The biopharmaceutical industry needs efficient, high-yield expression systems for recombinant proteins and viral vectors, as current HEK293-based processes suffer from complex plasmid designs, cloning challenges with repetitive EBNA1 sequences, and suboptimal productivity, slowing development of serological tests and gene therapy products.
This project will design and test optimized plasmid constructs with simplified EBNA1 sequences and high-performing promoters (CMV, CAG, SV40) to overcome cloning complexity and improve recombinant protein expression in HEK293 cells, enabling up to 2.1× higher titers and faster development of antigens and gene therapy components while maintaining quality and compliance.
Impact on the development of the principal discipline(s) of the project. Developed an improved method for expression of coronavirus recombinant antigens using HEK293 cells. The work has been published in the Open Access scientific literature.
Impact on the development of human resources. Two PhD students successfully completed their research doctorates and had an additional peer-reviewed publication as a result.
Impact on teaching and educational experiences. Two PhD students successfully completed their research doctorates and had an additional peer-reviewed publication as a result.
Impact on physical, institutional, and information resources that form infrastructure: not applicable
Impact on society beyond science and technology. this research knowledge may result in improved expression of recombinant antigens using HEK293 cells thereby accelerating the time between identification of a coronavirus public health threat and response.
By implementing optimized plasmid designs with simplified EBNA1 sequences and high-performing promoters (CMV, CAG, SV40), as demonstrated in this project, an organization will reduce recombinant antigen production costs by improving titers up to 2.1× and eliminating cloning complexity associated with repetitive EBNA1 regions, enabling faster development of serological test antigens and gene therapy components.
Green, E.A., Hamaker, N.K., & Lee, K.H., (2023) Comparison of vector elements and process conditions in transient and stable suspension HEK293 platforms using SARS-CoV-2 receptor binding domain as a model protein, BMC Biotechnology 23, 7. https://doi.org/10.1186/s12896-023-00777-7
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University of Delaware
