METHOD FOR THE PREPARATION OF DIPHTHERIA TOXOID VACCINE

摘要

A method for the preparation of diphtheria toxoid vaccine is provided to improve preparation yield of diphtheria toxin by optimizing the cultivation condition, and reduce purification costs and improve purity of diphtheria toxin by using ammonium sulfate treatment and ion exchange chromatography. The method for the preparation of diphtheria toxoid vaccine comprises the steps of: culturing Corynebacterium diphtheriae in medium for 24-72 hours by supplying 0.1-1 vvm of a mixture containing air and oxygen into the fermenter to maintain dissolved oxygen amount of the medium at 10-50%, adding 1-30g/l of maltose into the medium after 10-30 hours of the cultivation, and further culturing it; centrifuging the cultured medium, separating the supernatant and concentrating the supernatant with ultrafiltration and diafiltration; precipitating the concentrated diphtheria toxin with ammonium sulfate and dialyzing the precipitates with 5-8 mS/cm of phosphate buffer; purifying the crude purified diphtheria toxin with column chromatography using 5-8 mS/cm of phosphate buffer; and removing toxicity of diphtheria toxin with formalin.