量子点的表面修饰、表征及其活细胞成像研究
首发时间:2016-05-16
摘要:目的 通过对疏水性量子点表面进行亲水性改性,并偶联单克隆抗体,制备免疫荧光探针用于细胞荧光标记示踪。方法 将BSA作为乳化剂分子对疏水性量子点进行表面亲水性改性。在检测亲水性改造后的量子点物理特性的同时,测定其对细胞活力的影响。随后将亲水性改造后的量子点与trastuzumab偶联,对HER2阳性乳腺癌细胞进行实时荧光成像检测。结果 经亲水性改造后的量子点粒径大约70 nm,分布较均一;在不同离子强度和pH环境中,保持良好的发光性能和胶体稳定性;生物相容性佳,未检测到明显细胞毒作用;经表面偶联肿瘤特异性单克隆抗体后,成功地对HER-2阳性乳腺癌细胞进行了长时间的活细胞跟踪成像研究。结论 亲水性量子点可通过选择性偶联抗体,获得免疫量子点荧光探针,从而对细胞、组织等进行特异性荧光成像和检测。
关键词: 量子点 牛血清白蛋白 表面改性 超声乳化 乳腺癌 免疫荧光
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Surface engineering and characterization of quantum dots and its application in living cell imaging
Abstract:Objective Surface modification of hydrophobic nanoparticles is a key process for their application in the biological medicine fields. The aim of this study is to investigate whether conjugation of surface-engineered QDs and monoclonal antibody could be used as immunofluorescent probes. Methods Considering excellent water-solubility and biocompatibility of bovine serum albumin (BSA), it was employed as the emulsifying agent in this study, and was used for the surface modification of hydrophobic quantum dots under ultrasonication. Diameter, fluorescence spectrum and the cytotoxicity of BSA-coated QDs (BSA-triP?QDs) were analyzed. Then BSA-coated QDs were conjugated with trastuzumab, which can recognize and bind specifically to HER2. SKBR-3 breast cancer cells, over-expression of HER-2, were labeled with hydrophilic QDs-trastuzumab and imaged on a fluorescence planar imaging system. Results The experimental results reveal that BSA as the emulsifying agent, combined with other polymer, can effectively phase transfer of hydrophobic quantum dots. The BSA functionalized QDs exhibit excellent colloidal stability with fine hydrodynamic size distribution (about 70 nm) in a wide range of pH and ionic strengths values. Moreover, no significant cytotoxic effect was observed in SKBR-3 cells treated with the BSA-coated QDs. After being coupled with trastuzumab, the hydrophilic quantum dots can be used as the immunofluorescence probe for HER2 postive breast cancer cell imaging. Conclusion The advantages of BSA-coated QDs include straight forward synthesis, high colloidal stability, and promising immunofluorescence characteristic when coupled with specific antibody. These are therefore proved to be ideal nano systems for biomedical labeling, targeting, and imaging.
Keywords: Quantum dots BSA Surface modification ultrasonic emulsification breast cancer immunofluorescence
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No.4687119114676714****
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