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COMPARISON OF GEL FILTRATION AND AMMONIUM SULPHATE PRECIPITATION IN THE PURIFICATION OF DIPHTHERIA TOXIN AND TOXOID
Author(s) -
MØYNER KIRSTEN,
CHRISTIANSEN GERD
Publication year - 1984
Publication title -
acta pathologica microbiologica scandinavica series c: immunology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.909
H-Index - 88
eISSN - 1600-0463
pISSN - 0108-0202
DOI - 10.1111/j.1699-0463.1984.tb00047.x
Subject(s) - diphtheria toxin , chromatography , toxoid , size exclusion chromatography , chemistry , precipitation , ammonium , ammonium sulfate , filtration (mathematics) , flocculation , fraction (chemistry) , ammonium sulfate precipitation , toxin , corynebacterium diphtheriae , diphtheria , biochemistry , biology , immunization , antibody , immunology , organic chemistry , physics , statistics , mathematics , vaccination , meteorology , enzyme
Crude diphtheria toxin and toxoid were subjected to purification by gel filtration and stepwise ammonium sulphate precipitation. The various fractions obtained by the purification procedures were studied by immunological methods. A high molecular weight fraction of glycoprotein nature was present in all of the crude preparations studied. The fraction was antigenically non‐identical with the real toxin or toxoid and did not have its origin in the culture medium. It showed a long flocculation time when tested against equine diphtheria toxoid antiserum. The fraction could be removed from the crude preparations by gel filtration or by precipitation with 21% (w/v) ammonium sulphate. When comparing toxoids purified by each of these methods, the method of gel filtration resulted in a somewhat higher degree of purity, suggesting that this method would be more suitable than the AS precipitation method for the purification of diphtheria toxoid.