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 method for the determination of an antigen content of a porcine circovirus type 2 (PCV-2) antigen in a mixture of at least a first composition comprising the PCV-2 antigen and a second composition comprising (i) a Mycoplasma hyopneumoniae ( M. hyo ) antigen and (ii) antibodies directed against the PCV-2 antigen that are capable of binding with the PCV-2 antigen, the method comprising the steps of,
a) mixing together the first and second compositions,
b) dissociating antigen-antibody complexes in a sample of the mixture, formed between the PCV-2 antigen and the antibodies, and
c) determining the antigen content of the PCV-2 antigen by means of an immunoassay.
2. The method of claim 1 , wherein the immunoassay is an ELISA (enzyme linked immunosorbent assay).
3. The method of claim 1 , wherein the mixture is a ready-to-use vaccine formulation.
4. The method of claim 1 , wherein the sample of the mixture is incubated with an acid solution to dissociate the antigen-antibody complexes.
5. The method of claim 4 , wherein the acid solution is a citric acid solution.
6. The method of claim 4 , wherein the sample of the mixture is incubated with the acid solution for at least 8 hours.
7. The method of claim 4 , wherein the sample of the mixture is incubated at a ratio (v/v) between the acid solution and the mixture selected from the group consisting of at least 25, 25-75, and 25-50.
8. The method of claim 4 , wherein the pH range of the acid solution is pH 1.0-3.0.
9. The method of claim 1 , wherein the method is a potency test of a combination vaccine.
10. The method of claim 3 , wherein the sample of the mixture is incubated with an acid solution to dissociate the antigen-antibody complexes.
11. The method of claim 10 , wherein the acid solution is a citric acid solution.
12. The method of claim 10 , wherein the sample of the mixture is incubated with the acid solution for at least 8 hours.
13. The method of claim 10 , wherein the sample of the mixture is incubated at a ratio (v/v) between the acid solution and the mixture selected from the group consisting of at least 25, 25-75, and 25-50.
14. The method of claim 10 , wherein the pH range of the acid solution is pH 1.0-3.0.
15. The method of claim 2 , wherein the sample of the mixture is incubated with an acid solution to dissociate the antigen-antibody complexes.
16. The method of claim 15 , wherein the acid solution is a citric acid solution.
17. The method of claim 15 , wherein the sample of the mixture is incubated with the acid solution for at least 8 hours.
18. The method of claim 15 , wherein the sample of the mixture is incubated at a ratio (v/v) between the acid solution and the mixture selected from the group consisting of at least 25, 25-75, and 25-50.
19. The method of claim 15 , wherein the pH range of the acid solution is pH 1.0-3.0.