TY - JOUR
T1 - Stability of retroviral pseudotypes carrying the hemagglutinin of avian influenza viruses under various storage conditions
AU - Delgadillo-Gutiérrez, Karen
AU - Ribas-Aparicio, Rosa María
AU - Jiménez-Alberto, Alicia
AU - Aparicio-Ozores, Gerardo
AU - Castelán-Vega, Juan A.
N1 - Publisher Copyright:
© 2018 Elsevier B.V.
PY - 2019/1
Y1 - 2019/1
N2 - 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 10 6 RLU/mL, which decreased to 10 5 –10 6 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 log 10 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.
AB - 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 10 6 RLU/mL, which decreased to 10 5 –10 6 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 log 10 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.
KW - Avian influenza
KW - Hemagglutinin
KW - Neutralization assay
KW - Retroviral pseudotypes
UR - http://www.scopus.com/inward/record.url?scp=85055744672&partnerID=8YFLogxK
U2 - 10.1016/j.jviromet.2018.10.013
DO - 10.1016/j.jviromet.2018.10.013
M3 - Artículo
C2 - 30347199
SN - 0166-0934
VL - 263
SP - 44
EP - 49
JO - Journal of Virological Methods
JF - Journal of Virological Methods
ER -