Expression of recombinant rat Neurotrophin-3 in Chinese hamster ovary cells |
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| Authors: | Aimin Ji Siyun Shu Ming Li Xinmin Bao Hengqin Zou Zhongyi Zhang |
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| Affiliation: | 1. Zhujiang Hospital, The First Military Medical University, 510282, Guangzhou, China
2. Department of Tropical Disease, The First Military Medical University, 510515, Guangzhou, China
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| Abstract: | The CHO cell line stably producing recombinant rat NT-3 was established. The insertion of rNT-3 cDNA into transferred cell gonome was analyzed with Southern blot. The expressed protein was identified by Dot ELISA (enzyme-linked immunosorbent assay) and Western blot. Western blot showed a clear specific band of about 14 ku for NT-3. The mean level of rNT-3 in four NT-3cDNA/CHO cell lines was about 2 100 ng/106 cells/48 h determined by EIA. The conditioned-medium (CM) of NT-3cDNA/CHO cells could promote the fiber outgrowth of the dissociated dorsal root ganglion of 8-day-old chick embryos, which shows a dose-response relationship. A half-maximal concentration of the biological activity (EC50) of the recombinant protein was approximately 16.7 ng/mL. The MoAb 3W3 of NT-3 could neutralize the biological activity of the rNT-3. |
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