基于单纯型设计和部件搜索方法的CHO细胞无血清培养基的高通量优化

High Through-Put Screening and Optimization of Serum-free Media Development for CHO Cells Using Simplex Lattice and Elements Dialogue Design Methods

氨基酸的浓度与配比对细胞生长和产物表达有重要作用。在自主研发无血清培养基的基础上,通过单纯型格子实验设计优化氨基酸浓度及配比后发现,优化的无血清培养基明显促进了细胞生长。生物反应器批式培养中,最高活细胞密度达52.6×105 cells·mL-1,抗体融合蛋白产量52.2 mg·L-1,相对于商业培养基Ex-cell 302,分别提高了26.7%和11.5%。在此基础上,通过部件搜索的方法确认具有显著正影响的氨基酸为谷氨酰胺、脯氨酸和甲硫氨酸。基于上述研究工作,在2 L反应器采用自主研发的优化无血清培养基,以葡萄糖为关键控制参数,进行两阶段动态流加培养过程,CHO细胞最大活细胞密度达94.6×105 cells·mL-1,抗体融合蛋白终产量600 mg·L-1,相对于商业培养基Ex-cell 302批式培养,分别提高了2.75倍和11.8倍。本研究工作为抗体融合蛋白的高效工业化生产奠定了基础。

Based on statistical analysis, cell growth was optimization of the concentration and proportion of amino design methods. Using the optimized serum-free media, maximum antibody concentration are 52.6×10^5 cells.mL^ 1 improved in optimized serum-free media via the acids using simplex lattice and elements dialogue the maximum viable cell density （MVCD） and the and 52.2 mg.L^1, respectively, and these values are increased by 26.7% and 11.5% respectively comparing to those obtained by using the commercial medium Ex-cell302. Glutamine, proline and methiorline are confirmed to have positive effects through the elements dialogue experiment. Moreover, two-stage dynamic fed-batch culture for CHO cells was studied in a 2 L bioreactor. The feeding strategy was based on using the glucose concentration as the key controlling parameter. The maximum viable cell density reaches to 94.6×10^5 cells.mL^11, and 600 mg.L^- 1 antibody concentration was achieved within 10 days of cultivation. The maximum cell concentration and antibody production are improved by 2.75 and 11.8 times comparing to the results obtained by commercial medium cultivation. This research is useful for efficient industrial production of antibody fusion proteins.