Production and Purification of High-Titer Newcastle Disease Virus for Use in Preclinical Mouse Models of Cancer

• Published in: Molecular therapy. Methods & clinical development Vol. 9; pp. 181 - 191
• Main Authors: Santry, Lisa A, McAusland, Thomas M, Susta, Leonardo, Wood, Geoffrey A, Major, Pierre P, Petrik, Jim J, Bridle, Byram W, Wootton, Sarah K
• Format: Journal Article
• Language: English
• Published: United States Elsevier Limited 15-06-2018; American Society of Gene & Cell Therapy; Elsevier
• Subjects: Allantoic fluid; Animal models; Cancer; Cell culture; Chromatography; Clinical trials; Contaminants; Eggs; Genomes; Humidity; Immunogenicity; Impurities; Intravenous administration; Laboratories; Medical research; Methods; Molecular weight; Newcastle disease; Oncolysis; Plasmids; Polyethylene glycol; RNA polymerase; RNA viruses; Viruses; Canada; intravenous; Newcastle disease virus; preclinical grade; oncolytic virus; tangential flow filtration; allantoic fluid
• ISSN: 2329-0501; 2329-0501
• DOI: 10.1016/j.omtm.2017.10.004

Newcastle disease virus (NDV) is a single-stranded, negative-sense RNA virus in the
family. Although primarily an avian pathogen, NDV is a potent oncolytic virus that has been shown to be safe and effective in a variety of preclinical cancer models and human clinical trials. To produce virus for oncolytic trials, NDV is commonly amplified in embryonated chicken eggs and purified from the allantoic fluid. Conventional methods for purifying virus from allantoic fluid often result in relatively low-titer preparations containing high levels of impurities, including immunogenic chicken host cell proteins from allantoic fluid. However, large quantities of virus need to be delivered intravenously to administer oncolytic NDV systemically to mice. This route of administration requires virus preparations that are both highly concentrated (to enable delivery of small volumes) and highly pure (to limit toxic effects from contaminants). Given the accumulation of promising preclinical and clinical data demonstrating the efficacy of NDV as an oncolytic agent, strategies for increasing the titer and purity of NDV preparations are sorely needed to allow for effective intravenous administration in mice. Here, we describe an optimized protocol for the rescue, production, and purification of high-titer
-grade NDV for preclinical studies in mouse models.