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目的 :测定NT -3cDNA/CHO细胞表达的基因重组NT -3的生物学活性。方法 :体外培养 8日龄鸡胚背根神经节 (DRG) ,加入条件培养基 ,测量背根节神经突起生长的长度。结果 :NT -3cDNA/CHO细胞培养 4 8h后的无血清条件培养基 1∶2 4稀释后即能促进DRG神经突起的生长 ;随着其中NT -3浓度增加 ,神经突起长度也逐渐延长 ,有明显的量效关系 ;DRG培养 4 8h长出的神经突起与 2 4h相比 ,明显长而致密 ;NT -3生物活性的EC50 值为1 6.7ng/ml;NT -3浓度为 1 2 5ng/ml时 ,神经突起的长度达到最大值 ;NT -3cDNA/CHO细胞培养 72h的条件培养基中NT -3的活性高于培养 4 8h的条件培养基 ;NT -3单克隆抗体 3W 3的浓度为 0 .5μg/ml即能抑制NT -3的生物活性 ;抗体浓度为 1 μg/ml时 ,NT -3的生物活性降低一半。 结论 :基因重组NT -3有较高的生物活性
OBJECTIVE: To determine the biological activity of recombinant NT-3 expressed by NT-3 cDNA / CHO cells. Methods: 8-day old chick embryo dorsal root ganglion (DRG) was cultured in vitro and conditioned medium was added to measure the length of dorsal root ganglion neurite. Results: NT-3 cDNA / CHO cells cultured in serum-free condition for 48h could promote the growth of DRG neurite after 1: 2 dilution. With the increase of NT-3 concentration, the length of neurite was also prolonged Significant dose-effect relationship; DRG cultured 48 h neurites grew significantly longer and dense compared with 2 4 h; NT 3 biological activity of EC50 value of 1 6.7ng / ml; NT -3 concentration of 125ng / NT-3cDNA / CHO cells cultured for 72h had higher NT-3 activity than that of conditioned medium incubated for 48h. The concentration of NT-3 monoclonal antibody 3W 3 was 0 .5μg / ml that can inhibit the biological activity of NT -3; antibody concentration of 1 μg / ml, NT -3 bioactivity reduced by half. Conclusion: Gene recombinant NT -3 has high biological activity