人神经生长因子的规模纯化

The Scale Purification of Human Nerve Growth Factor

目的：建立一种简便，快捷的人神经生长因子的规模纯化方法，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。

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.