Potency test for vaccine formulations

Abstract

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 first antigen in a mixture of at least a composition comprising the first antigen and a composition comprising (i) a second antigen and (ii) antibodies that are capable of binding with the first antigen, the method comprising the steps of:
 a) dissociating antigen-antibody complexes in the mixture, formed between the first antigen and the antibodies; and   b) determining the antigen content of the first antigen by means of an immunoassay.   
     
     
         2 . The method of  claim 1 , wherein the first antigen is a porcine circovirus type 2 (PCV-2) antigen and the second antigen is a  Mycoplasma hyopneumoniae  antigen. 
     
     
         3 . The method of  claim 1 , wherein the immunoassay is an ELISA (enzyme linked immunosorbant assay). 
     
     
         4 . The method of  claim 1 , wherein the mixture is a ready-to-use vaccine formulation. 
     
     
         5 . The method of  claim 1 , wherein the mixture is incubated with an acid solution to dissociate the antigen-antibody complexes. 
     
     
         6 . The method of  claim 5 , wherein the acid solution is a citric acid solution. 
     
     
         7 . The method of  claim 5 , wherein the mixture is incubated with the acid solution for at least 8 hours. 
     
     
         8 . The method of  claim 5 , wherein the mixture is incubated at a ratio (v/v) between the acid solution and the mixture of at least 25, preferably 25-75, more preferably 25-50. 
     
     
         9 . The method of  claim 5 , wherein the acid solution has a pH of 1.0-3.0. 
     
     
         10 . 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 composition comprising the PCV-2 antigen and a composition comprising a  Mycoplasma hyopneumoniae  ( M. hyo ) antigen, the method comprising the steps of:
 a) mixing the two compositions; and   b) determining the antigen content of the PCV-2 antigen by means of an immunoassay, characterized in that the  M. hyo  antigen is obtained from a culture that does not comprise swine serum.   
     
     
         11 . A method for the determination of an antigen content of a first antigen in a mixture of at least a composition comprising the first antigen and a composition comprising a second antigen, the method comprising the steps of:
 a) separating the second antigen from antibodies that are capable of binding with the first antigen in a composition comprising the second antigen and the antibodies,   b) mixing the second antigen with a composition comprising the first antigen; and   c) determining the antigen content of the first antigen in the mixture by means of an immunoassay.   
     
     
         12 . The method of  claim 11 , wherein the method is a potency test of a combination vaccine. 
     
     
         13 . The method of  claim 10 , wherein the method is a potency test of a combination vaccine. 
     
     
         14 . The method of  claim 1 , wherein the method is a potency test of a combination vaccine.