化学学报 ›› 2014, Vol. 72 ›› Issue (4): 440-446.DOI: 10.6023/A13121247 上一篇    下一篇

研究论文

生物素功能化水溶性共轭聚合物的合成、表征及其生物分析应用

刘兴奋a, 蔡小慧a, 黄艳琴a, 石琳a, 范曲立a, 黄维a,b   

  1. a 有机电子与信息显示国家重点实验室培育基地 南京邮电大学信息材料与纳米技术研究院 南京 210023;
    b 江苏-新加坡有机电子和信息显示联合实验室 南京工业大学先进材料研究院 南京 211816
  • 收稿日期:2013-12-20 出版日期:2014-04-14 发布日期:2014-02-14
  • 通讯作者: 刘兴奋 E-mail:iamxfliu@njupt.edu.cn; Tel.:025-85866396; Fax:025-85866396. E-mail:iamxfliu@njupt.edu.cn
  • 基金资助:
    项目受国家重点基础研究发展计划(Nos.2009CB930601,2012CB933301,2012CB723402)、国家自然科学基金(Nos.21005040,51073078,51173080)、“有机与生物光电子学”教育部创新团队(No.IRT1148)、江苏高校优势学科建设工程及江苏省普通高校研究生科研创新计划(No.CXLX12_0468)资助.

Biotinylated Water-Soluble Conjugated Polymers:Synthesis and the Application in Biological Analysis

Liu Xingfena, Cai Xiaohuia, Huang Yanqina, Shi Lina, Fan Qulia, Huang Weia,b   

  1. a Key Laboratory for Organic Electronics & Information Displays (KLOEID) and Institute of Advanced Materials (IAM), Nanjing University of Posts and Telecommunications, Nanjing 210023;
    b Jiangsu-Singapore Joint Research Center for Organic/Bio-Electronics & Information Displays and Institute of Advanced Materials, Nanjing Tech University, Nanjing 211816)
  • Received:2013-12-20 Online:2014-04-14 Published:2014-02-14
  • Supported by:
    Project supported by the National Basic Research Program of China (Nos. 2009CB930601, 2012CB933301, 2012CB723402), the National Natural Science Foundation of China (Nos. 21005040, 51073078, 51173080), the Ministry of Education of China (No. IRT1148), the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) and the Program of Scientific Innovation Research of College Graduate in Jiangsu Province (CXLX12_0468).

以四(三苯基膦)钯/碘化亚铜为催化剂,利用Sonogashira偶联反应,合成了一种主链含芴炔苯结构的侧链生物素(biotin)修饰的具有蓝光发射特性的水溶性共轭聚合物(PFPNBr-biotin). 其结构通过核磁共振和红外光谱进行表征;分子量(分布)利用凝胶渗透色谱进行表征. 研究了该聚合物在不同溶剂、不同浓度及不同pH环境中的光物理性质变化. 与未功能化的共轭聚合物相比,这种生物素功能化的聚合物对亲和素蛋白显示出较高的选择性. 目前,生物素-亲和素系统已在生物分析领域广泛使用,因此,该聚合物在生物分子的特异性检测、荧光标记、细胞及分子成像等领域具有较好的应用前景.

关键词: 生物素, 水溶性共轭聚合物, 生物功能化, 生物分析

Conjugated polymers with good photostability, high absorption coefficients and quantum yields have attracted great interest in biological analysis, molecular electronics and organic electronics. Biofunctional conjugated polymers have many advantages in bioanalysis, such as high selectivity and excellent tumor targeting capacity. Herein, a biotinylated water-soluble conjugated polyfluorene derivative (PFPNBr-biotin) with blue fluorescence emission characteristic was synthesized via Sonogashira coupling reaction by using Pd(PPh3)4/CuI as catalysts. The molecular structure of compounds, neutral polymer and cationic polymer were characterized by nuclear magnetic resonance (NMR) and Fourier transform infrared spectroscopy (FT-IR); molecule weight and distribution of polymers were characterized by gel permeation chromatography (GPC) and other methods. The photophysical properties of the PFPNBr-biotin in mixed solvent systems with different ratio of methanol to water were studied by monitoring its fluorescence spectra. The results demonstrate an increased fluorescence intensity of PFPNBr-biotin with the increase of methanol. Changes in the photophysical properties of PFPNBr-biotin with the increasing polymer concentration were investigated by measuring UV-vis absorbance and fluorescence spectra. Some aggregates may form when the concentration of polymer is more than 400 μmol·L-1, which corresponding to decreased UV-vis absorbance and fluorescence emission. Optical properties of PFPNBr-biotin itself and the protein detection with PFPNBr-biotin in solutions of different pHs were studied to analyze the aggregation behavior before and after interacting with protein. Compared to the nonfunctional conjugated polymer, PFPNBr-biotin shows excellent selectivity to avidin over negative proteins such as lysozyme (Lys), immunoglobin G (IgG), hemoglobin (Hb) and bovine serum albumin (BSA). So it can be used to distinguish avidin (or strapatavidin) over many proteins just by monitoring the fluorescence change of PFPNBr-biotin. Considering the fact that biotin-avidin system has been widely used in biological analysis, we believe that this biotinylated water-soluble conjugated polymer has a potential application in detecting and labeling of biomolecules and fluorescence imaging of tumor cells.

Key words: biotin, water-soluble conjugated polymers, biofunctionalization, bioanalysis