The production of antibodies using hybridoma or primary B-cells with in vitro screening technologiesrepresents one of the most industrialized processes in contemporary life science. Products including researchreagents, diagnostic tests and biopharmaceuticals rely on the throughput, efficiency and quality of differentantibody screening and manufacturing methods. Despite the large-scale and high-quality requirements of theseindustries, 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 clonalcolonies. By imaging cells on the proprietary CellRaft Array using the CellRaft AIR® System, phenotypes canbe characterized in detail and over time, prior to isolating cells and colonies for downstream propagation. Duringthe Phase I program, we tested and developed novel reporter cell lines, software, and cell-based co-cultureassays that leveraged our CellRaft AIR System as an automated antibody screening platform. Briefly, theCellRaft Technology relies on the CellRaft Array, which contains thousands of microwells, each featuring areleasable polystyrene floor where cells are seeded and cultured. Cells are phenotypically monitored on thearray 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 asexpansion into clonal colonies. The AIR System provides an automated, cost-effective, efficient, and robustplatform for screening the production, affinity and functionality of monoclonal antibody producing cells prior toisolation so only the most promising candidates need to be harvested. During Phase II, we will adapt thehybridoma and Jurkat reporter cell line co-culture that was developed in Phase I to be able to screen hundredsof thousands of primary B cells on the CellRaft-HTS Array. We will evaluate a high throughput workflow forassessing 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 costsavings, throughput, and the detailed phenotypic and functional characterization proposed here.
Public Health Relevance Statement: Project Narrative
Targeted 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 Phase II study using a fully automated method for functionally screening hundreds of thousands of primary
B-cells from immunized animals on the high throughput CellRaft-HTS 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.
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