Journal article

An Influenza A/H1N1/2009 hemagglutinin vaccine produced in Escherichia coli

Aguilar-Yáñez J, Portillo-Lara R, Mendoza-Ochoa G, García-Echauri S, López-Pacheco F, Bulnes-Abundis D, Salgado-Gallegos J, Lara-Mayorga I, Webb-Vargas Y, León-Angel F, Rivero-Aranda R, Oropeza-Almazán Y, Ruiz-Palacios G, Zertuche-Guerra M, DuBois R, White S, Schultz-Cherry S, Russell C, Alvarez M ...see all

PLoS ONE, vol. 5, issue 7 (2010)

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BACKGROUND: The A/H1N1/2009 influenza pandemic made evident the need for faster and higher-yield methods for the production of influenza vaccines. Platforms based on virus culture in mammalian or insect cells are currently under investigation. Alternatively, expression of fragments of the hemagglutinin (HA) protein in prokaryotic systems can potentially be the most efficacious strategy for the manufacture of large quantities of influenza vaccine in a short period of time. Despite experimental evidence on the immunogenic potential of HA protein constructs expressed in bacteria, it is still generally accepted that glycosylation should be a requirement for vaccine efficacy.

METHODOLOGY/PRINCIPAL FINDINGS: We expressed the globular HA receptor binding domain, referred to here as HA(63-286)-RBD, of the influenza A/H1N1/2009 virus in Escherichia coli using a simple, robust and scalable process. The recombinant protein was refolded and purified from the insoluble fraction of the cellular lysate as a single species. Recombinant HA(63-286)-RBD appears to be properly folded, as shown by analytical ultracentrifugation and bio-recognition assays. It binds specifically to serum antibodies from influenza A/H1N1/2009 patients and was found to be immunogenic, to be capable of triggering the production of neutralizing antibodies, and to have protective activity in the ferret model.

CONCLUSIONS/SIGNIFICANCE: Projections based on our production/purification data indicate that this strategy could yield up to half a billion doses of vaccine per month in a medium-scale pharmaceutical production facility equipped for bacterial culture. Also, our findings demonstrate that glycosylation is not a mandatory requirement for influenza vaccine efficacy.

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  • José M. Aguilar-Yáñez

  • Roberto Portillo-Lara

  • Gonzalo I. Mendoza-Ochoa

  • Sergio A. García-Echauri

  • Felipe López-Pacheco

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