Anti-HER2免疫脂质体的制备及其靶向特异性的检测  

Prototype HER2 Immunoliposome and Characterization of Its Targeting Specificity

作　　者：崔潇 马亚平 张亚柯 李英 张芳霞 王瑞 CUI Xiao;MA Yaping;ZHANG Yake;LI Ying;ZHANG Fangxia;WANG Rui(School of Pharmacy,Liaocheng University,Liaocheng 252059,China)

机构地区：[1]聊城大学药学院,山东聊城252059

出　　处：《聊城大学学报（自然科学版）》2021年第2期96-102,共7页Journal of Liaocheng University:Natural Science Edition

基　　金：国家自然科学基金项目(31401217,81402512);大学生创新创业训练计划(201910447910)资助。

摘　　要：目的将靶向HER2过表达细胞的HER2抗体偶联在纳米脂质体上,探索抗体偶联脂质体的方法。方法通过乙醇注入法制备脂质体,表面修饰HER2抗体,成功制备Anti-HER2免疫脂质体;由SDS-PAGE电泳检测抗体偶联脂质体的连接效率;通过MTT检测免疫脂质体对乳腺癌细胞的细胞毒性;采用共聚焦显微镜和流式细胞技术检测免疫脂质体对乳腺癌细胞的靶向特异性。结果粒径分析表明制备的空白脂质体与Anti-HER2免疫脂质体粒径均在100-135 nm之间;SDS-PAGE电泳显示HER2抗体与脂质体的连接效率为40%左右;MTT检测表明制备的免疫脂质体具有低细胞毒性;共聚焦显微镜和流式细胞仪检测结果均显示,Anti-HER2免疫脂质体对HER2阳性细胞具有明显的靶向特异性。结论通过两种聚乙二醇连接方法成功制备了Anti-HER2免疫脂质体,数据表明DSPE-PEG2000-NHS(二硬脂酰磷脂酰乙酰胺-聚乙二醇2000-N-羟基丁二酰亚胺)连接方式效率更高。Objective:HER2 antibodies targeting HER2 overexpression cells were coupled to nano-liposomes to explore the method of antibody coupling liposomes.Methods:Liposomes were prepared by ethanol injection.HER2 antibody was modified on the surface of liposomes,and Anti-HER2 immunoliposomes were successfully prepared.The connection efficiency of antibody-coupled liposomes was detected by SDS-PAGE electrophoresis,the cytotoxicity of immunoliposomes to breast cancer cells was detected by MTT,and the targeting specificity of immune liposomes to to breast cancer cells was detected by confocal microscope and flow cytometry.Result:Particle size analysis showed that the average particle sizes of blank liposomes and Anti-HER2 immunoliposomes were between 100-135 nm.SDS-PAGE electrophoresis showed that the binding efficiency of HER2 antibody to liposome was about 40%.MTT detection showed that the prepared immunoliposomes had low cytotoxicity.Confocal microscopy and flow cytometry showed that Anti-HER2 modified immunoliposomes had obvious targeting specificity to HER2 positive cells.Conclusion:Anti-HER2 immunoliposomes were successfully prepared by two polyethylene glycol linking methods.The data showed that N-hydroxysuccinimide modified PEGylated liposomes was more efficient.

关 键 词：免疫脂质体 HER2抗体 特异性靶向 

分 类 号：R943[医药卫生—药剂学] R96[医药卫生—药学]