相结合，这将使抗体亲和力测量的成本极低。我们还将开发一个细胞微阵列平台，该平台可以提供一个极低成本的平台，用于评估免疫组织化学的性能，这是抗体的高度挑战性应用。CDI Inc.的抗体生产平台位于波多黎各，低劳动力成本和训练有素的劳动力联合收割机相结合，生产世界一流的亲和试剂便宜和可扩展。针对全细胞和细胞组分产生的抗体的质量，随后是蛋白质组微阵列解卷积和验证，产生最高质量的mAb。一个创新的实习计划将带来来自CDI的顶级技术表演者在我们的中心在约翰霍普金斯大学医学院先进的traipsing。我们强调在我们的生产工作中实现高吞吐量和可扩展性的优势，并建议开发工艺改进，从而在获奖期间提高成本和质量。我们将保持一定程度的科学灵活性，以开发生产亲和试剂的方法，这些亲和试剂针对在生产过程中鉴定的不太容易处理的转录因子蛋白质靶点。最后，我们提出了一个可行的分配计划，利用现有的开发性杂交瘤单克隆库，确保广泛的可用性和广泛的可及性，以非常低的成本和最小的限制，为未来使用的试剂，符合这一资助计划的目标

DESCRIPTION (provided by applicant): We propose to create a production center with a highly efficient and cost-effective nonspecific Monoclonal antibody (mMAb) pipeline that will produce such Protein Capture reagents recognizing 1500 human Transcription Factors. This production center exploits a Human Proteome Microarray as a highly cost effective specificity profiling tool and as a critical step in a thorough biochemical characterization and validation pipeline that includes immunocytochemistry, assessment of performance in immunoblotting and immunprecipitafion and analyses of Chip-proficiency. Additionally we will develop a novel methodology marrying an advanced optical technique, Oblique Incidence Reflectivity Difference, with Protein Microarrays (OIRD-PM) that will enable extremely low cost quantification of antibody affinity measurement. We will also develop a Cell Microarray platform that could provide an extremely low cost platform for assessing performance in immunohistochemistry, highly challenging application for antibodies. Our antibody production platform at CDI Inc. is located in Puerto Rico, where low labor costs and a highly trained workforce combine to produce world class affinity reagents cheaply and scalable. The quality of antibodies raised against whole cells and cell fractions, followed by Proteome Microarray deconvolufion and validation, produces mAbs of the highest quality. An innovative internship program will bring top technical performers from CDI for advanced traipsing in our Center at Johns Hopkins University School of Medicine. We emphasize the advantages of high throughput and scalability to be achieved within our production effort, and propose the development of process improvements, thereby improving cost and quality over the duration of the award. We will maintain a degree of scientific flexibility to develop approaches to producing affinity reagents to less tractable transcription factor protein targets as they are identified in the course of production. Finally we propose a viable distribution plan utilizing the existing Developmental Hybridoma Monoclonal Bank, which ensures broad availability and wide accessibility for future use of the reagents at very low cost and with minimal constraints, consistent with the goals of this funding initiative