摘要
目的评估自主研发培养基QS作为首选培养基用于冻干人用狂犬病疫苗(鸡胚成纤维细胞)生产过程的可行性。方法分别制备基于自主研发培养基QS和其他3种商业化培养基(X1、X2、X3)的鸡胚成纤维细胞悬液,观察和比较细胞形态和生长特性;以Flury HEP株接种鸡胚成纤维细胞(MOI=0.003),分别通过直接免疫荧光法、蛋白质印迹法(Western blotting)和酶联免疫吸附测定(enzyme-linked immunosorbent assay,ELISA)比较不同培养基条件下收获液中病毒滴度和G蛋白含量。结果在细胞浓度1×10~6个/mL条件下,4种培养基培养的鸡胚成纤维细胞在形态上均无显著区别;但是自主研发培养基QS和X2的病毒收获液的G蛋白含量在第4天时分别为0.66 IU/mL和0.63 IU/mL,高于X1的0.5 IU/mL和X3的0.3 IU/mL。在第6天时分别为0.92 IU/mL和0.88 IU/mL,高于X1的0.64 IU/mL和X3的0.52 IU/mL,说明基于自主研发培养基QS和X2的病毒收获液在G蛋白含量方面具有明显优势。结论自主研发培养基QS可以用于冻干人用狂犬病疫苗(鸡胚成纤维细胞)的生产。
Objective To evaluate the feasibility for a self-developed medium(QS)in production of rabies vaccine(chicken embryo fiboblast)for human use.Methods Chicken embryonic fibroblasts suspensions were prepared by using QS and three other commercial mediums(X1,X2,X3),respectively,morphology and growth characteristics of cultured cells were observed and compared.Chicken embryonic fibroblasts were inoculated with Flury HEP strain(MOI=0.003).Virus titer and G protein content of harvested suspension based on different mediums were tested by direct immunofluorescence,Western blotting and ELISA.Results There was no an apparent difference in morphology of chicken embryonic fibroblasts cultured in four mediums at the concentration of 1×10~6 cells/mL;however,the content of G protein in harvested virus suspension of QS and X2 was 0.66 IU/mL and 0.63 IU/mL respectively on the 4 th day,which was higher than 0.50 IU/mL of X1 and 0.30 IU/mL of X3.Followed by reached to 0.92 IU/mL and 0.88 IU/mL,respectively on the 6 th day,higher than 0.64 IU/mL of X1 and 0.52 IU/mL of X3,so the harvested virus suspension had obvious advantages in G protein content based on QS and X2.Conclusion The self-developed medium QS could be used in production of freeze-dried rabies vaccine(chicken embryo fibroblasts)for human use.
作者
陈磊
苏波
牛彪
阮俊程
郭跃
王德慧
顾玉林
田大勇
CHEN Lei;SU Bo;NIU Biao;RUAN Jun-cheng;GUO Yue;WANG De-hui;GU Yu-lin;TIAN Da-yong(R&D Department,Shanghai King-cell Biotechnology Co.,Ltd.,Shanghai 201506,China)
出处
《微生物学免疫学进展》
2020年第1期56-62,共7页
Progress In Microbiology and Immunology
基金
国家自然科学基金(31702248).
作者简介
陈磊(1988-),男,助理工程师,主要从事病毒性疫苗的研发工作;通信作者:田大勇,工程师,E-mail:tiandayong@king-cell.com
