磁性纳米粒子-抗体复合物分离肝癌线粒体

摘要

目的 利用人肝癌组织,研究磁性纳米粒子-抗体复合物与传统线粒体分离方法之间的差异.方法 取人肝癌组织,分别采用磁性纳米粒子-抗体复合物和密度梯度离心方法分离线粒体.扫描电镜观察线粒体形态,Western blot检测线粒体蛋白标志物,流式细胞仪分析线粒体膜电位变化.结果 采用磁性纳米粒子-抗体复合物方法分离得到的肝癌组织线粒体结构完整,无胞核、胞膜等组分的污染,几乎没有过氧化物酶体和溶酶体的污染.经流式细胞仪检测,线粒体膜电位稳定.结论 与传统方法相比,磁性纳米粒子-抗体复合物分离方法具备简便易行,分离得到的线粒体结构完整、纯度及活性较高,并且不需要大型设备等优点.%Objective To identify differences between magnetic nanoparticles-antibody complex and the traditional method to isolate mitochondria from human liver cancer tissues. Methods Using magnetic nanoparticles-antibody complex and density gradient centrifugation to isolate mitochondria of human liver cancer tissues separately, then scanning electron microscopy was used to scan the morphology of mitochondria, Western blot was used to detect mi-tochondrial protein markers and flow cytometry was used to analyze the alteration of mitochondrial membrane potential. Results The mitochondria isolated by magnetic nanoparticle-antibody had intergral structure, contained almost membrane no contaminant of nucleus, cell membrane, peroxisomes and lysosomes. The mitochondrial potential was stable. Conclusions Magnetic nanoparticle-antibody complex method gets more advantages, such as the preparation is easier, the structure of mitochondria is intact, the purity and activity of mitochondria are higher, expensive equipments are not needed.