Purification of HBV antigens for use in vaccines

Abstract

The present invention relates to a method for the production of a hepatitis B antigen suitable for use in a vaccine, the method comprising purification of the antigen in the presence of cysteine, to vaccines comprising such antigens.

1 . A method for making a hepatitis B antigen for use in a vaccine, the method comprising purification of the hepatitis B antigen in the presence of a reducing agent having a free —SH group.  
   
   
       2 . The method according to  claim 1  wherein the hepatitis B antigen is a stable immunogenic antigen free of thiomersal.  
   
   
       3 . The method according to  claim 1  wherein the purified antigen product comprises less than about 0.025 μg mercury per 20 kg protein.  
   
   
       4 . The method according to  claim 1  wherein the purified antigen product comprises no thiomersal.  
   
   
       5 . The method of producing a hepatitis B antigen according to  claim 1  wherein a crude hepatitis B antigen preparation is: 
 (a) subjected to gel permeation chromatography;    (b) subjected to ion-exchange chromatography; and    (c) mixed with a reducing agent having a free —SH group.    
   
   
       6 . The method according to  claim 1  wherein the reducing agent is chosen from the group of: cysteine, glutathione, dithiothreitol and β-mercaptoethanol.  
   
   
       7 . The method according to  claim 1 , wherein the reducing agent is cysteine.  
   
   
       8 . The method according to  claim 7  wherein the cysteine is added to a final concentration of between about 1-10 mM.  
   
   
       9 . The method according to  claim 7  wherein the cysteine is added to a final concentration of about 2 mM.  
   
   
       10 . A vaccine composition comprising a hepatitis B antigen in the presence of a reducing agent having a free —SH group.  
   
   
       11 . The vaccine composition according to  claim 10  further comprising an adjuvant.  
   
   
       12 . The vaccine composition according to  claim 11  wherein the adjuvant is an aluminium salt.  
   
   
       13 . The vaccine composition as claimed in  claim 12 , which further comprises a TH-1 inducing adjuvant.  
   
   
       14 . The vaccine composition according to  claim 13  wherein the TH1-inducing adjuvant is chosen from the group of: 3-DMPL, QS21, 3-DMPL and QS21, and a CpG oligonucleotide.  
   
   
       15 . The vaccine composition as claimed in  claim 10 , which further comprises at least one of the antigens chosen from the group of: diptheria toxoid (D), tetanus toxoid (T) acellular pertussis antigens (Pa), inactivated polio virus (IPV),  haemophilus influenzae  antigen (Hib), hepatitis A antigen, herpes simplex virus (HSV), chlamydia, GSB, HPV,  streptococcus pneumoniae , and neisseria antigens.  
   
   
       16 . The vaccine composition comprising a hepatitis B antigen and a cysteine solution.  
   
   
       17 . A vaccine composition comprising a hepatitis B antigen, an adjuvant, and an inactivated polio virus.