精子蛋白17抗体免疫磁性颗粒活性分析及细胞成像研究  

Biological activity assays and cellular imaging of anti-human sperm protein 17 immunomagnetic nanoparticles

作　　者：刘群[1] 葛玉卿[2] 李芳秋[1] 张士新[1] 顾宁[2] 王中秋[3] 卢光明[3] 

机构地区：[1]南京军区南京总医院解放军临床检验医学研究所,江苏南京210002 [2]东南大学生物科学与医学工程学院,江苏南京210096 [3]南京军区南京总医院医学影像科,江苏南京210002

出　　处：《细胞与分子免疫学杂志》2009年第11期987-990,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30670599);国家重点基础研究发展规划(2006CB705707);南京军区医学科学技术"十一五"研究计划重点项目(06Z43);国家自然科学基金杰出青年基金项目(60725101);国家重大科学研究计划项目(2006CB933206)

摘　　要：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。AIM： To prepare anti-Sperm protein 17 （ Sp17 ） immunomagnetic nanoparticles （ IMNPs）, and make foundation for target diagnosis of ovarian cancer by magnetic resonance imaging. METHODS： The anti-human Sp17 IMNPs were prepared by grafting anti-Sp17 antibodies on the surface of chitosan-coated magnetic nanoparticles （MNPs） using the linker of EDC/NHS （l-ethyl-3-[ 3-dimethylaminopropyl] carbodiimide/N-hydroxysuccinimide ）. The morphology and properties of the nanoparticles were characterized by transmission electronic microscopy （TEM）, the conjugation of the antibodies was evaluated by native-polyacrylamide gel electrophoresis, the immunologic activity of IMNPs was evaluated by enzyme linked immunosorbent assay （ELISA）. A set of in vitro magnetic resonance imaging （MRI） experiments were performed after incubated the IM- NPs with human Sp17 gene transfected ovarian cancer HO- 8910 cells. RESULTS： We had successfully grafted the MNPs with anti-Sp17 antibody and the IMNPs kept good bioactivity. The MRI showed that the IMNPs were targeted successfully to the positive cells, and no obviously non-specific adsorption was observed. CONCLUSION： The anti-SpiT IMNPs with good specificity can used for further study of ovarian cancer target therapy.

关 键 词：精子蛋白17抗体 免疫磁性颗粒 活性分析 磁共振成像 

分 类 号：R392.11[医药卫生—免疫学]