人体胎盘神经生长因子的纯化和鉴定

Purification and Characterization of Nerve Growth Factor of Human Placenta Origin

采用人胎盘为原料，利用离心、超滤和SephadexG-100、DEAE－SephadexA－50、SephadexG－150层析等技术，在中性条件下，分离出7S神经生长因子（7SNGF）；再在酸性条件下，利用柱状等电聚焦电泳法分离出活性亚单位-β-NGF，利用鸡胚背根神经节培养法直接测定神经生长因子的生物活性，其比活性为8000u／mg；用十二烷基磺酸钠不连续聚丙烯酰胺凝胶电泳（SDS－PAGE）测得β-NGF分子量为13KD；高效液相PEK－125柱层析法得单一洗脱峰证明为单一成分。

7S nerve growth factor (7SNGF) was seperated from human placenta by centrifugation, ultrafiltration, Sephadex G^100, DEAE-Sephadex A-50 and Sephadex G150 chromatography in neutral condition. Then β-NGF was isolated by column isoelectrofocusing electrophoresis in acidic condition. The biologlcal activity of 7S NGF was directly determined by chick embryo dorsal root ganglion cuItivation. The result showed that the speclfic activity of 7S NGF was 8000U/mg. Identified by high-performance liquid chromatography (HPLC) with PEK-125 column, only one peak appeared. It indicated that the 7S NGF was only a single component. The molecular weight of β-NGF detected by SDSPAGE was 13 KD.