Mammalian display for high-throughput antibody screening

Abstract

Antibodies have great potential for their use in immune therapies. Developing potent therapeutic monoclonal antibodies for the treatment of cancer and infectious diseases is a critical task. Although there have been advancements in display technologies to simplify the process, the major bottleneck in most existing methods is that they depend on examining one antigen against a pool of antibodies, significantly decreasing the overall throughput. In this study, we report a high-throughput assay for screening of antibody library against antigen library. It leverages a mammalian display system to express proteins of interest on the surface of cells to enrich specific cell doublets, while eliminating the formation of unspecific binding events. The pairing information is retrieved through a fusion PCR post droplet encapsulation of the cell-doublets. As a proof of concept, cells expressing a nanobody against GFP on their membrane were successfully enriched in the form of cell doublets along with GFP expressing cells utilizing this binding assay. This was demonstrated even at lower ratios of protein-specific cells in the mixture. These results suggest that different protein display libraries can be deployed to discover new protein interactors using this approach. Using this technology, we offer a strategy that could allow for high throughput screening of antibodies which can be beneficial for immunological research.