工艺研发

风疹病毒BRDⅡ减毒株E1区连续传代稳定性研究

  • 简哲 ,
  • 李欣娜 ,
  • 刘爽 ,
  • 杨天石 ,
  • 李杨 ,
  • 孙雅 ,
  • 王刚 ,
  • 邱梅
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  • 北京生物制品研究所有限责任公司疫苗一室,北京 100176

网络出版日期: 2025-08-16

Genetic stability of rubella virus BRDⅡ attenuated strain after successive passage

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  • Vaccine Department One, Beijing Institute of Biological Products Co., Ltd., Beijing 100176, China

Online published: 2025-08-16

摘要

目的 评估风疹病毒BRDⅡ减毒株(BRDⅡ株)E1区在人二倍体细胞2BS株(2BS细胞)中的连续传代稳定性。方法 将BRDⅡ株28代病毒在2BS细胞中连续传10代分别获得第29—38代病毒,依据中国药典2020年版三部对不同代次病毒进行生物学特性检测,将28代病毒(主种子批)、31代病毒(工作种子批)以及38代病毒进行全基因测序后对其核苷酸序列进行分析比对。结果 BRDⅡ株28代病毒在2BS细胞中连续传10代,其细胞病变程度基本稳定。基因测序发现,结构蛋白E1区第8281nt位点共出现1个突变,为同义突变。结论 BRDⅡ株在2BS细胞中自28代连续传至38代,病毒蛋白主要功能区E1区的氨基酸序列高度保守,具有良好的传代稳定性。

本文引用格式

简哲 , 李欣娜 , 刘爽 , 杨天石 , 李杨 , 孙雅 , 王刚 , 邱梅 . 风疹病毒BRDⅡ减毒株E1区连续传代稳定性研究[J]. 国际生物制品学杂志, 2024 , 47(2) : 67 -71 . DOI: 10.3760/cma.j.cn311962-20230330-00029

Abstract

Objective To evaluate the genetic stability of rubella virus BRDⅡ attenuated strain(BRDⅡ strain) E1 region after successive passage in human diploid cell line 2BS strain(2BS cell) . Methods The 28th generation virus of BRDⅡ strain was subcultured in 2BS cell for 10 generations to obtain the 29th-38th generation viruses. The biological characteristics of different generation viruses were detected according to Chinese pharmacopoeia 2020 edition volume Ⅲ. The nucleotide sequences of 28th generation virus (master seed lot), 31st generation virus (working seed lot) and 38th generation virus were analyzed and compared. Results The 28th generation virus of BRDⅡ strain was continuously transmitted for 10 generations in 2BS cell, and the cytopathic effect was generally stable. Gene sequencing revealed only one synonymous mutation occurred at 8281nt of E1 region of the structural protein. Conclusion After subcultured from 28th to 38th generations in the 2BS cell,the amino acid sequence of BRDⅡ strain E1 region, the main functional region of the viral protein, is highly conserved and has good passaged stability.
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