Open AccessProcedures for large‐scale production and purification of Clostridium botulinum C1 toxin for preparation of toxoid
Author(s) -
Kurazono Hisao,
Shimozawa Kunio,
Hosokawa Masahiro,
Sakaguchi Genji
Publication year - 1985
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1985.tb00983.x
Subject(s) - sephadex , clostridium botulinum , chromatography , chemistry , protamine , protamine sulfate , ultrafiltration (renal) , toxin , clostridium tetani , size exclusion chromatography , biochemistry , microbiology and biotechnology , biology , enzyme , tetanus , virology , vaccination , heparin
Fortified cooked meat medium containing calcium carbonate (CaCO 3 ‐FCM) supported toxin production of a strain of Clostridium botulinum type C to a level of 2 × 10 6 mouse i.p. LD 50 /ml. C1 toxin was purified by sequential steps of acid precipitation from 5‐fold diluted culture supernatant in the presence of RNA, 2nd acid precipitation by dialysis, removal of RNA by protamine treatment, removal of excess protamine and bufferisation by ultrafiltration through Amicon PM‐30 membrane, sulphopropyl‐Sephadex chromatography, and Sephadex G‐200 gel filtration. By these procedures, 25 mg or more of highly purified C1 toxin was constantly obtained from a lot of 600‐ml culture.