CD-1, a genetically-engineered CHO cell line, was cultivated with a Biosilon microcarrier culture system.We successfully cultivated CD-1 cells to a very high density (over1×107cells/ml). Prourokinase was stably s...CD-1, a genetically-engineered CHO cell line, was cultivated with a Biosilon microcarrier culture system.We successfully cultivated CD-1 cells to a very high density (over1×107cells/ml). Prourokinase was stably secreted at about 180 IU/ 1e6 cells/24 h. Experiments showed that CD-1 cells growing on Biosilon microcarriers were able to spontaneously release from the microcarriers, then reatthch and proliferate on fresh microcarriers. This makes it very easy to scale up preduction. The microcarriers could be reused several times without affecting adhesion. proliferation and prourokinase secretion. With CMPECC membrane radial flow chromatography and MPG chromatography, the prourokinase in conditioned medium could be purified to a specific activity of 1×105 IU/mg of protein. The purification factor was about 600 fold, and approxiamately 90 % of the biological activity was recovered.展开更多
rβ-13 CHO 工程细胞在 MC-1型微载体上可良好地附着和增殖,当载体浓度为10mg/ml 时,细胞密度可高达4.5×10~6细胞/ml。由于该细胞具有可自动从载体解离,并再附着和增殖于新载体表面的特性,故采用直接加入新鲜载体和培养基的方法,...rβ-13 CHO 工程细胞在 MC-1型微载体上可良好地附着和增殖,当载体浓度为10mg/ml 时,细胞密度可高达4.5×10~6细胞/ml。由于该细胞具有可自动从载体解离,并再附着和增殖于新载体表面的特性,故采用直接加入新鲜载体和培养基的方法,即可方便地扩大生产。这些结果表明用微载体悬浮培养系统生产 CHO 工程细胞产品将是可行的。展开更多
文摘CD-1, a genetically-engineered CHO cell line, was cultivated with a Biosilon microcarrier culture system.We successfully cultivated CD-1 cells to a very high density (over1×107cells/ml). Prourokinase was stably secreted at about 180 IU/ 1e6 cells/24 h. Experiments showed that CD-1 cells growing on Biosilon microcarriers were able to spontaneously release from the microcarriers, then reatthch and proliferate on fresh microcarriers. This makes it very easy to scale up preduction. The microcarriers could be reused several times without affecting adhesion. proliferation and prourokinase secretion. With CMPECC membrane radial flow chromatography and MPG chromatography, the prourokinase in conditioned medium could be purified to a specific activity of 1×105 IU/mg of protein. The purification factor was about 600 fold, and approxiamately 90 % of the biological activity was recovered.
文摘rβ-13 CHO 工程细胞在 MC-1型微载体上可良好地附着和增殖,当载体浓度为10mg/ml 时,细胞密度可高达4.5×10~6细胞/ml。由于该细胞具有可自动从载体解离,并再附着和增殖于新载体表面的特性,故采用直接加入新鲜载体和培养基的方法,即可方便地扩大生产。这些结果表明用微载体悬浮培养系统生产 CHO 工程细胞产品将是可行的。
