热沉淀纯化对促凋亡铁蛋白纳米粒活性的影响

Effects of thermal precipitation purification on the activity of pro-apoptotic ferritin nanoparticles

目的设计促凋亡铁蛋白纳米粒BAK-HFn,并探究热沉淀纯化对其活性的影响。方法设计与构建促凋亡活性蛋白纳米粒BAK-HFn,验证其成功可溶表达,采用热沉淀纯化以及非热沉淀纯化两种方式对其进行纯化,并用动态光散射法测定纯化后纳米粒的粒径,采用CCK-8法检测纯化后BAK-HFn对肿瘤细胞C6的细胞毒性。结果用热沉淀纯化后的BAK-HFn蛋白几乎不抑制C6细胞的生长,而非热沉淀纯化法获得的BAK-HFn蛋白促凋亡的活性明显提高。结论热沉淀纯化会使经过BAK修饰的促凋亡铁蛋白纳米粒降低甚至丧失活性。

OBJECTIVE To design pro-apoptotic ferritin nanoparticles BAK-HFn,to investigate effects of thermal precipitation purification on their activities.METHODS Pro-apoptotic active protein nanoparticles BAK-HFn were designed and constructed,and their successful soluble expression was verified,followed purification by both thermal precipitation and non-thermal precipitation,as well as measurement of particle size using dynamic light scattering method.Additionally,cytotoxicity of the purified BAK-HFn on tumor cells C6 was detected through CCK-8 assay.RESULTS BAK-HFn protein purified through thermal precipitation showed almost no growth inhibitory effect on C6 cells,and the pro-apoptotic activity of BAK-HFn protein obtained using non-thermal precipitation purification method was significantly increased.CONCLUSION Thermal precipitation purification decreases or even eliminates the activity of BAK-modified pro-apoptotic ferritin nanoparticles.