目的 对剪切力敏感哺乳动物细胞株进行灌流培养工艺开发与优化，以获得高蛋白产量。方法 利用N-1种子阶段灌流工艺获得高密度种子用于N阶段强化批次补料工艺开发；使用TPP摇管筛选灌流培养基及初步开发灌流工艺，再进行台式生物反应器工艺放大与优化。结果 “种子灌流+强化批次补料”混合工艺的单位体积产能相较于原始批次补料工艺提升46%；建立培养基筛选评分机制并筛选出1种混合培养基（基础+补料培养基）和1种通用灌流培养基，并用于2 L反应器规模灌流工艺放大，采用混合培养基和通用灌流培养基的灌流工艺单位体积产能分别提升335%和460%。结论 在N-1阶段和N阶段各自建立了剪切力敏感哺乳动物细胞株灌流工艺，并在N阶段获得高产量。

Objective  To develop and optimize the perfusion culture process for shear stress sensitive mammalian cell line to obtain high protein production. Methods  High-density seed cells were obtained by N-1 seed stage perfusion process for N stage intensified fed-batch process development. The perfusion medium suitable for the cell line was screened out and the process was further optimized at TPP tube scale. The scale-up and process optimization in benchtop bioreactors was then executed. Results  The unit volume capacity of the hybrid process of "seed perfusion process+intensified fed-batch process" was increased by 46% compared to original fed-batch process. Using the established medium rating system, one mixed medium (basal+feed medium) and one commercial perfusion medium were screened out and were scaled up to 2 L benchtop bioreactors. The mixed medium process increased the unit volume capacity by 335%, and the commercial perfusion medium process increased it by 460%. Conclusion  The perfusion culture processes for shear stress sensitive mammalian cell line at N-1 and N stages are established respectively, and designed for best production capacity at N stage.