In-vitro potency assay for protein-based meningococcal vaccines

Abstract

The invention uses ELISA or similar assays for analysing a meningococcal vaccine. The assay uses antibodies which bind to meningococcal proteins within the vaccine, and in particular monoclonal antibodies which are bactericidal for meningococcus and/or which recognise conformational epitopes within the meningococcal proteins. By performing the assay on a series of dilutions of a test vaccine, and by comparing the results with those obtained using a reference vaccine of known potency, it is possible to determine the relative potency of the test vaccine. This value can be used as a parameter for determining whether a manufactured batch of a vaccine is suitable for release to the public, or whether it has experienced a production failure and so should not be used.

1 - 18 . (canceled) 
     
     
         19 . A kit comprising:
 (i) an anti-vaccine monoclonal antibody;   (ii) an immobilized antigen which is recognized by the anti-vaccine monoclonal antibody, wherein the anti-vaccine monoclonal antibody (a) is bactericidal for meningococcus or (b) recognizes a conformational epitope in the meningococcal antigen.   
     
     
         20 . The kit of  claim 19 , further comprising a labeled antibody which binds to the anti-vaccine monoclonal antibody. 
     
     
         21 . The kit of  claim 19 , wherein the immobilized antigen is from a meningococcal protein, a fusion protein comprising a meningococcal protein, or a truncated form of a meningococcal protein. 
     
     
         22 . The kit of  claim 21 , wherein the immobilized antigen is meningococcal Neisserial Heparin Binding Antigen (NHBA), meningococcal factor H binding protein (fHbp), or meningococcal Neisserial adhesin A (NadA). 
     
     
         23 . The kit of  claim 20 , wherein the labeled antibody is labelled with an enzyme. 
     
     
         24 . The kit of  claim 23 , wherein the enzyme is a peroxidase, a phosphatase, a laccase or a beta-galactosidase. 
     
     
         25 . The kit of  claim 19 , wherein the anti-vaccine monoclonal antibody comprises a monoclonal antibody with a variable light (VL) region comprising the amino acid sequence of SEQ ID NO:21. 
     
     
         26 . The kit of  claim 19 , wherein the anti-vaccine monoclonal antibody comprises a monoclonal antibody with a variable heavy (VH) region comprising the amino acid sequence of SEQ ID NO:22. 
     
     
         27 . The kit of  claim 19 , wherein the anti-vaccine monoclonal antibody comprises a monoclonal antibody with variable light (VL) and variable heavy (VH) regions comprising the amino acid sequences of SEQ ID NO:21 and SEQ ID NO:22. 
     
     
         28 . The kit of  claim 19 , wherein the anti-vaccine monoclonal antibody comprises CDRs from the variable light (VL) and variable heavy (VH) regions of SEQ ID NO:21 and SEQ ID NO:22. 
     
     
         29 . The kit of  claim 19 , wherein the anti-vaccine monoclonal antibody is a murine monoclonal IgG antibody. 
     
     
         30 . The kit of  claim 29 , wherein the murine monoclonal IgG antibody is a murine monoclonal IgG2b antibody. 
     
     
         31 . The kit of  claim 30 , wherein the immobilized meningococcal antigen is fHbp. 
     
     
         32 . The kit of  claim 19 , wherein the anti-vaccine monoclonal antibody comprises a high-affinity tag. 
     
     
         33 . The kit of  claim 32 , wherein the high-affinity tag is biotin, avidin or streptavidin. 
     
     
         34 . The kit of  claim 33 , further comprising an enzyme conjugated to a ligand of the high affinity tag. 
     
     
         35 . The kit of  claim 19 , wherein the immobilized meningococcal antigen is immobilized on a surface. 
     
     
         36 . The kit of  claim 19 , further comprising a buffer and a microwell plate, wherein the immobilized meningococcal antigen is immobilized on the microwell plate.