b型流感嗜血杆菌培养及细菌溶解产物制备工艺研究

目的研究b型流感嗜血杆菌(Haemophilus influenza type b,Hib)培养条件及细菌溶解产物制备工艺。方法比较Hib在传统培养基和改良培养基的生长情况,优化改良培养基在生物反应器中对Hib培养条件(p H值、温度、溶解氧等),研究最佳菌体细胞破碎方法及细菌溶解产物纯化制备工艺。结果改良培养基可代替传统培养基用于Hib的生产培养,且Hib在p H7.4、温度为36℃、溶解氧为25%的改良培养基中,生长迅速,菌体产量最高。高压匀浆破碎法的破碎效果明显优于超声波破碎法,破碎效果达到98%以上,纯化后的细菌溶解产物的多糖含量、总氮含量、蛋白质含量、核酸含量及细菌内毒素含量均符合《中华人民共和国药典》(三部)2010版中b型流感嗜血杆菌结合疫苗的质量标准。结论初步建立了b型流感嗜血杆菌培养及细菌溶解产物制备工艺。

Bacterial lysate of Haemophilus culture and crushing process influenzae type b bacteria parameter optimization

Objective To explore the cultivation technology of Haemophihts influenzae type b and preparation technology in the bacterial lysate. Methods The growth status of Hib on the traditional media and modified media was compared. The culture conditions （ such as pI-I, temperature, dissolved oxygen, etc. ） for Hib bacterial cells were optimized. The best cell disruption method and the purified preparation process of cell lysates was studied. Results Improved medium can replace the traditional culture medium used for the production of Hib, and under the conditions of pH7.4 improved medium, 36 ℃ temperature and 25% of dissolved oxygen, Hib can rapid growth, and the highest yield can be obtained. Crushing effect of high pressure homogenizer crushing method was superior to sonication method, the crushing efficiency over 98%, bacterial lysates purified polysaccharide content, total nitrogen content, protein content, nucleic acid content and bacterial endotoxin levels were in line with＂ Pharmacopoeia of the People＇s Republic of China＂ （ three ） in the 2010 edition of Haemophilus influenzae type b vaccine combined with quality standards. Conclusion Initially established culture of Haemophilus influ- enzae type b bacteria dissolve the preparation process and the product.