A CHO cell-based simultaneous display and secretion platform for accelerating antibody development

Science

The development of novel therapeutic antibodies for the treatment of cancer and infectious diseases continues to face substantial technical and economic challenges. Traditional antibody discovery workflows typically begin with selection in microbial or yeast display systems, followed by a labor-intensive and costly re-engineering process to enable full-length antibody expression in mammalian systems, particularly Chinese hamster ovary (CHO) cells. CHO cells remain the industry standard for biopharmaceutical production due to their ability to support proper protein folding and human-like glycosylation. However, the need to re-clone and re-express antibody candidates in CHO cells introduces inefficiencies, increases timelines, and can compromise antibody functionality. This research addresses a key bottleneck in therapeutic antibody development: the lack of an integrated CHO-based platform that supports both high-throughput screening and small-scale production of functional, full-length antibodies. Such a system would eliminate the disconnect between discovery and production phases, streamlining the development process and reducing overall cost and complexity.

Societal Impact

This research introduces a novel platform designed to streamline therapeutic antibody development by integrating discovery and production within a single Chinese hamster ovary (CHO) cell system. It addresses a critical bottleneck in current biopharmaceutical workflows: the need for re-cloning antibody genes and switching host systems during the transition from screening to production. These steps often introduce inefficiencies, increase the risk of altered antibody properties, and contribute to higher development costs, longer timelines, and elevated failure rates. The CHO-DS platform enables the production of antibody quantities sufficient for a wide range of preclinical functional and developability studies, eliminating the need for separate production processes at this early stage. By eliminating the re-cloning step and need for host cell change, this platform enhances the efficiency of antibody discovery workflow and has the potential to reduce overall development costs and timelines. This would ideally accelerate the path to market for promising new antibody candidates.

Technical Summary

We developed a targeted integration-based CHO cell platform that enables simultaneous antibody display and secretion. Using recombinase-mediated cassette exchange, antibody genes are precisely inserted into a predefined genomic locus, ensuring transcriptional uniformity and high expression levels. We engineered nine-amino-acid FCS variants by incorporating N-terminal P2A residues downstream of the RRKR motif. These variants enabled efficient secretion of antibodies free of 2A remnants while allowing precise modulation of display levels. Notably, the variant with lower display levels enabled discrimination among antibodies with varying binding affinities. We also showed that the platform demonstrated high sensitivity, capable of isolating high-affinity antibody-expressing cells from spike-in populations as rare as 0.0001% from a library.

Figure 1. An overview of the CHO-DS Platform for Antibody Library Screening, Functional Characterization, and Manufacturability Assessment.

References

Ng, J. P., Mariati, M., Bi, J., Chang, M. W., & Yang, Y. (2025). A Targeted Integration-Based CHO Cell Platform for Simultaneous Antibody Display and Secretion. Antibodies, 14(2), 38.