高密度接种强化CHO细胞培养工艺的研究  

Study on Strengthen CHO Cell Culture Technology by High Density Inoculation

作　　者：赵丽丽 朱玉强 徐凯 朱中松 张贵民 刘忠 ZHAO Lili;ZHU Yuqiang;XU Kai;ZHU Zhongsong;ZHANG Guimin;LIU Zhong(Lunan New Time Bio-tech Co.,Ltd.,Linyi 273400;State Engineering Lab.of High Expression of Mammalian Cells,Linyi 273400;Lunan Pharmaceutical Group Co.,Ltd.,Linyi 276000)

机构地区：[1]鲁南新时代生物技术有限公司,山东临沂273400 [2]哺乳动物细胞高效表达国家工程实验室,山东临沂273400 [3]鲁南制药集团股份有限公司,山东临沂276000

出　　处：《中国医药工业杂志》2024年第11期1511-1518,共8页Chinese Journal of Pharmaceuticals

基　　金：泰山产业领军人才工程项目(tscy20200329、tscx202306088)。

摘　　要：为确定不同高密度接种对双特异性抗体表达量及质量的影响、强化双特异性抗体细胞培养工艺,分别以低、中、高密度[每1 mL分别含(10±1)×10^(6)、(15±1)×10^(6)和(20±1)×10^(6)个细胞]将双特异性抗体细胞接种于3 L生物反应器,补料分批次培养,并评估不同接种密度对抗体表达量及质量的影响。结果表明,与较低的高密度接种相比,中等及较高的高密度接种强化工艺所得的抗体表达量提高至1.43 g/L,且抗体质量无显著差异。相同培养周期(16 d)内,在抗体质量不受影响和节约成本的前提下,接种密度应采用中等高密度,抗体表达量可提高到1.43 g/L,与常规工艺(0.79 g/L)相比提高了81%。随后将中等高密度接种强化工艺成功放大至30 L生物反应器,为后续抗体工艺开发提供了宝贵的经验。To determine the influence of different high inoculation densities on the expression level and quality of bispecific antibodies and strengthen the cell culture process,bispecific antibody cells were inoculated into a 3 L bioreactor at low,medium and high grades of high inoculation density[containing(10±1)×10^(6),(15±1)×10^(6)and(20±1)×10^(6)cells per 1 mL,respectively]for fed-batch culture.The influences of different inoculation densities on antibody expression level and quality were evaluated.The results showed that compared with low grade of high inoculation density,the antibody expression levels of the intensified medium or high grade of high inoculation density process increased to 1.43 g/L,and there was no significant difference in antibody quality.Within the same culture period(16 d),on the premise of no influence on antibody quality and reduction of costs,medium grade of high inoculation density should be adopted for inoculation density,and the antibody expression level could be increased to 1.43 g/L,which was an 81%increase compared with the conventional process(0.79 g/L).Subsequently,the intensified medium grade of high inoculation density process was successfully scaled up to a 30 L bioreactor,providing valuable experience for subsequent antibody process development.

关 键 词：双特异性抗体 培养工艺 补料批次培养 放大培养 

分 类 号：TQ464.7[化学工程—制药化工]