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目的利用简单的纯化工艺，获得具有生物活性的重组人神经生长因子（rhNGF）。方法利用PET－11d－NGF／BL21工程菌表达rhNGF，通过改变培养基成份优化发酵条件。建立了一种在提取包涵体后经一步强阳离子柱层析的简单、快速、重复性好的rhNGF纯化方法。纯化产物用鸡胚背根神经节伸长法测定其生物学活性。结果优化发酵条件后，表达量提高至约占总菌体的10．9％。经一步层析，重组蛋白的纯度可达到80％以上。1BU（生物活性单位）约为0．5μg／mL。结论包涵体复性表明，包涵体的复性与复性液中氧化型与还原型含疏基化合物的比例密切相关。抑制杂菌生长，可明显提高rhNGF的表达量。纯化的rhNGF为分析NGF结构与功能及探讨其临床应用提供了材料。
Objective To obtain bioactive recombinant human nerve growth factor (rhNGF) using a simple purification process. Methods The rhNGF was expressed by PET-11d-NGF / BL21 engineered bacteria and the fermentation conditions were optimized by changing the medium composition. A simple, rapid and reproducible purification method of rhNGF was established after one-step strong cation column extraction of inclusion bodies. The purified product was assayed for its biological activity by chick embryo dorsal root ganglia elongation method. Results After optimization of fermentation conditions, the expression level increased to about 10.9% of the total bacterial cells. After one-step chromatography, the purity of the recombinant protein can reach more than 80%. 1BU (bioactive units) is about 0.5 μg / mL. Conclusion Inclusion body refolding shows that the refolding of inclusion bodies is closely related to the ratio of oxidized and reduced forms of the so-called sparing group-containing compound in the refolding fluid. Inhibit the growth of bacteria, can significantly improve the expression of rhNGF. Purified rhNGF for the analysis of NGF structure and function and to explore the clinical application of materials.