PCV/mycoplasma hyopneumoniae/PRRS combination vaccine

1. A method for preparing a multivalent immunogenic composition including a soluble portion of a Mycoplasma hyopneumoniae (M.hyo) whole cell preparation, the method comprising:

i) culturing M.hyo in a suitable media over periods ranging from 18-144 hours;

ii) subsequently inactivating the M.hyo culture;

iii) harvesting the inactivated culture fluid, wherein the inactivated culture fluid comprises an M.hyo whole cell preparation comprising both a soluble liquid fraction and insoluble cellular material;

iv) separating the soluble liquid fraction from the insoluble cellular material;

v) substantially removing both IgG and antigen/immunoglobulin immunocomplexes from the separated soluble liquid fraction to form a soluble portion of the M.hyo whole cell preparation; and

vi) subsequently combining the soluble portion of the M.hyo whole cell preparation with a PCV2 antigen and PRRS virus antigen.

2. The method of claim 1 , wherein vi) comprises combining a ready-to-use liquid composition comprising both the PCV2 antigen and the M.hyo soluble portion with a lyophilized PRRS virus antigen.

3. The method of claim 1 , wherein the M.hyo culture is inactivated by exposing the culture to an inactivation agent.

4. The method of claim 3 , wherein the inactivation agent is binary ethylenimine (BEI).

5. The method of claim 3 , further comprising neutralizing the inactivation agent.

6. The method of claim 1 , wherein the soluble liquid fraction is separated from the insoluble cellular material by a centrifugation step.

7. The method of claim 1 , wherein the soluble liquid fraction is separated from the insoluble cellular material by a filtration step.

8. The method of claim 1 , wherein the soluble liquid fraction is separated from the insoluble cellular material by a precipitation step.

9. The method of claim 1 , wherein the separated soluble liquid fraction is treated with a Protein A or Protein G resin to substantially remove both the IgG and antigen/immunoglobulin immunocomplexes contained in the separated soluble liquid fraction.

10. The method of claim 9 , wherein the separated soluble liquid fraction is treated with a Protein A resin to substantially remove both the IgG and antigen/immunoglobulin immunocomplexes contained in the separated soluble liquid fraction.

11. The method of claim 1 , further comprising admixing the immunogenic composition with an adjuvant.

12. The method of claim 11 , wherein the adjuvant is an oil emulsion comprising a polyoxyethylene-polyoxypropylene block copolymer, squalane, polyoxyethylene sorbitan monooleate and a buffered salt solution (SP-oil).

13. The method of claim 1 , further comprising admixing the immunogenic composition with a pharmaceutically-acceptable carrier.

14. The method of claim 1 , further comprising combining the immunogenic composition with at least one additional porcine antigen.