基于共轭高分子复合物能量转移的非标记DNA检测

doi:10.6023/A12100740

化学学报 ›› 2012, Vol. 70 ›› Issue (24): 2507-2512.DOI: 10.6023/A12100740 上一篇下一篇

研究论文

基于共轭高分子复合物能量转移的非标记DNA检测

邓洪平^a, 王国建^b, 朱邦尚^a, 朱利娟^b, 王大力^b, 庄园园^b, 朱新远^a,b

a 分析测试中心上海交通大学上海 200240;
b 化学化工学院上海交通大学上海 200240

投稿日期:2012-10-05 发布日期:2012-12-06
通讯作者: 朱新远 E-mail:xyzhu@sjtu.edu.cn; bshzhu@sjtu.edu.cn
基金资助:
项目受国家重点基础研究计划(Nos. 2012CB821500, 2013CB834506)、国家自然科学基金(No. 20974062)、上海市启明星计划(No. 11QH1401500)和国家杰出青年科学基金(No. 21025417)资助.

Label-Free DNA Detection through Energy Transfer of Conjugated Polymer Complexes

Deng Hongping^a, Wang Guojian^b, Zhu Bangshang^a, Zhu Lijuan^b, Wang Dali^b, Zhuang Yuanyuan^b, Zhu Xinyuan^a,b

a Instrumental Analysis Center, Shanghai Jiao Tong University, Shanghai 200240, China;
b School of Chemistry and Chemical Engineering, Shanghai Jiao Tong University, Shanghai 200240, China

Received:2012-10-05 Published:2012-12-06
Supported by:
Project supported by the National Basic Research Program (Nos. 2012CB821500, 2013CB834506), the National Natural Science Foundation of China (No. 20974062), Shanghai Rising-Star Program (No. 11QH1401500) and China National Funds for Distinguished Young Scientists (No. 21025417).

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摘要/Abstract

非标记DNA检测是一种高灵敏度、高选择性的DNA检测方法, 具有重要的科学和社会意义. 本文采用交叉偶联法制备了水溶性阳离子共轭聚合物: 聚(9,9-双(6'-N,N,N-三甲胺盐-己烷基)-芴亚苯基)(PFP); 利用氧化加成聚合反应制备了水溶性阴离子共轭聚合物: 聚(3-噻吩乙酸钠)(P3TSA). 通过核磁共振氢谱(¹H NMR)、傅立叶变换红外光谱(FTIR)等对其结构进行了表征. PFP与P3TSA通过静电相互作用形成稳定的高分子复合物. 利用紫外-可见光谱(UV-vis)和荧光发射光谱证明共轭高分子复合物能够发生能量转移. 保持PFP的浓度不变, 高分子复合物能量转移效率(ETEF)随着P3TSA浓度的增加而逐渐增大. 选取ETEF较高的样品, 考察了DNA探针用量对高分子复合物ETEF的影响. 随着DNA探针浓度的增加, ETEF逐渐减弱. 最后, 利用0.2 nmol DNA探针进行了DNA杂交配对检测. 实验结果表明, 这种检测方法可以明显区分完全互补配对、双碱基错配和非完全互补配对的目标DNA. 简而言之, 我们成功发展了一种基于共轭高分子复合物能量转移、具有高选择性的非标记DNA检测方法.

关键词: 共轭聚合物, DNA检测, 非标记, 高分子复合物, 能量转移

Label-free DNA detection is a highly sensitive and selective method with great scientific and social significance. Water-soluble cationic conjugated polymer (CCP), poly(9,9-bis(6'-N,N,N-trimethylammonium)-hexyl)-fluorene phenylene) (PFP), was synthesized by cross coupling reaction under the catalysis of palladium, and water-soluble anionic conjugated polymer (ACP), poly(3-thiophene sodium acetate) (P3TSA), was prepared through oxidative addition polymerization with anhydrous ferric chloride as the catalyst. Both PFP and P3TSA were well characterized by ¹H nuclear magnetic resonance (¹H NMR) and Fourier transform infrared (FTIR) tests. Owing to the existence of opposite ionic side-chains, PFP and P3TSA were both water-soluble and could form polymer complexes through electrostatic interactions. The transmittance measurements of the polymer complex solution with wavelength ranging from 600 to 750 nm were performed. The results showed that the transmittance was almost unchanged for all samples, confirming the high solubility and stability of polymer complexes. The ultraviolet-visible (UV-vis) and photoluminescence emission measurements showed that the emission spectrum of PFP overlapped a lot with the absorption spectrum of P3TSA, proving the energy transfer from PFP (donor) to P3TSA (acceptor). By fixing the donor concentration, energy transfer efficiency (ETEF) of the polymer complexes increased with the concentration of P3TSA. With the enhancement of P3TSA concentration, the ETEF increased slowly at first and then relatively fast at a high P3TSA concentration. A sample with a high ETEF was selected to investigate the effect of DNA probe (X₁) concentration on ETEF. It was observed that ETEF decreased with the increase of X₁ concentration, owing to the repulsion of P3TSA by negative charged DNA probes. DNA hybridization was conducted by using 0.2 nmol of X₁ as a probe. The experimental results showed that the perfectly matched, double mismatched and non-matched target DNA could be well distinguished with obvious photoluminescence signals. In conclusion, a new label-free DNA detection method with high selectivity has been successfully developed based on the energy transfer of conjugated polymer complexes.

Key words: conjugated polymer, DNA detection, label-free, polymer complexes, energy transfer

引用此文

邓洪平, 王国建, 朱邦尚, 朱利娟, 王大力, 庄园园, 朱新远. 基于共轭高分子复合物能量转移的非标记DNA检测[J]. 化学学报, 2012, 70(24): 2507-2512.

Deng Hongping, Wang Guojian, Zhu Bangshang, Zhu Lijuan, Wang Dali, Zhuang Yuanyuan, Zhu Xinyuan. Label-Free DNA Detection through Energy Transfer of Conjugated Polymer Complexes[J]. Acta Chimica Sinica, 2012, 70(24): 2507-2512.

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基于共轭高分子复合物能量转移的非标记DNA检测

Label-Free DNA Detection through Energy Transfer of Conjugated Polymer Complexes

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