title: Synthetic generation of influenza vaccine viruses for rapid response to pandemics.
abstract: During the 2009 H1N1 influenza pandemic, vaccines for the virus became available in large quantities only after human infections peaked. To accelerate vaccine availability for future pandemics, we developed a synthetic approach that very rapidly generated vaccine viruses from sequence data. Beginning with hemagglutinin (HA) and neuraminidase (NA) gene sequences, we combined an enzymatic, cell-free gene assembly technique with enzymatic error correction to allow rapid, accurate gene synthesis. We then used these synthetic HA and NA genes to transfect Madin-Darby canine kidney (MDCK) cells that were qualified for vaccine manufacture with viral RNA expression constructs encoding HA and NA and plasmid DNAs encoding viral backbone genes. Viruses for use in vaccines were rescued from these MDCK cells. We performed this rescue with improved vaccine virus backbones, increasing the yield of the essential vaccine antigen, HA. Generation of synthetic vaccine seeds, together with more efficient vaccine release assays, would accelerate responses to influenza pandemics through a system of instantaneous electronic data exchange followed by real-time, geographically dispersed vaccine production.
authors.affiliation: Novartis Vaccines and Diagnostics, Cambridge, MA 02139, USA. philip.dormitzer@novartis.com
authors.forename: Philip R
authors.initials: PR
authors.lastname: Dormitzer
authors.forename: Pirada
authors.initials: P
authors.lastname: Suphaphiphat
authors.forename: Daniel G
authors.initials: DG
authors.lastname: Gibson
authors.forename: David E
authors.initials: DE
authors.lastname: Wentworth
authors.forename: Timothy B
authors.initials: TB
authors.lastname: Stockwell
authors.forename: Mikkel A
authors.initials: MA
authors.lastname: Algire
authors.forename: Nina
authors.initials: N
authors.lastname: Alperovich
authors.forename: Mario
authors.initials: M
authors.lastname: Barro
authors.forename: David M
authors.initials: DM
authors.lastname: Brown
authors.forename: Stewart
authors.initials: S
authors.lastname: Craig
authors.forename: Brian M
authors.initials: BM
authors.lastname: Dattilo
authors.forename: Evgeniya A
authors.initials: EA
authors.lastname: Denisova
authors.forename: Ivna
authors.initials: I
authors.lastname: De Souza
authors.forename: Markus
authors.initials: M
authors.lastname: Eickmann
authors.forename: Vivien G
authors.initials: VG
authors.lastname: Dugan
authors.forename: Annette
authors.initials: A
authors.lastname: Ferrari
authors.forename: Raul C
authors.initials: RC
authors.lastname: Gomila
authors.forename: Liqun
authors.initials: L
authors.lastname: Han
authors.forename: Casey
authors.initials: C
authors.lastname: Judge
authors.forename: Sarthak
authors.initials: S
authors.lastname: Mane
authors.forename: Mikhail
authors.initials: M
authors.lastname: Matrosovich
authors.forename: Chuck
authors.initials: C
authors.lastname: Merryman
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Palladino
authors.forename: Gene A
authors.initials: GA
authors.lastname: Palmer
authors.forename: Terika
authors.initials: T
authors.lastname: Spencer
authors.forename: Thomas
authors.initials: T
authors.lastname: Strecker
authors.forename: Heidi
authors.initials: H
authors.lastname: Trusheim
authors.forename: Jennifer
authors.initials: J
authors.lastname: Uhlendorff
authors.forename: Yingxia
authors.initials: Y
authors.lastname: Wen
authors.forename: Anthony C
authors.initials: AC
authors.lastname: Yee
authors.forename: Jayshree
authors.initials: J
authors.lastname: Zaveri
authors.forename: Bin
authors.initials: B
authors.lastname: Zhou
authors.forename: Stephan
authors.initials: S
authors.lastname: Becker
authors.forename: Armen
authors.initials: A
authors.lastname: Donabedian
authors.forename: Peter W
authors.initials: PW
authors.lastname: Mason
authors.forename: John I
authors.initials: JI
authors.lastname: Glass
authors.forename: Rino
authors.initials: R
authors.lastname: Rappuoli
authors.forename: J Craig
authors.initials: JC
authors.lastname: Venter
mesh_terms: D000818:Animals; D002460:Cell Line; D003198:Computer Simulation; D004285:Dogs; D005813:Genes, Synthetic; D019267:Hemagglutinin Glycoproteins, Influenza Virus / Q000235:genetics; D006801:Humans; D064766:Influenza A Virus, H7N9 Subtype / Q000276:immunology; D009980:Influenza A virus / Q000276:immunology*; D007252:Influenza Vaccines / Q000276:immunology*; D007251:Influenza, Human / Q000276:immunology* / Q000517:prevention & control* / Q000821:virology; D061985:Madin Darby Canine Kidney Cells; D009439:Neuraminidase / Q000235:genetics; D058873:Pandemics / Q000517:prevention & control*; D016865:Reassortant Viruses / Q000276:immunology; D015203:Reproducibility of Results; D014614:Vaccines, Synthetic / Q000276:immunology*; D019562:Viral Load
chemical_list: D019267:Hemagglutinin Glycoproteins, Influenza Virus; D007252:Influenza Vaccines; D014614:Vaccines, Synthetic; D009439:Neuraminidase