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
(same as original) A consistent set of renewable protein Capture Reagents will facilitate NIH research in all areas relevant to human health as a basic tool used by virtually all NIH funded scientists. Monoclonal antibodies will also provide the diagnostic tools and therapeutic medicines of the future.
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