REVERSE GENETICS METHODS FOR VIRUS RESCUE
A method for rescuing a virus by reverse genetics is provided in which cells are added after transfection.
1 - 28 . (canceled)
29 . A method for preparing a viral composition, the method comprising the steps of:
(i) transfecting a culture eukaryotic cells with at least one expression construct encoding a viral RNA molecule;
(ii) adding untransfected eukaryotic cells to the transfected cells of step (i) to provide a mixture of cells, wherein the untransfected cells are of the same type as the cells of step (i);
(iii) culturing the mixture of cells from step (ii) so as to produce viruses in the culture; and,
(iv) harvesting viruses, proteins or combination thereof from the culture obtained in step (iii), so as to obtain a viral composition.
30 . The method of claim 29 , further comprising a step of:
(v) purifying the viruses from step (iv) so as to obtain purified virions.
31 . The method of claim 30 , wherein the virions obtained in step (v) is whole virions.
32 . The method of claim 30 , further comprising inactivating the viruses or the purified virions.
33 . The method of claim 31 , further comprising packaging the whole virions in a sterile container.
34 . The method of claim 33 , wherein the viral composition further comprises an adjuvant.
35 . A kit comprising the whole virions of claim 33 .
36 . The kit of claim 34 , further comprising an adjuvant component provided in a separate container.
37 . The method of claim 30 , further comprising a step of:
treating the purified virions with a detergent so as to obtain split virions.
38 . The method of claim 37 , further comprising inactivating the split virions.
39 . The method of claim 37 , further comprising packaging the split virions in a sterile container.
40 . The method of claim 39 , wherein the viral composition further comprises an adjuvant.
41 . A kit comprising the split virions of claim 37 .
42 . The kit of claim 40 , further comprising an adjuvant component provided in a separate container.
43 . The method of claim 29 , wherein the proteins of step (iv) are one or more surface antigens.
44 . The method of claim 43 , further comprising a step of:
purifying the one or more surface antigens so as to obtain purified surface antigens.
45 . The method of claim 43 , further comprising inactivating the surface antigens.
46 . The method of claim 44 , further comprising packaging the purified surface antigens in a sterile container.
47 . The method of claim 44 , wherein the viral composition further comprises an adjuvant.
48 . A kit comprising the surface antigens of claim 46 .
49 . The kit of claim 47 , further comprising an adjuvant component provided in a separate container.