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目的:制备精子蛋白17(Sp17)抗体免疫磁性纳米颗粒,以期用于卵巢癌的磁共振成像靶向诊断。方法:在偶联剂EDC/NHS作用下,将壳聚糖修饰磁性氧化铁纳米颗粒与抗人Sp17抗体结合,制备抗Sp17抗体免疫磁性纳米颗粒(IMNPs)。透射电镜观察颗粒的形貌特征,非变性聚丙烯酰胺凝胶电泳评价抗体与磁性颗粒的结合效果,ELISA检测免疫磁性颗粒的免疫活性。将IMNPs与转染人Sp17的卵巢癌HO-8910细胞共孵育,进行体外磁共振成像检测。结果:成功实现了抗体与磁性纳米颗粒的偶联,并且IMNPs保持良好的抗体活性。磁共振显示该颗粒成功靶向Sp17阳性的细胞,没有发生明显的非特异吸附。结论:该免疫磁性纳米颗粒特异性良好,可进一步用作卵巢癌的靶向治疗研究。
OBJECTIVE: To prepare sperm protein 17 (Sp17) antibody immunomagnetic nanoparticles for the purpose of magnetic resonance imaging targeted diagnosis of ovarian cancer. Methods: The anti-Sp17 antibody immunomagnetic nanoparticles (IMNPs) were prepared by coupling chitosan-modified magnetic iron oxide nanoparticles with anti-human Sp17 antibody under the action of EDC / NHS coupling agent. Transmission electron microscopy (TEM) was used to observe the morphological features of the particles. Non-denaturing polyacrylamide gel electrophoresis was used to evaluate the binding of the antibody to the magnetic particles. ELISA was used to measure the immunological activity of the immunomagnetic particles. IMNPs were incubated with ovarian cancer HO-8910 cells transfected with human Sp17 for in vitro magnetic resonance imaging. Results: Coupling of antibodies to magnetic nanoparticles was successfully achieved and IMNPs retained good antibody activity. Magnetic resonance imaging showed that the particles successfully targeted Sp17 positive cells without significant nonspecific adsorption. Conclusion: The immunomagnetic nanoparticles have good specificity and can be further used as targeted therapy in ovarian cancer.