CLIAMS:1. A scalable method for purification of immunogenic tetanus toxoid, said method comprising the steps of:
a. obtaining an immunogenic tetanus toxoid containing material prepared from fermentation of Clostridium tetani;
b. carrying out first step of separation of tetanus toxoid comprising the steps of:
i. incubating said tetanus toxoid containing material with ammonium sulphate for 12-16 hours at 2-8ºC under mild stirring;
ii. obtain tetanus toxoid salt precipitate by centrifuging; and
iii. dissolving said tetanus toxoid salt precipitate in a solvent to obtain a diluted clear solution.
c. carrying out a second step of separation using said diluted solution using a tangential flow filtration setup having membrane pore size in the range of 10-30kDa to obtain a retentate comprising of purified immunogenic tetanus toxoid,
wherein purity of tetanus toxoid is in the range of 1600-1900Lf/mg pN, and wherein percentage recovery of said purified tetanus toxoid is in the range of 70-95%.
2. The method as claimed in claim 1, wherein ammonium sulphate w/v% is in the range of 15-30%.
3. The method as claimed in any of the claims 1-2, wherein said purified immunogenic tetanus toxoid is optionally further filter sterilized with a 0.2µm filter.
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