Lyophilisation of influenza, rabies and Marburg lentiviral pseudotype viruses for the development and distribution of a neutralisation-assay based diagnostic kit

Mather, Stuart, Wright, Edward, Scott, Simon D., Temperton, Nigel J. (2014) Lyophilisation of influenza, rabies and Marburg lentiviral pseudotype viruses for the development and distribution of a neutralisation-assay based diagnostic kit. Journal of Virological Methods, 210 . pp. 51-58. ISSN 0166-0934. (doi:10.1016/j.jviromet.2014.09.021)

PDF - Author's Accepted Manuscript
Download (826kB) Preview
[img]
Preview
Official URL
http://dx.doi.org/10.1016/j.jviromet.2014.09.021

Abstract

Pseudotype viruses (PVs) are chimeric, replication-deficient virions that mimic wild-type virus entry mechanisms and can be safely employed in neutralisation assays, bypassing the need for high biosafety requirements and performing comparably to established serological assays. However, PV supernatant necessitates -80°C long-term storage and cold-chain maintenance during transport, which limits the scope of dissemination and application throughout resource-limited laboratories. We therefore investigated the effects of lyophilisation on influenza, rabies and Marburg PV stability, with a view to developing a pseudotype virus neutralisation assay (PVNA) based kit suitable for affordable global distribution. Infectivity of each PV was calculated after lyophilisation and immediate reconstitution, as well as subsequent to incubation of freeze-dried pellets at varying temperatures, humidities and timepoints. Integrity of glycoprotein structure following treatment was also assessed by employing lyophilised PVs in downstream PVNAs. In the presence of 0.5M sucrose-PBS cryoprotectant, each freeze-dried pseudotype was stably stored for 4 weeks at up to 37°C and could be neutralised to the same potency as unlyophilised PVs when employed in PVNAs. These results confirm the viability of a freeze-dried PVNA-based kit, which could significantly facilitate low-cost serology for a wide portfolio of emerging infectious viruses.

Item Type: Article
DOI/Identification number: 10.1016/j.jviromet.2014.09.021
Uncontrolled keywords: Retroviral pseudotype viruses; Lyophilisation; Neutralising antibodies; Serological assays
Subjects: Q Science > QR Microbiology > QR355 Virology
Divisions: Faculties > Sciences > Medway School of Pharmacy
Depositing User: Nigel Temperton
Date Deposited: 24 Sep 2014 13:00 UTC
Last Modified: 29 May 2019 13:06 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/43055 (The current URI for this page, for reference purposes)
Scott, Simon D.: https://orcid.org/0000-0002-8290-0461
Temperton, Nigel J.: https://orcid.org/0000-0002-7978-3815
  • Depositors only (login required):

Downloads

Downloads per month over past year