广西眼镜蛇毒神经生长因子分离纯化方法

Separation and Purification of Nerve Growth Factor from Guangxi Cobra Venom

为了得到更高纯度和活性的广西眼镜蛇毒神经生长因子(never growth factor,NGF),我们对原有的分离纯化方法进行改进。采用DEAE CL-6B离子交换柱、Sephadex G-50凝胶层析柱、Macro-prep High S离子交换柱结合的方法进行分离。在分离纯化过程中,采用PC12细胞检验每一步所得结果中的NGF活性,并测定NGF诱导PC12细胞分化的最小浓度。经DEAE CL-6B离子交换柱、Sephadex G-50凝胶层析柱、Macro-prep High S离子交换柱分离纯化后,得到的第二峰具有NGF活性,并且已达到电泳纯。经PC12细胞验证后,确定NGF诱导PC12细胞分化的最小浓度为0.1μg/m L。

In order to get the high activity and pure nerve growth factor（NGF） of Guangxi cobra venom, we improved the method of separation and purification. It was separated and purified by DEAE CL-6B cation-exchange column, Sephadex G-50 gel filtration and Macro-prep High S cation-exchange column. In the results of each step in the process of separating and purifying, we used PC12 cells to assay the NGF activity, and determined the minimal concentration of NGF to induced the differentiation of PC12 cells. After separating and purifying by DEAE CL-6B cation-exchange column, Sephadex G-50 gel filtration and Macro-prep High S cation-exchange column, we got the second peak with NGF activity, and it achieved electrophoresis purity. After verifying, the minimal concentration of NGF to induced the differentiation of PC12 cells was 0.1 μg/mL.