Stability of retroviral pseudotypes carrying the hemagglutinin of avian influenza viruses under various storage conditions

J Virol Methods. 2019 Jan:263:44-49. doi: 10.1016/j.jviromet.2018.10.013. Epub 2018 Oct 19.

Abstract

Retroviral pseudotypes are broadly used as safe instruments to mimic the structure and surface of highly pathogenic viruses. They have been employed for the discovery of new drugs, as diagnostic tools in vaccine studies, and part of serological assays. Because of their widespread use in research and their potential as tools for quality control, it is important to know their shelf life, stability, and best storage conditions. In this study, we produced pseudotypes carrying the lacZ reporter gene and the hemagglutinin (HA) of avian influenza virus subtypes H5 and H7 to investigate their stability under various storage conditions. We produced pseudotypes with titers of approximately 106 RLU/mL, which decreased to 105-106 RLU/mL after short-term storage at 4 °C (up to 4 weeks). Stability was maintained after long-term storage at -20 °C (up to 12 months), even under storage variations such as freeze-thaw cycles. We conclude that, although the titers decreased by 1 log10 under the different storage conditions, the remaining titers can be readily applicable in many techniques, such as neutralization assays. These findings show that large quantities of retroviral pseudotypes can be safely stored for short- or long-term use, allowing standardization and reduced variation in assays involving retroviral pseudotypes.

Keywords: Avian influenza; Hemagglutinin; Neutralization assay; Retroviral pseudotypes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cold Temperature
  • Defective Viruses / genetics
  • Defective Viruses / physiology
  • Genes, Reporter / genetics
  • Genetic Vectors / genetics
  • Genetic Vectors / physiology
  • HEK293 Cells
  • Hemagglutinin Glycoproteins, Influenza Virus / genetics*
  • Humans
  • Influenza A virus / genetics*
  • Lentivirus / genetics
  • Lentivirus / physiology*

Substances

  • Hemagglutinin Glycoproteins, Influenza Virus