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人神经生长因子的规模纯化
The Scale Purification of Human Nerve Growth Factor
【摘要】 建立一种简便、快捷的人神经生长因子(human Nerve Gtowth Factor, hNGF)的规模纯化方法,获得批量的hNGF。方法:以胎盘为原料,根据hNGF的性质,采用匀浆、超滤及一步层析法,规模纯化hNGF,并用SDS-PAGE及等电聚焦分别测定其分子量和等电点,利用鸡胚背根神经节法检测hNGF的生物活性。结果;纯化得单一组分的hNGF,测得其分子量约14.4 kD,等电点约为9.3,比活约1.5×10~4U/mg。纯化产物与抗重组人神经生长因子抗体有很强的免疫交叉反应。结论:该纯化工艺可得到批量、高纯度的人神经生长因子。
【Abstract】 Purpose: The aim was to establish a simple protocol for the purfication of the human nerve growth factor (hNGF) from human term placentas on large scale to obtain enough hNGF. Methods: The purification procedure included tissue homogenization, ultrafiltration and single CM-cellulose column chromatography. The molecular weight (MW) and isoelectric point (PI) of the purity was measured by means of SDS-PAGE and isoelectric focus electrophoresis, respectively. The bioactivity was assayed by the method of dorsal root ganglion (DRG). Results: A homogenous product of MW 14. 4 kD, pI about 9. 3 was obstained through this protocol. The activity of this product was about 1. 5 × 10~4 U/mg. The purity could interact with the monoclonal antibody against recombinant human NGF (rhNGF). Conclusion: The results indicated that we could get hNGF in batch process through the simple protocol reported here.
- 【文献出处】 中国生化药物杂志 ,CHINESE JOURNAL OF BIOCHEMICAL PHARMACEUTICS , 编辑部邮箱 ,1999年03期
- 【分类号】R915
- 【被引频次】5
- 【下载频次】108