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Flow Electroporation for Vaccine Development and Production: From Subunit Vaccines to Ex Vivo Immunotherapy

August 15, 2014

MaxCyte flow electroporation is a universal, clinically validated transient transfection platform for rapid, high-quality cell transfection. Flow electroporation enables (co)transfection of a wide range of cells with DNA, RNA, proteins, or cell lysates using single-use processing assemblies for cGMP, “plug-and-play” manufacturing of recombinant proteins and vaccines. This technology combines superior performance, broad applicability, and ease of use with the capacity to transfect up to 2E11 cells in under 30 minutes, creating a fully scalable, highly reproducible, and regulatory-compliant transfection method. No other single expression system—stable cells, baculoviruses, lipid- or chemical-based transient transfection—offers the ability to develop and manufacture the full complement of next-generation vaccines, including therapeutic antibodies and antibody-like molecules, subunit vaccines, virus-like particles (VLPs), virus-like replicon particles (VRPs), and viral vectors, as well as ex vivo cellular immunotherapies such as CAR engineering and dendritic cell loading. This technical note reviews MaxCyte flow electroporation and its applications in the development and production of vaccines and cellular immunotherapies.

 

Vaccination has proven to be a successful and cost-effective public health intervention. It has eradicated many illnesses (e.g., smallpox) and provided drastic reductions in others (e.g., polio). Yet, vaccine development and production is often a lengthy and costly process, as exemplified by the 2009-2010 swine H1N1 flu outbreak in which egg-based manufacturing was unable to provide sufficiently large numbers of vaccine doses in a timely manner (1). In response to the need for rapid and reliable vaccine production, researchers have looked to recombinant technologies to develop innovative types of vaccines and new cell culture-based means of production that offer shorter lead times and greater production flexibility while maintaining vaccine safety (2-4). Newer, engineered vaccine modalities range from therapeutic antibodies, subunit vaccines, VLPs, and VRPs to virus-mediated gene therapy and ex vivo cellular immunotherapies such as dendritic cell loading and chimeric antigen receptor T-cell targeting.

 

Each of these engineered vaccine types—whether a simple therapeutic protein or complex modification of patient cells—requires the introduction of recombinant nucleic acids into cell lines or ex vivo patient-isolated cells via transfection and/or the creation of stable cell lines. While the downstream processing is vaccine-type dependent, a single unifying platform for upstream cell transfection could significantly reduce development timelines and production costs, if it fulfills the overarching needs for safety, flexibility, and scalability.

 

For more than two decades, stable cell lines have been the standard for biotherapeutic protein production; however, their creation is a costly, time-consuming, and labor-intensive process and is not feasible for all vaccine applications. In response, researchers have looked to transient gene expression (TGE) as a means of more cost-effective protein production, particularly during early development and preclinical stages (5, 6). While TGE generally offers a means of rapidly expressing proteins, not all transient expression methods fulfill the necessary requirements for broad use throughout vaccine development and production.

 

Transient transfection technologies have evolved from simple chemical methods such as PEI to sophisticated methodologies such as lipid-based reagents, baculovirus expression systems, and electroporation. MaxCyte’s proprietary flow electroporation technology is a universal, regulatory-compliant transient transfection platform that provides a practical solution to the time, labor, and cost challenges of developing stable cell lines and baculovirus-based expression while overcoming the flexibility and scalability limitations associated with other transient transfection methods.

 

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