一种重组人表皮生长因子的纯化工艺研究

A Preparation Method of Recombinant Human Epidermal Growth Factor

目的:以毕赤酵母细胞通过基因重组技术表达重组人表皮生长因子(Human Epidernal Growth Factor,以下简称h EGF)发酵液为原料,建立一种h EGF纯化方法。方法:以高速离心技术、膜过滤技术,浓缩脱盐技术为主的粗纯工艺;应用弱阳离子交换层析和凝胶色谱层析为主要精细纯化工艺,h EGF样品最终冻干保存。结果:以该纯化工艺最终可获得高纯度的h EGF,经Tricine-SDS-PAGE电泳检测及HPLC-C8色谱柱分析检测其纯度可达92%以上。结论:应用此纯化工艺得到的h EGF纯度大于92%、在化妆品原料方面可用于进一步工业化研究。

Objective： To establish a method of purifying recombinant Human Epidermal Growth Factor（hereinafter as h EGF）, which takes fermented liquid as raw materials derived from Pichia Cell expression via gene recombination technology. Methods： Using the high-speed centrifugal, membrane filtration and concentrated desalination technology as the major coarse purification methods. Ion exchange chromatography and Gel chromatography were applied as the primary fine purification method. The h EGF sample was freeze-dried for final preservation. Results： Using this method, The h EGF can be obtained with high purity. With Tricine-SDS-PAGE and HPLC-C8 column analysis, the purity could reach 92% and even higher. Conclusion： The purity of h EGF is higher than 92% by applying this purification method, which also could be used for further mass production in cosmetics raw material field.