化学学报 ›› 2013, Vol. 71 ›› Issue (06): 962-966.DOI: 10.6023/A12121072 上一篇    

研究论文

碳量子点荧光成像法应用于聚丙烯酰胺凝胶电泳检测人血清蛋白质的研究

刘亭廷a, 彭程a, 马云川a, 欧阳津b   

  1. a 首都医科大学宣武医院 北京 100053;
    b 北京师范大学化学学院 北京 100875
  • 收稿日期:2012-12-19 出版日期:2013-06-14 发布日期:2013-04-12
  • 通讯作者: 刘亭廷,lmt_52@sina.com; Tel.: 010-83198262 E-mail:lmt_52@sina.com

Application of Carbon Quantum Dots in Fluorescent Imaging of Human Serum Proteins after Polyacrylamide Gel Electrophoresis

Liu Tingtinga, Peng Chenga, Ma Yunchuana, Ouyang Jinb   

  1. a Xuanwu Hospital, Capital Medical University, Beijing 100053;
    b Beijing Normal University, Beijing 100875
  • Received:2012-12-19 Online:2013-06-14 Published:2013-04-12

建立了碳量子点荧光成像法检测聚丙烯酰胺凝胶电泳分离蛋白质如人血清蛋白质的新方法. 通过一步绿色微波法合成的碳量子点并将其应用在聚丙烯酰胺凝胶电泳分离蛋白质的检测中, 通过醋酸-醋酸钠调节孵育溶液, 优化pH值、碳量子点的用量及孵育溶液的浓度等条件, 使碳量子点和蛋白质相结合并在365 nm的紫外灯照射下得到了清晰的人血清蛋白电泳图, 该新方法具有原料便宜易得、无污染、简单、快速、高灵敏度、低背景及高分辨率的优点, 在生物技术方面和纳米技术方面具有巨大的应用前景.

关键词: 碳量子点, 聚丙烯酰胺凝胶电泳, 蛋白质, 荧光成像

A new method of carbon quantum dots (CQDs) fluorescent imaging for human serum proteins detection after polyacrylamide gels electrophoresis (PAGE) is established. Polyacrylamide gel electrophoresis is one of the most general and powerful technique to separate complex biosamples, and it is widely used in molecular biology, biochemistry and medicine. With the evolution of clinical proteomics, the development of a novel method to detect serum proteins after PAGE with high resolution and high sensitivity is of great significant. As far as we know, the carbon quantum dots have not been applied in the detection of serum proteins after PAGE. The fluorescent carbon quantum dots were synthesized by a one-step microwave pyrolysis method: glycerol and phosphate buffer (7.1 mmol•L-1, pH 7.4) (φ=70%) was mixed evenly, and then put the solution into the advanced microwave digestion system and heated for 14 min (750 W), the colorless solution turned to yellow after reaction. The yellow solution was diluted into the incubation solution with HOAc-NaOAc buffer (80 mmol•L-1, pH 2.7), and then used for staining serum proteins. The emission and excitation spectra of carbon quantum dots were measured, and the excitation wavelength allowed the use of an ultraviolet lamp at 365 nm for the fluorescent imaging. In order to investigate the performance of CQDs fluorescent imaging, the dilution of human serum samples separated by electrophoresis was used for detection, and was compared with traditional method of CBB-R250 staining and silver staining. The results suggested that CQDs based on fluorescent imaging could effectively detect human serum proteins with high sensitivity and resolution. The sensitivity of CQDs imaging was higher than CBB staining and comparable to silver staining. Therefore, the CQDs fluorescent imaging could be an inexpensive, time-saving, pollution-free and convenient method with high sensitivity and resolution for the detection of human serum proteins. It demonstrates the CQDs fluorescent imaging technology has great significant prospects in the development of biotechnology and nanotechnology.

Key words: carbon quantum dot, polyacrylamide gel electrophoresis, protein, fluorescent imaging