Method of preparing a PRRS virus vaccine
View Patent ↗This invention provides kits, devices, and methods for the detection of antibodies that recognize one or more proteins and/or antigens from porcine reproductive and respiratory syndrome virus (PRRSV). The antibodies may be in a biological fluid of a PRRSV infected or at risk subject. The invention may be advantageously applied to both the diagnosis and prevention of PRRSV infection.
1. A method of preparing a vaccine composition containing PRRS virus proteins and antigens from PRRS virus infected cells, said method comprising
a) providing a population of cells that have been infected with PRRS virus, wherein said cells produce viral proteins and antigens encoded by PRRS virus so that the majority of viral proteins and antigens encoded by the PRRS virus are either within the infected cells or associated with the cell membrane of the infected cells;
b) isolating the PRRS virus infected cells, away from cell-free PRRS virus in the culture medium about the cells, to form isolated cells containing cell associated viral proteins and antigens encoded by PRRS virus;
c) extracting or eluting the isolated cells with a detergent containing solution to form an extract or eluate containing the cell associated viral proteins and antigens along with cellular components; and
d) combining said extract or eluate with an adjuvant to form a vaccine composition that protects a pig against PRRS virus infection.
2. The method of claim 1 wherein said detergent containing solution contains a non-ionic detergent.
3. The method of claim 1 wherein said extracting or eluting is for about 2 to about 15 hours at 4° C.
4. The method of claim 1 wherein said proteins and antigens include PRRS virus envelope proteins or the N protein.
5. The method of claim 1 wherein said proteins and antigens include a PRRS virus encoded glycoprotein.
6. The method of claim 1 wherein said proteins and antigens include a PRRS virus non-structural protein.
7. A method of preparing a vaccine composition containing PRRS virus proteins and antigens from PRRS virus infected cells, said method comprising
a) providing a population of cells that have been infected with PRRS virus, wherein said cells produce viral proteins and antigens encoded by PRRS virus so that said cells produce cell-free PRRS virus into the culture medium;
b) isolating the PRRS virus infected cells, away from cell-free PRRS virus in the culture medium about the cells, to form isolated cells containing cell associated viral proteins and antigens encoded by PRRS virus;
c) extracting or eluting the isolated cells with a detergent containing solution to form an extract or eluate containing the cell associated viral proteins and antigens along with cellular components; and
d) combining said extract or eluate with an adjuvant to form a vaccine composition that protects a pig against PRRS virus infection.
8. The method of claim 7 wherein said detergent containing solution contains a non-ionic detergent.
9. The method of claim 7 wherein said extracting or eluting is for about 2 to about 15 hours at 4° C.
10. The method of claim 7 wherein said proteins and antigens include PRRS virus envelope proteins or the N protein.
11. The method of claim 7 wherein said proteins and antigens include a PRRS virus encoded glycoprotein.
12. The method of claim 7 wherein said proteins and antigens include a PRRS virus non-structural protein.