AI Article Synopsis
- A CHO cell line was created to produce recombinant rat NT-3, and the integration of rNT-3 cDNA was confirmed using Southern blot analysis.
- The presence of the NT-3 protein was verified through Dot ELISA and Western blot, where a specific band around 14 kDa was observed.
- The conditioned medium from these cells promoted the growth of nerve fibers in chick embryo dorsal root ganglia, with a half-maximal effective concentration (EC50) of 16.7 ng/mL, and the MoAb 3W3 could neutralize this activity.
Article 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/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.
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| http://dx.doi.org/10.1007/BF02881584 | DOI Listing |
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