Method for porcine circovirus production and PCV2 vaccines
This invention relates to a method for rapid production of PCV2 such that an optimum yield of the virus can be obtained. This invention also relates to the vaccine useful against PCV2 which includes the PCV2 propagated using such a method.
1. A method of producing porcine circovirus type 2 (PCV2) comprising the steps of (1) preparing PK15 (porcine kidney) cells for infection; (2) infecting the PK15 cells with PCV2; (3) culturing and expanding the infected PK15 cells through two or more passages, and (4) isolating or harvesting PCV2.
2. The method of claim 1 , wherein the infection of PCV2 is carried out in the absence of glucosamine.
3. The method of claim 1 , wherein the PK15 cells are trypsinized at the end of each passage.
4. The method of claim 1 , wherein the PK15 infected cells are incubated at 36±2° C.
5. The method of claim 4 , wherein the incubation period for viral growth ranges from 2-8 days.
6. The method of claim 1 , wherein the passages range from 2-10 passages.
7. The method of claim 1 , wherein the passages beyond the initial cell passage/infection are carried out without the addition of fresh PK15 cells.
8. The method of claim 1 , wherein the virus split ratio (VSR) used for the passages ranges from 4-9.
9. The method of claim 1 , wherein the PCV2 is harvested from the culture by sonication and subsequent removing of the resultant cell debris by centrifugation.
10. An inactivated PCV2 vaccine prepared by the steps of (1) preparing PK15 (porcine kidney) cells for infection; (2) infecting the PK15 cells with PCV2; (3) culturing and expanding the infected PK15 cells through passages, and (4) isolating or harvesting PCV2; wherein the PCV2 was inactivated by a chemical inactivation method using the inactivating compound selected from the group consisting of enzymes, formaldehyde, beta-propiolactone (BPL), ethylene-imine, binary ethylenimine (BEI) and a derivative thereof, and wherein the PCV2 comprises a polynucleotide encoding a PCV2 ORF2 comprising SEQ ID NO:2, 4, or 6.
11. The inactivated PCV2 vaccine of claim 10 , wherein the infection of PCV2 is carried out in the absence of glucosamine.
12. The inactivated PCV2 vaccine of claim 10 , wherein the PK15 cells are trypsinized at the end of each passage.
13. The inactivated PCV2 vaccine of claim 10 , wherein the PK15 infected cells are incubated at 36±2° C.
14. The inactivated PCV2 vaccine of claim 10 , wherein the incubation period for viral growth ranges from 2-8 days.
15. The inactivated PCV2 vaccine of claim 10 , wherein the passages range from 2-10 passages.
16. The inactivated PCV2 vaccine of claim 10 , wherein the passages beyond the initial cell passage/infection are carried out without the addition of fresh PK15 cells.
17. The inactivated PCV2 vaccine of claim 10 , wherein the virus split ratio (VSR) used for the passages ranges from 4-9.
18. The inactivated PCV2 vaccine of claim 10 , wherein the PCV2 is harvested from the culture by sonication and subsequent removing of the resultant cell debris by centrifugation.
19. The inactivated PCV2 vaccine of claim 10 , wherein the vaccine further comprises one or more swine pathogens or viruses.
20. The inactivated PCV2 vaccine of claim 10 , wherein the polynucleotide comprises SEQ ID NO:1, 3, or 5.
21. The inactivated PCV2 vaccine of claim 10 , further comprises a pharmaceutically or veterinarily acceptable carriers, excipients, adjuvants, or stabilizers.