Abstract
Influenza viruses infect millions of people each year, resulting in significant morbidity and mortality in the human population. Therefore, generation of a universal influenza virus vaccine is an urgent need and would greatly benefit public health. Recombinant protein technology is an established vaccine platform and has resulted in several commercially available vaccines. Herein, we describe the approach for developing stable transfected human cell lines for the expression of recombinant influenza virus hemagglutinin (HA) and recombinant influenza virus neuraminidase (NA) proteins for the purpose of in vitro and in vivo vaccine development. HA and NA are the main surface glycoproteins on influenza virions and the major antibody targets. The benefits for using recombinant proteins for in vitro and in vivo assays include the ease of use, high level of purity and the ability to scale-up production. This work provides guidelines on how to produce and purify recombinant proteins produced in mammalian cell lines through either transient transfection or generation of stable cell lines from plasmid creation through the isolation step via Immobilized Metal Affinity Chromatography (IMAC). Collectively, the establishment of this pipeline has facilitated large-scale production of recombinant HA and NA proteins to high purity and with consistent yields, including glycosylation patterns that are very similar to proteins produced in a human host.
Highlights
Recombinant proteins are an accepted vaccine platform and are components in a range of commercially available vaccines including the hepatitis B (HBV) and human papilloma virus (HPV)vaccines [1,2,3]
HA and NA are the major glycoproteins expressed on the surface of influenza viral particles and the major targets of the humoral response [4,5]
Vaccines 2020, 8, 462 strategies based on administration of recombinant proteins enable focusing of the immune response toward antigens associated with protective immunity [6]
Summary
Recombinant proteins are an accepted vaccine platform and are components in a range of commercially available vaccines including the hepatitis B (HBV) and human papilloma virus (HPV)vaccines [1,2,3]. Usage of recombinant proteins as immunogens circumvents issues associated with alternative influenza vaccine production systems, such as the occurrence of adaptive mutations using embryonated hen eggs for virus growth [7]. For this reason, current under-development generation influenza vaccines mostly rely on a recombinant protein-based platform [7,8]. The here described protein production platform can be considered safer and more affordable as compared, for example, to transducing systems such as those relying on recombinant lentivirus [9]. Lentiviral-based systems often require a higher biosafety capacity [10]
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