Rapid identification and selection of functional antigen-specific monoclonal antibodies by FcGR-enabled screening on CytoSort Arrays in the CellRaft AIR System

Project SummaryThe production of antibodies using hybridoma screens and 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, sorting and isolating single cells and clonal colonies. By imaging cells on the proprietary CytoSort Array on the CellRaft AIR® System, phenotypes can be characterized in detail and over time, prior to isolating cells and colonies for downstream propagation. During this Phase I program, we will test and develop novel reporter cell lines, software, and cell-based co-culture assays that leverage our CellRaft AIR System as an automated antibody screening platform. Briefly, the CellRaft Technology relies on the CytoSort Array, which contains thousands of microwells, each featuring a releasable plastic 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 AIR System software, cells can be tracked over time for various phenotypes as well as expansion into clonal colonies. The AIR System provides a cost-effective, efficient, and robust platform for screening the production, affinity and functionality of monoclonal antibody producing cells (e.g. hybridomas) prior to sorting so only the most promising candidates need to be harvested. During Phase I, to demonstrate proof-of-concept, we will optimize a novel hybridoma and Jurkat reporter cell line co-culture on the CytoSort Array and evaluate a high throughput workflow for assessing antibody production and functionality. Current technologies offering automated solutions to this challenging workflow are incapable of rivaling the cost savings, throughput, and the detailed phenotypic characterization proposed here.Project NarrativeTargeted monoclonal antibodies have become essential for use as both life science tools and as clinical therapeutics. Although there are currently several methods for producing antibodies, including hybridomas and in vitro immune display libraries, the process is laborious, costly, and inefficient. Screening these antibody libraries requires iterative rounds of affinity screening and cloning without functional validation, which is typically post-screening against a limited number of antibody clones. To improve the efficiency of this process, we propose this proof-of-concept Phase I study using a fully automated method for functionally screening thousands of hybridoma clones on the CytoSort Array and automated CellRaft AIR® System. The CellRaft Technology allows imaging-based sorting and isolation of single cells and clones, thereby allowing sophisticated phenotypic characterization to be incorporated into antibody discovery screening workflows.