毛细管电泳激光诱导荧光法优化量子点-转铁蛋白偶联体系及用于细胞标记  

Optimization of Quantum Dot System of CdTe and Transferrin by Capillary Electrophoresis-Laser Inductive Fluorescence and Verification in Cell Labeling

作　　者：梅芳[1] 赵新颖[1,2] 张璐[1] 屈锋[1] 

机构地区：[1]北京理工大学生命学院,北京100081 [2]北京市理化分析测试中心,北京100089

出　　处：《分析化学》2013年第5期725-731,共7页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(Nos.21175011;20875009);国家重点基础研究发展计划(No.2012CB0603)资助

摘　　要：利用毛细管电泳激光诱导荧光(CE-LIF)表征了偶联剂N-羟基硫代琥珀酰亚胺(NHS)及NHS和1-(3-二甲氨基丙基)-3-乙基碳二亚胺盐酸盐(EDC)混合偶联剂对CdTe量子点活化效果的差异,优化了CdTe与转铁蛋白(Trf)的偶联条件,比较了CdTe-Trf,CdTe-BSA偶联物及CdTe量子点对Hela细胞的标记效果。结果表明:NHS活化后的量子点性能优于混合偶联剂EDC-NHS。31.2μmol/L Trf与CdTe形成的偶联物CdTe-Trf对Hela细胞的标记效果最佳。CdTe-Trf在4℃孵育20 min可快速标记在Hela细胞的表面;在37℃孵育7 h后,可进入Hela细胞内。说明所制得的CdTe-Trf偶联物具有Trf的生物功能,它可通过特异性识别并结合细胞膜表面的Trf受体而转入Hela细胞。而CdTe-BSA偶联物的标记不具特异性,CdTe则不能用于Hela细胞的标记。Capillary electrophoresis-laser inductive fluorescence （CE-LIF） was used to characterize the active effect difference of coupling agent N-hydroxysulfo-succinimide （NHS） and the mixture of NHS and 1-ethyl-3-（3-dimethylaminopropyl） carbodiimides hydrochloride （EDC） on quantum dot CdTe, and to opti- mize the coupling conditions of CdTe and transferrin （Trf）. The labeling effect of CdTe-Trf and CdTe-BSA complex as well as free CdTe for Hela cell were compared. Results demonstrated that the property of CdTe activated by NHS was better than the mixture of EDC-NHS. When 12.5, 31.2 and 40.6 p.mol/L Trf were used to couple CdTe respectively, the resultant of CdTe with 31.2 p^mol/L Trf optimized by CE-LIF showed the best labeling on Hela. The CdTe-Trf resultant labeled Hela rapidly on surface when they incubated at 4 ~C for 20 min, and transferred into Hela when they incubated at 37℃ for 7 h, which manifested the CdTe-Trf resultant had biological function. The CdTe-Trf can recognize and bind the Trf receptor on cell membrane specifically and enter Hela, while CdTe-BSA can not recognize Hela specifically and free CdTe can not label on cell.

关 键 词：毛细管电泳-激光诱导荧光(CE-LIF) 量子点 转铁蛋白 偶联 细胞标记 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学] O657.3[医药卫生—基础医学]