Department of Bioscience and Biotechnology, Sejong University, 98 Gunja-dong, Gwangjin-gu, Seoul, 143-747, Korea.
Cytotechnology. 2005 Jan;47(1-3):37-49. doi: 10.1007/s10616-005-3775-2.
Thirteen vitamins, twenty amino acids, hormones, inorganic salts, and other chemical agents, which constitute typical serum-free media, were evaluated for the development of fortified medium to enhance cell growth and productivity of recombinant antibody in the cultures of the recombinant Chinese hamster ovary (rCHO) cells. Two different rCHO cell lines, rCHO-A producing recombinant antibodies against the human platelet and rCHO-B secreting recombinant antibodies against the S surface antigen of Hepatitis B, respectively, were cultivated in batch suspension mode. Concentration of interested component in the tested medium was doubled to examine the fortification effect. Growth of rCHO-A cell and its antibody production were slightly improved with addition of either choline chloride, folic acid, thiamine[Symbol: see text]HCl, or Long(TM)R(3)IGF-I. On the other hand, in the cultivation of rCHO-B cell which was more sensitive to its environmental changes, hormones such as Long(TM)R(3)IGF-I and triiodothyronine (T(3)) as well as various vitamins involving choline chloride, i-inositol, niacinamide, pyridoxine HCl, and thiamine[Symbol: see text]HCl enhanced the cell growth and antibody production. Particularly, when concentration of consuming amino acid was doubled, remarkable increase in specific productivity was served, resulting in high final antibody concentration. These results were believed to provide a fundamental strategy of medium fortification useful for improvement of recombinant antibody production in serum-free medium.
十三种维生素、二十种氨基酸、激素、无机盐和其他化学物质构成了典型的无血清培养基，我们评估了这些物质，以开发强化培养基来提高重组中国仓鼠卵巢 (rCHO) 细胞培养物中细胞生长和重组抗体的生产力。分别用两种不同的 rCHO 细胞系 rCHO-A 和 rCHO-B 进行了分批悬浮培养，rCHO-A 细胞产生针对人血小板的重组抗体，rCHO-B 细胞分泌针对乙型肝炎表面抗原的重组抗体。在测试培养基中，感兴趣成分的浓度加倍，以检查强化效果。添加氯化胆碱、叶酸、硫胺素[Symbol: see text]HCl 或 Long(TM)R(3)IGF-I 可略微提高 rCHO-A 细胞的生长和抗体产量。另一方面，在对环境变化更为敏感的 rCHO-B 细胞培养中，Long(TM)R(3)IGF-I 和三碘甲状腺原氨酸 (T(3)) 等激素以及涉及氯化胆碱、肌醇、烟酰胺、盐酸吡哆醇和硫胺素[Symbol: see text]HCl 等多种维生素均能促进细胞生长和抗体产生。特别是当消耗氨基酸的浓度加倍时，特异性生产率显著提高，从而获得高的最终抗体浓度。这些结果为无血清培养基中提高重组抗体生产的培养基强化提供了一种基本策略。
