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目的 :检测人 β NGF基因转染CHO细胞后培养上清中所表达的 β NGF的生物学活性及分离纯化。 方法 :采用脂质体介导的真核细胞转染 ,运用形态学观察和MTT法进行检测做定性定量分析 ,蛋白观察用SDS PAGE电泳。结果 :转染 β NGF基因的CHO细胞可分泌能促进PC12细胞生长的活性 β NGF ,且可透过血脑屏障。 结论 :转染 β NGF基因的CHO细胞所分泌的 β NGF ,接近自然合成的蛋白 ,具有较高生物学活性 ,为NGF的生物制药奠定了实验基础。
OBJECTIVE: To detect the biological activity of β NGF expressed in culture supernatant after transfecting human β NGF gene into CHO cells and to isolate and purify it. Methods: Liposome-mediated eukaryotic cell transfection was used for qualitative and quantitative analysis by morphological observation and MTT assay. The protein was observed by SDS PAGE electrophoresis. Results: CHO cells transfected with β NGF gene secreted active β NGF, which can promote the growth of PC12 cells and could cross the blood-brain barrier. CONCLUSION: The β NGF secreted by CHO cells transfected with β-NGF gene is close to the naturally-synthesized protein, which has high biological activity and lays the foundation for biopharmaceutical production of NGF.