Modified human rotaviruses and uses therefor
AssigneeMurdoch Childrens Research Institute
InventorsCarl Kirkwood, Ruth Frances Bishop, Graeme Laurence Barnes
Life of this patent
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Abstract
The present disclosure relates generally to the field of viral vaccines. Particularly, the present disclosure provides a modified Vero-adapted human rotavirus strain and a culturing method to produce high titer virus, a rotavirus vaccine, vaccination protocols and diagnostics and prognostic assays.
Claims20
A method for culturing a strain of rotavirus (RV), said method comprising subjecting an aliquot or inoculum of RV to activation by a trypsinization step comprising administering trypsin at greater than 3 times the standard concentration of trypsin used for activation of RV for infection into cells of 5-15 μg/ml and culturing the activated RV in Vero cells or a Vero cell-derived cell line for a time and under conditions sufficient for the RV to undergo a genetic modification resulting in an amino acid change in one or more of VP1, VP3, VP4, NSP4 and/or NSP5/6 and/or nucleotide change in one or more of VP1, VP3, VP4, NSP2, NSP4 and/or NSP5/6.
A method for generating an attenuated RV3 in Vero cells, said method comprising subjecting an aliquot or inoculum of RV3 to activation by a trypsinization step comprising administering trypsin at greater than 3 times the standard concentration of trypsin used for activation of RV3 for infection into cells of 5-15 μg/ml and culturing the activated RV3 and culturing the activated RV3 with Vero cells or a Vero cell-derived cell line for a time and under conditions sufficient for a genetic modification to occur selected from the list provided in Table 2 or Table 5 compared to MA104-adapted RV3.
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Dependencies are inferred from the claim text.