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US 7,598,072 B2Granted
Assay to detect viral uncoating
AssigneeWisconsin Alumni Research Foundation
InventorsJohn A. T. Young, Shakti Narayan
Life of this patent
Estimating the term…
Abstract
A cell-free viral uncoating assay is provided.
Claims16
A cell-free method for release of viral cores of enveloped viruses from endosomes, comprising:
- a) providing a cell-free sample comprising intact endosomes comprising fusion arrested enveloped viral particles, which endosomes are isolated from cells that express one or more viral receptors including the viral receptor TVA and are infected with a pseudotyped enveloped virus having a subgroup A ASLV envelope protein, wherein the endosomes are isolated in the presence of an amount of a lysosomotropic agent that elevates endosomal pH effective to arrest membrane fusion;
- and
- b) subjecting the sample to conditions that allow for virus particle-endosomal membrane fusion and release of viral cores from endosomes, yielding a composition.
A method to prepare a cell-free sample comprising isolated endosomes having fusion arrested enveloped viral particles, comprising:
- a) infecting cells that express one or more viral receptors including the viral receptor TVA with a pseudotyped enveloped virus having a subgroup A ASLV envelope protein in the presence of an amount of a lysosomotropic agent that elevates endosomal pH effective to arrest membrane fusion;
- and
- b) isolating endosomes from the infected cells.
Dependencies are inferred from the claim text.