Project Summary The production of antibodies using hybridoma or primary B-cells with in vitro screening technologies represents one of the most industrialized processes in contemporary life science. Products including research reagents, diagnostic tests and biopharmaceuticals rely on the throughput, efficiency and quality of different antibody screening and manufacturing methods. Despite the large-scale and high-quality requirements of these industries, automation of the process for selecting specific antibodies for manufacturing remains an unmet need. The CellRaft Technology represents a novel means of imaging, identifying, and isolating single cells and clonal colonies. By imaging cells on the proprietary CellRaft Array using the CellRaft AIR® System, phenotypes can be characterized in detail and over time, prior to isolating cells and colonies for downstream propagation. During the Phase I program, we tested and developed novel reporter cell lines, software, and cell-based co-culture assays that leveraged our CellRaft AIR System as an automated antibody screening platform. Briefly, the CellRaft Technology relies on the CellRaft Array, which contains thousands of microwells, each featuring a releasable polystyrene floor where cells are seeded and cultured. Cells are phenotypically monitored on the array with the imaging capabilities of the CellRaft AIR System. Using the CellRaft Cytometry analytical software, cells can be tracked over time and analyzed for various phenotypes, including fluorescence intensity, as well as expansion into clonal colonies. The AIR System provides an automated, cost-effective, efficient, and robust platform for screening the production, affinity and functionality of monoclonal antibody producing cells prior to isolation so only the most promising candidates need to be harvested. During Phase II, we will adapt the hybridoma and Jurkat reporter cell line co-culture that was developed in Phase I to be able to screen hundreds of thousands of primary B cells on the CellRaft-HTS Array. We will evaluate a high throughput workflow for assessing production and functionality of novel antibodies against therapeutically relevant antigen targets. Current technologies offering automated solutions to this challenging workflow are incapable of rivaling the cost savings, throughput, and the detailed phenotypic and functional characterization proposed here.