title: Novel formulations enhance the thermal stability of live-attenuated flavivirus vaccines.
abstract: Thermal stability is important for the manufacture, distribution and administration of vaccines, especially in tropical developing countries, where particularly adverse field conditions exist. Current live-attenuated flavivirus vaccines exhibit relatively poor liquid stability in clinical settings, and clinicians are instructed to discard the yellow fever vaccine 1h after reconstitution. We have identified novel combinations of excipients that greatly enhance the thermal stability of live-attenuated DEN-2 PDK-53-based flavivirus vaccine candidates. Liquid formulations comprising a sugar, albumin and a pluronic polymer minimized the loss of flavivirus infectious titer to less than 0.5 log(10)pfu after storage for at least 8h at 37°C, 7 days at room temperature or at least 11 weeks at 4°C. Additionally, these formulations prevented reduction of viral infectivity after two freeze-thaw cycles of virus. Formulated candidate vaccines were readily lyophilized and reconstituted with minimal loss of viral titers. In mice, the formulations were safe and did not hinder the ability of the vaccine virus to generate a potent, protective immune response. These formulations provided significantly greater liquid-phase stability than has been reported previously for other flavivirus vaccine formulations. The enhanced thermal stability provided by the formulations described here will facilitate the effective distribution of flavivirus vaccines worldwide.
authors.affiliation: Inviragen Inc., Fort Collins, Colorado 80525,United States.
authors.forename: O'Neil
authors.initials: O
authors.lastname: Wiggan
authors.forename: Jill A
authors.initials: JA
authors.lastname: Livengood
authors.forename: Shawn J
authors.initials: SJ
authors.lastname: Silengo
authors.forename: Richard M
authors.initials: RM
authors.lastname: Kinney
authors.forename: Jorge E
authors.initials: JE
authors.lastname: Osorio
authors.forename: Claire Y-H
authors.initials: CY
authors.lastname: Huang
authors.forename: Dan T
authors.initials: DT
authors.lastname: Stinchcomb
mesh_terms: D000418:Albumins; D000818:Animals; D002626:Chemistry, Pharmaceutical; D002522:Chlorocebus aethiops; D053059:Dengue Vaccines / Q000008:administration & dosage / Q000276:immunology; D003716:Dengue Virus / Q000276:immunology; D004355:Drug Stability; D004356:Drug Storage; D051379:Mice; D009500:Neutralization Tests; D011108:Polymers; D055550:Protein Stability; D013696:Temperature; D014613:Vaccines, Attenuated / Q000008:administration & dosage / Q000276:immunology; D014709:Vero Cells; D014765:Viral Vaccines / Q000008:administration & dosage / Q000276:immunology; D014902:West Nile virus / Q000276:immunology*; D022341:Yellow Fever Vaccine / Q000008:administration & dosage / Q000276:immunology; D015005:Yellow fever virus / Q000276:immunology
chemical_list: D000418:Albumins; D053059:Dengue Vaccines; D011108:Polymers; D014613:Vaccines, Attenuated; D014765:Viral Vaccines; D022341:Yellow Fever Vaccine