Abstract
Adverse reactions after injection of diphtheria vaccine are induced by impurities present in crude toxoids that cannot be removed completely by purification of toxoids after formalization. To increase toxoid purity, toxin purification was tried before formalization. Crude toxin was purified with ultrafiltration and ion-exchange chromatography. Purified toxin purity was improved 2.9 times higher than crude toxin, and purity was 2,560 Lf/mg PN. Purified toxin was detoxified with formalin and lysine, and potency test were performed. Toxoid, prepared from toxin treated with formalin and lysine, did not show reversion to toxin and purity was higher than the toxoid purified after formalization. Therefore, we concluded that the use of toxoid vaccine prepared from toxin purified is a useful method of minimize adverse reaction after injection of diphtheria vaccine.
본 연구에서는 디프테리아 toxin을 정제한 후 무독화하여 toxid를 생산하기 위하여 crude toxin을 2중 U.F를 통해 분자량 100.000이상의 단백질과 30.000이하의 배지 유래 단백질 및 색소를 제거한 결과 순도 1,300 Lf/mg PN의 toxin을 정제하였다. 이를 다시 DEAE-ion exehange chromatography를 통해 toxin을 정제한 후 무독화하여 순도 2,560 Lf/mg PN의 toxoid를 얻을 수 있었다. 이와 같이 생산된 디프테리아 toxoid는 동물 실험 결과 toxin으로 reversion이 발견되지 않았으며, 역가에 있어서도 crude toxin을 무독화한 후 정제한 toxoid와 비교하여 더 우수하였고 대한민국 생물학적 제제 기준에 규정된 성인용 디프테리아 백신 순도 기준 2,500 Lf/mg PN 이상에 적합하였다. 따라서 본 연구를 통해 성인용 디프테리아 백신의 생산 가능성을 확인하였다.