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Effects of Supplementation of Various Medium Components on Chinese Hamster Ovary Cell Cultures Producing Recombinant Antibody

机译:补充各种培养基成分对中国仓鼠卵巢细胞产生重组抗体的影响

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摘要

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⋅HCl, or LongTMR3IGF-I. On the other hand, in the cultivation of rCHO-B cell which was more sensitive to its environmental changes, hormones such as LongTMR3IGF-I and triiodothyronine (T3) as well as various vitamins involving choline chloride, i-inositol, niacinamide, pyridoxine HCl, and thiamine⋅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.
机译:评估了构成典型无血清培养基的13种维生素,20种氨基酸,激素,无机盐和其他化学试剂的强化培养基的开发,以增强重组仓鼠培养物中细胞的生长和重组抗体的生产力卵巢(rCHO)细胞。以分批悬浮方式培养两种不同的rCHO细胞系,分别是产生抗人血小板的重组抗体的rCHO-A和分泌针对乙型肝炎S表面抗原的分泌rCHO-B的重组抗体。测试介质中目标组分的浓度加倍,以检查强化效果。加入氯化胆碱,叶酸,硫胺素·HCl或Long TM R 3 IGF-I可稍微改善rCHO-A细胞的生长及其抗体的产生。另一方面,在对环境变化更敏感的rCHO-B细胞的培养中,Long TM R 3 IGF-I和三碘甲状腺素(T3 )以及各种维生素,包括氯化胆碱,异肌醇,烟酰胺,吡ido醇HCl和硫胺素HCl,可增强细胞生长和抗体产生。特别地,当消耗氨基酸的浓度加倍时,可以显着提高比生产率,从而导致高的最终抗体浓度。认为这些结果提供了用于增强无血清培养基中重组抗体产生的培养基强化的基本策略。

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