对rhEndostatin包涵体蛋白的色谱复性条件进行研究,摸索出最佳的复性、纯化条件,解决其在大肠杆菌中表达提取时遇到的蛋白变性问题。按实验条件对表达的rhEndostatin包涵体蛋白进行提取、洗涤、变性;利用正交法设计重组蛋白的色谱复性、纯化实验;利用SP-Sepharose Fast Flow阳离子交换柱对rhEndostatin包涵体蛋白进行了色谱复性、纯化。最佳复性条件为rhEndostatin上样浓度为2.5 mg/mL,洗脱体积流量为1/20床体积/min(3.5 mL/min),NaCl浓度0.1 mol/L,GSH/GSSG的浓度比为3/0.3 mmol/L。在此条件下纯度约97%,蛋白回收率为90%,重组蛋白的活性最高,IC50达到5μg/mL。 The chromatographic refolding conditions of rhEndostatin inclusion body protein were explored to find out the best refolding and purification conditions and to solve the protein denaturation problems encountered in the expression and extraction of rhEndostatin. The expressed rhEndostatin inclusion body protein was extracted, washed and denatured according to the experimental conditions. The chromatographic refolding and purification experiments of the recombinant protein were designed by orthogonal design. The protein of rhEndostatin inclusion body was chromatographed by SP-Sepharose Fast Flow cation exchange column Sexual, purified. The optimum refolding conditions were rhEndostatin loading concentration 2.5 mg / mL, elution volume flow rate 1/20 bed volume / min (3.5 mL / min), NaCl concentration 0.1 mol / L and GSH / GSSG concentration ratio 3 /0.3 mmol / L. Under these conditions, the purity was about 97%, the protein recovery was 90%, and the recombinant protein had the highest activity with an IC50 of 5μg / mL.