全人源抗VEGF165单克隆抗体培养工艺的研究

为研究一种表达全人源抗VEGF165单克隆抗体(HVAB)的重组DG44细胞的培养工艺。在摇瓶培养阶段，利用常见的流加和分阶段温度控制培养的方法，初步确定培养工艺。在生物反应器培养阶段，研究了不同pH范围对DG44细胞生长和单克隆抗体表达的影响。结果表明，通过补料可使HVAB表达量从101 mg/L提高到654 mg/L；通过在生长中期的适当降温，使细胞终活性保持在80%以上；pH控制范围6.4~7.4更适合DG44细胞的生长和HVAB表达。反应器中的抗体表达量为526 mg/L，与对照摇瓶相比降低了11%，为之后的中试研究奠定了工艺基础。
This study was focus on the optimizing cell culture process of recombinant DG44 cells which expressing novel fully human anti-VEGF165 monoclonal antibody(HVAB). Feed-batch culture and two-phase temperature control were studied for optimizing the HVAB productivity in shaken flasks. The influences of pH changes were determined to study the growth of DG44 cell and expression of HVAB in bioreactors. The HVAB productivity was rose from 101 mg/L to 654 mg/L after feed-batch culture in shaken flask, and cell viability maintained above 80% after reduce the temperature in middle growth phase. It is also suggested that pH range at 6. 4-7. 4 is beneficial to DG44 cells growth and HVAB expression. The productivity in bioreactor is 526 mg/L decreased 11% compare with in shaken flask, which laid an foundation for further pilot scale production