半导体量子点-Smad2单克隆抗体探针检测大鼠牙乳头细胞内Smad2信号蛋白分子核移位过程

基础医学与临床 ›› 2009, Vol. 29 ›› Issue (8): 806-810.

半导体量子点-Smad2单克隆抗体探针检测大鼠牙乳头细胞内Smad2信号蛋白分子核移位过程

陈睿杨凯孙德平张国栋梅杰李雅冬

重庆医科大学附属第一医院重庆医科大学附属第一医院重庆医科大学附属第一医院

收稿日期:2008-08-28 修回日期:2008-11-03 出版日期:2009-08-20 发布日期:2009-08-20
通讯作者: 杨凯

The detection of the nucleation shifting process of Smad2 signal protein in Rat dental papillae cells with Quantum Dots-Smad2 monoclonal antibody Fluorescent probes

Rui CHEN, Kai YANG, De-ping SUN, Guo-dong ZHANG, Jie MEI, Ya-dong LI

the First Affiliated Hospital, Chongqing Medical University the First Affiliated Hospital, Chongqing Medical University

Received:2008-08-28 Revised:2008-11-03 Online:2009-08-20 Published:2009-08-20
Contact: Kai YANG,

摘要/Abstract

摘要： 目的制备半导体量子点-Smad2单克隆抗体荧光探针（QDS-Smad2），用其对大鼠牙乳头细胞内Smad2蛋白分子在TGF-β1刺激下发生的核移位过程进行检测。方法（1）用化学偶连法制备水溶性QDS-Smad2并纯化，检测其相关光学性质；（2）分别用QDS和Smad2单抗直接标记法和QDS-Smad2直接免疫荧光成像法观察QDS-Smad2对大鼠牙乳头细胞内Smad2的特异性识别能力，并检测细胞内QDS-Smad2的光学性质；（3）在大鼠牙乳头细胞内加入TGF-β1，分别于加入前、加入后12和24h，用QDS-Smad2直接免疫荧光成像法观察Smad2在细胞内发生核移位的动态变化。结果半导体量子点与Smad2单抗通过共价结合形成稳定的QDS-Smad2，QDS-Smad2对大鼠牙乳头细胞内Smad2分子仍具有特异性的免疫识别能力，能成像显示Smad2所产生核移位的动态变化；QDS-Smad2仍具有QDS所具有的荧光度强，光化学稳定性好的光学特征。结论半导体量子点和单抗共价结合形成分子探针后仍具有独特的光学性质和特异免疫识别能力，能长时间对细胞内蛋白质分子进行成像标记。

关键词: 量子点, 纳米技术, 牙乳头细胞, Smad2

Abstract: Objective To prepare Semiconductor Quantum Dots (QDs)-Smad2 monoclonal antibody Fluorescent probes , to detect nucleation shifting process of Smad2 signal protein in Rat dental papillae cells(RDPCs)stimulated by TGF-β1 with the Fluorescent probes.Methods ① QDs were chemically modified with Smad2 proteins to prepare water soluble QDs-Smad2 monoclonal antibody Fluorescent probes which were purified after the preparation, and detect its related optical properties. ②the location of Smad2 proteins in RDPCs was studied with QDs direct mark method、Smad2 monoclonal antibody direct mark method and direct immunofluorescence imaging. And the properties of Quantum Dots-Smad2 monoclonal antibody Fluorescent probes in RDPCs were detected. ③after RDPCs cultivated by TGF-β1 for 0 hours、12 hours、24 hours ,the dynamic nucleation shifting process of Smad2 signal protein in RDPCs was separately detected with direct immunofluorescence imaging.Result QDs and monoclonal antibody covalently bond to form the Fluorescent probes which could specifically and effectively recognize Smad2 proteins in RDPCs .It is to demonstrate the dynamic nucleation shifting process of Smad2 signal protein in RDPCs. These Fluorescent probes still had good high fluorescence intensity and photostability. conclusion QDs and monoclonal antibody could covalently bond to form the Fluorescent probes with distinct optics character、special immunity recognition capability and the ability of imaging mark the protein in cells for long time .

Key words: Quantum dots, Nanotechnology, Dental papillae cells, Smad2

陈睿杨凯孙德平张国栋梅杰李雅冬. 半导体量子点-Smad2单克隆抗体探针检测大鼠牙乳头细胞内Smad2信号蛋白分子核移位过程[J]. 基础医学与临床, 2009, 29(8): 806-810.

Rui CHEN; Kai YANG; De-ping SUN; Guo-dong ZHANG; Jie MEI; Ya-dong LI. The detection of the nucleation shifting process of Smad2 signal protein in Rat dental papillae cells with Quantum Dots-Smad2 monoclonal antibody Fluorescent probes[J]. Basic & Clinical Medicine, 2009, 29(8): 806-810.