Potency test for vaccine formulations
The invention relates to certain methods for the determination of an antigen content of a first antigen in a mixture comprising two or more antigens. The invention also relates to a potency test for an antigen in a combination vaccine. The method allows the determination of the antigen content in a mixture additionally comprising antibodies that are capable of binding with the antigen.
1. A composition comprising a porcine circovirus type 2 (PCV-2) antigen and a Mycoplasma hyopneumoniae ( M. hyo ) antigen;
wherein the M. hyo antigen originates from media comprising serum that also contains an antibody that is capable of binding to the PCV-2 antigen; wherein the M. hyo antigen has been separated from said antibody; and wherein said composition is obtained by the method of:
A. separating the M. hyo antigen from said antibody by either centrifugation or an immuno-depletion step; wherein the M. hyo antigen is separated from said antibody; and
B. mixing the PCV-2 antigen with the M. hyo antigen separated from said antibody.
2. A vaccine comprising the composition of claim 1 ; wherein the PCV-2 antigen is a PCV-2 ORF2 protein, the M. hyo antigen is a bacterin, or both the PCV-2 antigen is a PCV-2 ORF2 protein and the M. hyo antigen is a bacterin.
3. A mixture comprising a porcine circovirus type 2 (PCV-2) antigen and a Mycoplasma hyopneumoniae ( M hyo ) antigen; wherein the M hyo antigen originates from media comprising serum that also contains an antibody that is capable of binding to the PCV-2 antigen; and wherein the M hyo antigen had been separated from said antibody prior to mixing the M hyo antigen with the PCV-2 antigen to form said mixture.
4. The vaccine of claim 2 wherein the M. hyo antigen is an M hyo bacterin and the PCV-2 antigen is a PCV-2 ORF2 protein.
5. The composition of claim 1 , wherein said separating of the M. hyo antigen from said antibody is performed by an immuno-depletion step.
6. The composition of claim 5 , wherein said immuno-depletion step is performed with a Protein-G ligand or a Protein-A ligand.
7. The composition of claim 6 , wherein the Protein-G ligand or the Protein-A ligand is bound to a solid phase.
8. The composition of claim 5 , wherein said immuno-depletion step is performed with the Protein-A ligand.
9. The composition of claim 8 , wherein the Protein-A ligand is bound to a solid phase.
10. A method for determining the antigen content of a porcine circovirus type 2 (PCV-2) antigen in a mixture comprising the PCV-2 antigen and a Mycoplasma hyopneumoniae ( M hyo ) antigen; wherein the M. hyo antigen originates from media comprising serum that also contains an antibody that is capable of binding to the PCV-2 antigen; wherein the method comprises the steps of:
A. separating the M hyo antigen from said antibody;
B. mixing the M. hyo antigen with the PCV-2 antigen to form a mixture; and
C. determining the antigen content of the PCV-2 antigen in the mixture by means of an immunoassay.
11. The method of claim 10 , wherein the method is a potency test of a combination vaccine.
12. The method of claim 10 , wherein said separating is performed by centrifugation or by an immune-depletion step.
13. The method of claim 12 , wherein said separating is performed by immune-depletion, comprising the steps of:
(i) capturing said antibody with a ligand that exhibits an affinity for said antibody, whereby a ligand-antibody complex is formed; and
(ii) separating the ligand-antibody complex from the M. hyo antigen.
14. The method of claim 13 , wherein the ligand is bound to a solid phase.
15. The method of claim 10 , wherein the PCV-2 antigen is a PCV-2 ORF2 protein.
16. A method for preparing a composition comprising a porcine circovirus type 2 (PCV-2) antigen and a Mycoplasma hyopneumoniae ( M hyo ) antigen, wherein the M. hyo antigen originates from media comprising serum that also contains an antibody that is capable of binding with the PCV-2 antigen, said method comprising the steps of:
A. separating the M hyo antigen from said antibody; and
B. mixing the M. hyo antigen with the PCV-2 antigen.
17. The method of claim 16 , wherein said separating is performed by centrifugation or by an immune-depletion step.
18. The method of claim 17 , wherein said separating is performed by immune-depletion, comprising the steps of:
(i) capturing said antibody with a ligand that exhibits an affinity for said antibody, whereby a ligand-antibody complex is formed; and
(ii) separating the ligand-antibody complex from the M. hyo antigen.
19. The method of claim 18 , wherein the ligand is bound to a solid phase.
20. The method of claim 19 wherein the PCV-2 antigen is a PCV-2 ORF2 protein.
21. The method of claim 15 , wherein the method is a potency test of a combination vaccine.
22. The method of claim 15 , wherein the M. hyo antigen is an M hyo bacterin.
23. The method of claim 13 , wherein said immuno-depletion step is performed with a Protein-G ligand or a Protein-A ligand.
24. The method of claim 23 , wherein said immuno-depletion step is performed with a Protein-A ligand.
25. The method of claim 24 , wherein the Protein-A ligand is bound to a solid phase.
26. The method of claim 20 , wherein the M hyo antigen is an M hyo bacterin.
27. The method of claim 18 , wherein the ligand is Protein-G or Protein-A.
28. The method of claim 27 , wherein the ligand is Protein-A.
29. A vaccine comprising the composition of claim 1 , wherein the PCV- 2 antigen is an inactivated PCV- 2 .