1-17. (canceled)
18. A method, within an application server, of modifying an intra-transactional cache, comprising:
determining whether a copy of an original first data entity, accessed in the intra-transactional cache by a first client, has been updated;
determining whether the copy is also within an inter-transactional cache;
upon the copy not being within the inter-transactional cache, adding the copy to the inter-transactional cache; and
after the copy has been added to the inter-transactional cache, removing, from the intra-transactional cache, the copy.
19. The method of claim 18, further comprising:
upon a determination that the copy has been updated,
changing a version identifier associated with the original first data entity,
generating a raw data object based upon the updated copy, and
writing the raw data object to a repository separate from both the intra-transactional cache and the inter-transactional cache.
20. The method of claim 18, further comprising:
removing, from a data entry list for the copy within the intra-transactional cache, a client ID and lock entry.
21. The method of claim 18, wherein
both the intra-transactional cache and the inter-transactional cache are accessible by the application server.
22. The method of claim 18, wherein
both the intra-transactional cache and the inter-transactional cache are within the application server.
23. A method, within an application server, of modifying an intra-transactional cache, comprising:
determining whether a copy of an original first data entity, accessed in the intra-transactional cache by a first client, has been updated;
determining whether the copy is also within an inter-transactional cache;
upon a determination that the copy is within the inter-transactional cache, determining whether a lock exists for the copy within the inter-transactional cache; and
removing, from the intra-transactional cache, the copy.
24. The method of claim 23, further comprising:
upon a determination that the copy has been updated,
changing a version identifier associated with the original first data entity,
generating a raw data object based upon the updated copy, and
writing the raw data object to a repository separate from both the intra-transactional cache and the inter-transactional cache.
25. The method of claim 23, further comprising:
upon a determination that no lock exists for the copy within the inter-transactional cache, updating the copy within the inter-transactional cache.
26. The method of claim 23, wherein
both the intra-transactional cache and the inter-transactional cache are accessible by the application server.
27. The method of claim 23, further comprising:
removing, from a data entry list for the copy within the intra-transactional cache, a client ID and lock entry.
28. A computer program product comprising computer usable storage medium having stored therein computer usable program code for modifying an intra-transactional cache, the computer usable program code, which when executed by an application server, causes the application server to perform:
determining whether a copy of an original first data entity, accessed in the intra-transactional cache by a first client, has been updated;
determining whether the copy is also within an inter-transactional cache;
upon the copy not being within the inter-transactional cache, adding the copy to the inter-transactional cache; and
after the copy has been added to the inter-transactional cache, removing, from the intra-transactional cache, the copy.
29. The computer program product of claim 28, wherein the computer usable program code further causes the application server to perform:
upon a determination that the copy has been updated,
changing a version identifier associated with the original first data entity,
generating a raw data object based upon the updated copy, and
writing the raw data object to a repository separate from both the intra-transactional cache and the inter-transactional cache.
30. The computer program product of claim 28, wherein the computer usable program code further causes the application server to perform:
removing, from a data entry list for the copy within the intra-transactional cache, a client ID and lock entry.
31. The computer program product of claim 28, wherein
both the intra-transactional cache and the inter-transactional cache are accessible by the application server.
32. The computer program product of claim 28, wherein
both the intra-transactional cache and the inter-transactional cache are within the application server.
33. A computer program product comprising computer usable storage medium having stored therein computer usable program code for modifying an intra-transactional cache, the computer usable program code, which when executed by an application server, causes the application server to perform:
determining whether a copy of an original first data entity, accessed in the intra-transactional cache by a first client, has been updated;
determining whether the copy is also within an inter-transactional cache;
upon a determination that the copy is within the inter-transactional cache, determining whether a lock exists for the copy within the inter-transactional cache; and
removing, from the intra-transactional cache, the copy.
34. The computer program product of claim 33, wherein the computer usable program code further causes the application server to perform:
upon a determination that the copy has been updated,
changing a version identifier associated with the original first data entity,
generating a raw data object based upon the updated copy, and
writing the raw data object to a repository separate from both the intra-transactional cache and the inter-transactional cache.
35. The computer program product of claim 33, wherein the computer usable program code further causes the application server to perform:
upon a determination that no lock exists for the copy within the inter-transactional cache, updating the copy within the inter-transactional cache.
36. The computer program product of claim 33, wherein
both the intra-transactional cache and the inter-transactional cache are accessible by the application server.
37. The computer program product of claim 33, wherein the computer usable program code further causes the application server to perform:
removing, from a data entry list for the copy within the intra-transactional cache, a client ID and lock entry.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
We claim:
1. A method of producing virus from a culture of cells, comprising the steps of:
(a) providing a culture of cells which has been infected by the virus;
(b) extracting the virus from the cells by adding a detergent to the culture and incubating for a period of time to result in a cell lysate;
(c) removing cell debris; and
(d) collecting the virus.
2. The method of claim 1 wherein the cell debris is removed by filtration.
3. The method of claim 1 wherein the cell debris is removed by step-wise filtration comprising:
(1) filtering through a prefilter having a pore size of 5 M or 8 M, and
(2) filtering after step (1) through a combination filter having pore sizes of 3 M and 0.8 M.
4. The method of claim 1 further comprising treating the cell lysate with benzonase.
5. The method of claim 2 further comprising concentrating the filtrate.
6. The method of claim 5 wherein the filtrate is concentrated by diafiltration.
7. The method of claim 1 wherein the virus is a non-enveloped virus.
8. The method of claim 1 wherein the virus is a reovirus.
9. The method of claim 8 wherein the reovirus is a mammalian reovirus.
10. The method of claim 9 wherein the mammalian reovirus is a human reovirus.
11. The method of claim 10 wherein the human reovirus is a serotype 3 virus.
12. The method of claim 11 wherein the serotype 3 reovirus is the Dearing strain.
13. The method of claim 1 wherein the reovirus is a recombinant reovirus.
14. The method of claim 1 wherein the cells are human embryo kidney 293 (HEK 293) cells.
15. The method of claim 14 wherein the HEK 293 cells are grown in suspension.
16. The method of claim 1 further comprising purifying the virus by anion exchange chromatography.
17. The method of claim 1 further comprising purifying the virus by a combination of ion exchange and size exclusion chromatography
18. A composition comprising the virus collected according to claim 1.
19. The composition of claim 18 which is suitable for clinical administration.
20. The composition of claim 19 further comprising a pharmaceutically acceptable excipient andor carrier.
21. A method of producing infectious reovirus, comprising:
(a) providing a culture of HEK 293 cells which has been infected by reovirus;
(b) extracting the virus from the cells by adding Triton X-100 to the culture and incubating at about 25 C. to about 37 C.;
(c) treating the mixture from step (b) with benzonase;
(d) removing cell debris by filtration;
(e) concentrating the filtrate by ultrafiltration or diafiltration;
(f) purifying the reovirus by a combination of ion exchange and size exclusion chromatography; and
(g) collecting the reovirus.
22. A composition comprising the reovirus prepared according to the method of claim 21.
23. The composition of claim 22 further comprising a pharmaceutically acceptable excipient andor carrier.