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Mol Biote:周保罗等开发出人单克隆抗体新型制备技术

  1. Mol Biote
  2. 单克隆抗体
  3. 果蝇

来源:上海巴斯德所 2012-11-18 13:15

近日,国际著名杂志Molecular Biotechnology 在线刊登了上海巴斯德研究所周保罗研究组的最新研究成果“High Yield of Human Monoclonal Antibody Produced by Stably Transfected Drosophila Schneider 2 Cells in Perfusion Culture Using Wave Bioreact

近日,国际著名杂志Molecular Biotechnology 在线刊登了上海巴斯德研究所周保罗研究组的最新研究成果“High Yield of Human Monoclonal Antibody Produced by Stably Transfected Drosophila Schneider 2 Cells in Perfusion Culture Using Wave Bioreactor,”这是首次关于用波浪生物反应器灌注培养稳转了人单克隆抗体基因的果蝇S2细胞来产生人单克隆抗体的报道。

治疗性人单克隆抗体已经增长为数十亿美元的产业,因而建立一个稳定的培养条件以满足人单克隆抗体的大规模表达是非常重要的。波浪生物反应器于二十世纪九十年代后期被用于哺乳动物及昆虫细胞的蛋白表达,但是用波浪生物反应器来灌注培养稳转了人单克隆抗体基因的果蝇稳转细胞表达人单克隆抗体蛋白尚属首次。

在该研究中上海巴斯德所的研究助理王璐岚和博士生胡红星在周保罗教授的指导下克隆了抗高致病性禽流感病毒的血凝蛋白的人单克隆抗体基因,并构建到果蝇诱导型表达载体里面,稳转了果蝇S2细胞系,通过有限稀释法拿到高表达人单克隆抗体的单细胞克隆。对稳转了人单克隆抗体的单克隆细胞株分别用来在灌注和非灌注的生物反应器中进行比较培养,发现灌注培养方法在获得细胞数量与表达抗体蛋白产量上明显优于后者,且这两种方法生产的抗体都是有中和活性的。这些结果表明用波浪生物反应器进行灌注培养S2稳转细胞系进行大规模蛋白表达是非常好的方法。

该研究是与GE公司研发部Fast Trak中心的杨建军博士合作完成的,得到了国家自然科学基金及李嘉诚基金会的资助。(生物谷Bioon.com)

High Yield of Human Monoclonal Antibody Produced by Stably Transfected Drosophila Schneider 2 Cells in Perfusion Culture Using Wave Bioreactor

Lulan Wang, Hongxing Hu, Jianjun Yang, Feng Wang, Christian Kaisermayer and Paul Zhou

Since it was first introduced in late 1990s Wave bioreactor has been used for protein production by mammalian and insect cell lines. However, using Wave bioreactor to produce human monoclonal antibody by stable Drosophila Schneider 2 (S2) cell transfectants has not been reported before. In this study, S2 cells were co-transfected with an inducible vector expressing human monoclonal antibody heavy and light chains, respectively, specific for hemagglutinin (HA) of H5N1 influenza virus. Stable S2 transfectant clone was selected by limiting dilution assay. Stable S2 transfectant clone that produce the highest amount of human monoclonal antibody was inoculated into two 2-l disposable cellbags, where cell growth and antibody production were compared between batch and perfusion cultures using Wave bioreactor. Here, we report that maximum viable cell density reached 1.06 × 107 cells/ml in batch culture; whereas 1.04 × 108 cells/ml was achieved in perfusion culture. The maximum volumetric antibody productivity in batch culture was 52 mg/l/day; while perfusion culture yielded 1,437 mg/l/day. As a result, the total antibody production was 201 mg in batch culture and 8,212 mg in perfusion culture. The antibody produced by both cultures displays full neutralizing activity. Thus, our results provide strong support for using Wave bioreactor in perfusion culture for a large-scale production of human monoclonal antibody by stable S2 cell transfectants.

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