CHO细胞糖基化同质性无血清培养基的研发及机制

无血清培养基（serum-free medium, SFM）是哺乳动物细胞表达系统生产重组蛋白/抗体治疗药物的必需原料。近70%的重组蛋白/抗体药物是由中国仓鼠卵巢（Chinese hamster ovary, CHO）细胞产生的。重组蛋白药物的糖基化异质性严重影响其功能和活性，且机制不清楚。研究发现培养基成分会影响重组蛋白的糖基化异质性。申请者前期研发了适合CHO细胞的具有高密度、高活性和高表达量的SFM。本项目拟在前期基础上，研发用于重组蛋白生产的糖基化同质性SFM，并探讨其分子机制。通过构建重组蛋白（抗体）稳定表达细胞株，筛选影响重组蛋白糖基化的关键组分，分析其对重组蛋白的糖基化同质性的作用。进一步采用代谢组学和RNA组学挖掘影响糖基化异质性的关键因子。该项目完成有望提供一种高值国产无血清培养基，提升高端重组蛋白药物的国产自主研发水平。

Serum-free medium (SFM) is essential for recombinant therapeutic protein or antibody produced by the mammalian expression system. Today, nearly 70% of the recombinant therapeutic proteins or antibodies are produced in Chinese hamster ovary (CHO) cells. Glycosylation heterogeneity of recombinant therapeutic proteins seriously affects their function and activity, and the mechanism is unclear. It was found that the composition of culture medium can affect the glycosylation heterogeneity of recombinant protein. In the previous study, we have developed SFM with high density, high activity, and high expression for CHO cells. This project intends to develop SFM with homogeneous glycosylation for the production of recombinant protein and explore its molecular mechanism. Through the construction of stable cell lines with expressing recombinant protein or antibody, the key components affecting the glycosylation of recombinant protein will be screened, and their effects on the glycosylation homogeneity of recombinant protein will be analyzed. Metabonomics and RNAmics will be used to explore the key factors affecting glycosylation heterogeneity. The completion of the project is expected to provide a high-value domestic SFM and improve the domestic independent R & D level of high-end recombinant therapeutic proteins.