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目的 :研究血管抑素 (angiostatin)的临床应用价值 ,将其开发为多肽类药物。方法 :以RT PCR从胎儿肝脏钓取人血管抑素全编码区cDNA ,使用Pichia分泌型酵母表达系统表达重组人血管抑素 ,重组蛋白经肝素亲和层析纯化后 ,以鸡胚尿囊膜法血管生成实验及小鼠创伤愈合实验检测活性。结果 :重组人血管抑素在酵母系统中得到较高量表达 (5mg/L) ,经肝素亲和层析纯化 ,SDS PAGE显示分子量为 43kD。活性实验表明 ,重组蛋白能够抑制新生血管形成、抑制伤口愈合。结论 :重组人血管抑素在酵母系统中实现高效表达 ,并具有很好的生物学活性。
Objective: To study the clinical value of angiostatin and to develop it as a peptide drug. Methods: cDNA of full-length human angiostatin cDNA was isolated from fetal liver by RT PCR. Recombinant human angiostatin was expressed by Pichia secretory yeast expression system. The recombinant protein was purified by heparin affinity chromatography. Law angiogenesis experiment and mouse wound healing test to detect activity. RESULTS: Recombinant human angiostatin was expressed in yeast at a high level (5 mg / L) and purified by heparin affinity chromatography. SDS PAGE showed a molecular weight of 43 kD. Activity experiments show that the recombinant protein can inhibit angiogenesis and inhibit wound healing. Conclusion: Recombinant human angiostatin is highly expressed in yeast and has good biological activity.