Rhabdoviridae virus preparations
AssigneeMayo Foundation for Medical Education and Research
InventorsMark J. Federspiel, Troy R. Wegman, Kirsten K. Langfield, Henry J. Walker, Sharon A. Stephan
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Abstract
This document involves methods and materials related to obtaining Rhabdoviridae virus preparations. For example, methods and materials for obtaining large volume, high titer, high purity preparations of Rhabdoviridae viruses (e.g., VSV) with low or non-existent levels of contaminants are provided.
Claims21
A method for producing Rhabdoviridae viruses at titers of at least 108 TCID50 per mL directly in cell culture supernatants, said method comprising:
- (a) growing producer cells in serum-free medium to a cell density,
- (b) infecting said cells with a Rhabdoviridae virus at a low M.O.I. in an original volume or after increasing cell density,
- (c) incubating the infected cells to produce an infected cell culture supernatant, and
- (d) harvesting said infected cell culture supernatant.
A method for making a composition comprising Rhabdoviridae virus, wherein said composition has a volume greater than 300 mL and a Rhabdoviridae virus titer greater than 108 TCID50 per mL, said method comprising:
- (a) obtaining a sample of Rhabdoviridae virus in serum-free medium, and
- (b) obtaining Rhabdoviridae virus from said sample to form said composition without performing an anion exchange step.
A method for assessing a composition comprising Rhabdoviridae viruses for the presence of non-Rhabdoviridae viruses, wherein said method comprises:
- (a) contacting said composition with an antibody preparation under conditions wherein said preparation neutralizes said Rhabdoviridae viruses within said composition thereby forming a neutralized Rhabdoviridae virus sample,
- (b) incubating said sample with viable cells, and
- (c) determining whether or not said cells exhibit a cytopathic effect, wherein the presence of said cytopathic effect indicates that said composition comprises non-Rhabdoviridae viruses, and wherein the absence of said cytopathic effect indicates that said composition lacks non-Rhabdoviridae viruses.
Dependencies are inferred from the claim text.