论文部分内容阅读
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 specifie band of about 14 ku for NT-3. The mean level of rNT-3 in four NT-3eDNA/CHO cell lines was about 2 100 ng/10~6 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.
The insertion of rNT-3 cDNA into transferred cells gonome was analyzed with Southern blot. The expressed protein was identified by Dot ELISA (enzyme-linked immunosorbent assay) and Western blot. The mean level of rNT-3 in four NT-3 cDNA / CHO cell lines was about 2 100 ng / 10 ~ 6 cells / 48 h determined by EIA. The Western blot showed a clear specifi band of about 14 ku for NT-3. conditioned-medium (CM) of NT-3 cDNA / 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.