Effect of chemical stabilizers on the thermostability and infectivity of a representative panel of freeze dried viruses

PLoS One. 2015 Apr 29;10(4):e0118963. doi: 10.1371/journal.pone.0118963. eCollection 2015.

Abstract

As a partner of the European Virus Archive (EVA) FP7 project, our laboratory maintains a large collection of freeze-dried viruses. The distribution of these viruses to academic researchers, public health organizations and industry is one major aim of the EVA consortium. It is known that lyophilization requires appropriate stabilizers to prevent inactivation of the virus. However, few studies have investigated the influence of different stabilizers and lyophilization protocols on the thermostability of different viruses. In order to identify optimal lyophilization conditions that will deliver maximum retention of viral infectivity titre, different stabilizer formulations containing trehalose, sorbitol, sucrose or foetal bovine serum were evaluated for their efficacy in stabilizing a representative panel of freeze dried viruses at different storage temperatures (-20°C, +4°C and +20°C) for one week, the two latter mimicking suboptimal shipping conditions. The Tissue Culture Infectious Dose 50% (TCID50) assay was used to compare the titres of infectious virus. The results obtained using four relevant and model viruses (enveloped/non enveloped RNA/DNA viruses) still serve to improve the freeze drying conditions needed for the development and the distribution of a large virus collection.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line
  • Excipients / pharmacology*
  • Freeze Drying / methods*
  • Humans
  • RNA Viruses / drug effects
  • RNA Viruses / pathogenicity*
  • Temperature*

Substances

  • Excipients

Grants and funding

This work was supported in part by (i) the European Virus Archive (European Union 7th Framework Programme under grant agreement no. 228292)(http://www.european-virus-archive.com/), (ii) the EDENext FP7- n°261504 EU project and this paper is catalogued by the EDENext Steering Committee (http://www.edenext.eu), (iii) European Network for Diagnostics of "Imported" Viral Diseases (ENIVD) funded from the European Centre for Disease Prevention and Control (http://www.enivd.de/index.htm). The work of RNC was done under the frame of EurNegVec COST Action TD1303. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.