Method for porcine circovirus production and PCV2 vaccines
View Patent ↗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. An inactivated PCV2 vaccine prepared by a method 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 passages, and (4) isolating or harvesting PCV2, wherein the PCV2 was inactivated by a physical inactivation method comprising subjecting the virus to energy-rich radiation selected from a group comprising UV light, X-radiation, and gamma-radiation, and wherein the PCV2 comprises a polynucleotide encoding a PCV2 ORF2 according to SEQ ID NO:2, 4, or 6.
2. The inactivated PCV2 vaccine of claim 1 , wherein the infection of PCV2 is carried out in the absence of glucosamine.
3. The inactivated PCV2 vaccine of claim 1 , wherein the PK15 cells are trypsinized at the end of each passage.
4. The inactivated PCV2 vaccine of claim 1 , wherein the PK15 infected cells are incubated at 36±2° C.
5. The inactivated PCV2 vaccine of claim 1 , wherein the incubation period for viral growth ranges from 2-8 days.
6. The inactivated PCV2 vaccine of claim 1 , wherein the passages range from 2-10 passages.
7. The inactivated PCV2 vaccine of claim 1 , wherein the passages beyond the initial cell passage/infection are carried out without the addition of fresh PK15 cells.
8. The inactivated PCV2 vaccine of claim 1 , wherein the virus split ratio (VSR) used for the passages ranges from 4-9.
9. The inactivated PCV2 vaccine of claim 1 , wherein the PCV2 is harvested from the culture by sonication and subsequent removing of the resultant cell debris by centrifugation.
10. The inactivated PCV2 vaccine of claim 1 , wherein the vaccine further comprises one or more swine pathogens or viruses.
11. The inactivated PCV2 vaccine of claim 1 , wherein the polynucleotide according to SEQ ID NO:1, 3, or 5.
12. The inactivated PCV2 vaccine of claim 1 , further comprises a pharmaceutically or veterinarily acceptable carriers, excipients, adjuvants, or stabilizers.