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US 10,208,291 B2Granted
Method for generating high-titer hepatitis E virus stocks and titration assay for hepatitis E virus
AssigneeGrifols, S.A.
InventorsBrett Buno, Terri Journigan, Joann Hotta, Michael Burdick
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Abstract
Polybrene as an additive in cell culture medium is used in methods for the generation of high-titer hepatitis E virus stocks and assays for titration of hepatitis E virus. A cell culture medium containing polybrene is used for high-titer HEV generation, a method for determining the presence and/or the level of HEV in a sample, and an HEV titration assay using polybrene.
Claims13
A method of producing a titer of Hepatitis E Virus (HEV), the method comprising:
- a) culturing a cell line in vitro in a medium comprising a concentration of polybrene, and
- b) infecting the cell line with HEV,
- wherein the cell line is HepG2 (ATCC number HB-8065) or HepG2/C3A (ATCC number CRL-10741),
- wherein the concentration of polybrene is about 1 μg/ml - about 5 μg/ml.
1 more element
A method of determining a presence and/or a level of HEV in a sample, the method comprising the steps of:
- a) providing the sample to a mixture comprising a cell line and a culture medium, wherein the culture medium comprises polybrene;
- b) incubating the mixture comprising the sample from step a), to allow for the propagation of HEV, if present in the sample;
- c) collecting a portion of step b), the portion comprising HEV, if present and propagated during step b);
- and
- d) measuring the presence and/or the level of a biological substance associated with HEV in the collected portion.
2 more elements
Dependencies are inferred from the claim text.