Citation: Liu Xingfen, Wang Yateng, Huang Yanqin, Feng Xiaomiao, Fan Quli, Huang Wei. Highly Sensitive Protein Biosensor based on a Conjugated Polymer Brush[J]. Acta Chimica Sinica, ;2016, 74(8): 664-668. doi: 10.6023/A16040205 shu

Highly Sensitive Protein Biosensor based on a Conjugated Polymer Brush

Liu Xingfen ^a,*,, ,
Wang Yateng ^a ,
Huang Yanqin ^a ,
Feng Xiaomiao ^a ,
Fan Quli ^a ,
Huang Wei ^a,b

a.
Key Laboratory for Organic Electronics & Information Displays and Institute of Advanced Materials, Jiangsu National Synergetic Innovation Center for Advanced Materials, Nanjing University of Posts and Telecommunications, Nanjing 210023, China
b.
Key Laboratory of Flexible Electronics & Institute of Advanced Materials, Jiangsu National Synergetic InnovationCenter for Advanced Materials, Nanjing Tech University Nanjing Tech, Nanjing 211816, China

Corresponding author: Liu Xingfen, iamxfliu@njupt.edu.cn
Received Date: 25 April 2016

Fund Project: the Natural Science Foundation of Jiangsu Province BK20141424Research Program of Nanjing University of Posts and Telecommunications NY215171Program of Scientific Innovation Research of College Graduate in Jiangsu Province CXLX12_0792the National Natural Science Foundation of China 51173080the Ministry of Education of China IRT1148the National Natural Science Foundation of China 21005040the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD) BK20141424the National Basic Research Program of China 2012CB933301the National Basic Research Program of China 2012CB723402

Figures(5)

Simple and sensitive detection of proteins is crucial in biological analysis and medical diagnosis. Conjugated polymers (CPs) with π-conjugated backbones were recognized as having excellent light-harvesting capability and high fluorescent quantum yield. They have been widely used as an energy donor to amplify fluorescence signal via high efficient Föster resonance energy transfer (FRET). In particular, conjugated polymer brush with high charge density provides more possibilities due to stronger electrostatic interactions with negatively charged biomolecules. Here, we developed a highly sensitive protein biosensor for thrombin detection based on a conjugated polymer brush (PFNI) and a fluorescein-labeled aptamer (FAM-apt15). PFNI is a water-soluble cationic polyfluorene derivate with extremely high charge density (78 positive charges per repeat unit). PFNI can attract negatively charged aptamer through strong electrostatic interactions. In this case, the energy donor (PFNI) and acceptor (FAM) are in a close proximity, which results in an efficient FRET process and a high FRET signal. However, when the FAM-apt15 combines with the target protein, a rigid and big-sized G-quadruplex/thrombin complex formed. Due to the steric hindrance from the densely brush of PFNI, the distance between the two fluorophores increased significantly, leading to an inefficient FRET process and a low FRET signal. The strategy exhibits excellent specificity and the limit of detection (LOD) for thrombin in buffer was estimated to be 0.05 nmol/L. It also works well in diluted serum and a LOD of 0.2 nmol/L can be obtained. Compared to the biosensors based on traditional linear conjugated polymers, the sensitivity was improved by one order of magnitude. In addition, our strategy also shows the merits of simple, label-free, and low-cost because labeled DNA is much more expensive than unlabeled one. Based on the specific binding of aptamer and protein, this novel method can be extended to a highly sensitive detection of more proteins.
- conjugated polymer,
- aptamer,
- Föster resonance energy transfer,
- thrombin,
- biosensor
1. [1]
  Cera, E. D.; Dang, Q. D.; Ayala, Y. M. Cell Mol. Life Sci. 1997, 53, 701. doi: 10.1007/s000180050091
2. [2]
  Maragoudakis, M. E.; Tsopanoglou, N. E.; Andriopoulou, P. Biochem. Soc. T. 2002, 30, 173. doi: 10.1042/bst0300173
3. [3]
  Liu, C. W.; Huang, C. C.; Chang, H. T. Anal. Chem. 2009, 81, 2383. doi: 10.1021/ac8022185
4. [4]
  Song, W.; Zhang, Q.; Xie, X.; Zhang, S. Biosens. Bioelectron. 2014, 61, 51. doi: 10.1016/j.bios.2014.04.030
5. [5]
  Pavlov, V.; Xiao, Y.; Shlyahovsky, B.; Willner, I. J. Am. Chem. Soc. 2004, 126, 11768. doi: 10.1021/ja046970u
6. [6]
  Liang, H. R.; Hu, G. Q.; Xue, X. H.; Li, L.; Zheng, X. X.; Gao, Y. W.; Yang, S. T.; Xia, X. Z. Virus Res. 2014, 184, 7. doi: 10.1016/j.virusres.2014.01.021
7. [7]
  Kim, Y. S.; Song, M. Y.; Jurng, J.; Kim, B. C. Anal. Biochem. 2013, 436, 22. doi: 10.1016/j.ab.2013.01.014
8. [8]
  Cao, H. Y.; Yuan, A. H.; Shi, X. S.; Chen, W.; Miao, Y. Oncol. Rep. 2014, 32, 2054.
9. [9]
  Song, Q. W.; Peng, M. S.; Wang, L.; He, D. C.; Ouyang, J. Biosens. Bioelectron. 2016, 77, 237. doi: 10.1016/j.bios.2015.09.008
10. [10]
  Neves Miguel, A. D.; Blaszykowski, C.; Thompson, M. Anal. Chem. 2016, 88, 3098. doi: 10.1021/acs.analchem.5b04010
11. [11]
  Jiang, L. Y.; Xiao, X. N.; Zhou, P. L.; Zhang, P.; Yan, Y. X.; Jiang, S. X.; Chen, Q. H. Chinese J. Anal. Chem. 2016, 44, 310.
12. [12]
  Ge, J.; Liu, Z. F.; Zhao, X. S. Chinese J. Chem. 2012, 30, 2023. doi: 10.1002/cjoc.201200256
13. [13]
  Zhang, S. B.; Zheng, L. Y.; Hu, X.; Shen, G. Y.; Liu, X. W.; Shen, G. L.; Yu, R. Q. Chinese J. Anal. Chem. 2015, 43, 1688. doi: 10.1016/S1872-2040(15)60880-5
14. [14]
  Wu, C.; Yang, S. Y.; Wu, Z. Y.; Shen, G. L.; Yu, R. Q. Acta Chim. Sinica 2013, 71, 367. doi: 10.6023/A12110962
15. [15]
  He, F.; Tang, Y. L.; Wang, S.; Li, Y. L.; Zhu, D. B. J. Am. Chem. Soc. 2005, 127, 12343. doi: 10.1021/ja051507i
16. [16]
  Chen, Z. B.; Tan, L. L.; Hu, L. Y.; Zhang, Y. M.; Wang, S. X.; Lv, F. Y. ACS Appl. Mater. Inter. 2016, 8, 102. doi: 10.1021/acsami.5b08975
17. [17]
  Liu, X. F.; Shi, L.; Hua, X. X.; Fan, Q. L.; Chao, J.; Su, S.; Huang, Y. Q.; Wang, L. H.; Huang, W. ACS Appl. Mater. Inter. 2015, 7, 16458. doi: 10.1021/acsami.5b03662
18. [18]
  Kong, L.; Xu, J.; Xu, Y.; Xiang, Y.; Yuan, R.; Chai, Y. Biosens. Bioelectron. 2013, 42, 193. doi: 10.1016/j.bios.2012.10.064
19. [19]
  Chen, Z. B.; Tan, Y.; Zhang, C. M.; Yin, L.; Ma, H.; Ye, N. S.; Qiang, H.; Lin, Y. Q. Biosens. Bioelectron. 2014, 56, 46 doi: 10.1016/j.bios.2014.01.012
20. [20]
  Baek, S. H.; Wark, A. W.; Lee, H. J. Anal. Chem. 2014, 86, 9824. doi: 10.1021/ac5024183
21. [21]
  Jiang, C. N.; Liang, A. H.; Jiang, Z. L. Acta Chim.Sinica 2011, 69, 713.
22. [22]
  Le Floch, F.; Ho, H. A.; Leclerc, M. Anal. Chem. 2006, 78, 4727. doi: 10.1021/ac0521955
23. [23]
  Zhao, J.; Hu, S. S.; Zhong, W. D.; Wu, J. G.; Shen, Z. M.; Chen, Z.; Li. G. X. ACS Appl. Mater. Inter. 2014, 6, 7070. doi: 10.1021/am502053d
24. [24]
  Liu, L. Z.; Liu, Z. H.; He, Z. K.; Cai, R. X. Prog. Chem. 2006, 18, 337.
25. [25]
  Huang, F.; Wu, H. B.; Cao, Y. Chem. Soc. Rev. 2010, 39, 2500. doi: 10.1039/b907991m
26. [26]
  Wang, M.; Li, C. H.; Lv, A. F.; Wang, Z. H.; Bo, Z. S. Macromolecules 2012, 45, 3017. doi: 10.1021/ma202752h
27. [27]
  Thomas, S. W.; Joly, G. D.; Swagger, T. M. Chem. Rev. 2007, 107, 1339. doi: 10.1021/cr0501339
28. [28]
  Rochat, S.; Swagger, T. M. ACS Appl. Mater. Inter. 2013, 5, 4488. doi: 10.1021/am400939w
29. [29]
  Zhu, C. L.; Liu, L. B.; Yang, Q.; Lv, F. T.; Wang, S. Chem. Rev. 2012, 112, 4687. doi: 10.1021/cr200263w
30. [30]
  Wang, M. F.; Zou, S.; Guerin, G. Macromolecules 2008, 41, 6993 doi: 10.1021/ma800777m
31. [31]
  Pu, K. Y.; Li, K.; Liu, B. Adv. Funct. Mater. 2010, 20, 2770. doi: 10.1002/adfm.201000495
32. [32]
  Zhang, Z. Y.; Lu, X. M.; Fan, Q. L.; Hu, W. B.; Huang, W. Polym. Chem. 2011, 2, 2369. doi: 10.1039/c1py00213a
33. [33]
  Liu, X. F.; Shi, L.; Zhang, Z. Y.; Fan, Q. L.; Huang, Y. Q.; Su, S.; Fan, C. H.; Wang, L. H.; Huang, W. Analyst 2015, 140, 1842. doi: 10.1039/C4AN02384F
34. [34]
  Hu, W. B.; Lu, X. M.; Jiang, R. C.; Fan, Q. L.; Zhao, H.; Deng, W. X.; Zhang, L.; Huang, L.; Huang, W. Chem. Commun. 2013, 49, 9012. doi: 10.1039/c3cc45400b
35. [35]
  Jiang, R. C.; Lu, X. M.; Yang, M. H.; Deng, W. X.; Fan, Q. L.; Huang, W. Biomacromolecules 2013, 14, 3643. doi: 10.1021/bm401000x
36. [36]
  Zhang, Z. Y. Ph.D. Dissertation, Nanjing University of Posts and Telecommunications, Nanjing, 2014.
37. [37]
  Zhang, L. B.; Zhu, J. B.; Li, T.; Wang, E. K. Anal. Chem. 2011, 83, 8871. doi: 10.1021/ac2006763
38. [38]
  Golub, E.; Freeman, R.; Willner, I. Anal. Chem. 2013, 85, 12126. doi: 10.1021/ac403305k
39. [39]
  Lin, Z. H.; Pan, D.; Hu, T. Y.; Liu, Z. P.; Su, X. G. Microchim. Acta 2015, 182, 1933. doi: 10.1007/s00604-015-1526-4
40. [40]
  Yang, X. H.; Wang, S. F.; Wang, K. M.; Luo, X. M.; Tan, W. H.; Cui, L. Chem. J. Chinese Univ. 2009, 30, 899.
41. [41]
  Wang, Y. Y.; Liu, B. Langmuir 2009, 25, 12787. doi: 10.1021/la901703p
42. [42]
  Liu, X. F.; Shi, L.; Hua, X. X.; Huang, Y. Q.; Su, S.; Fan, Q. L.; Wang, L. H.; Huang, W. ACS Appl. Mater. Inter. 2014, 6, 3406. doi: 10.1021/am405550j
1. [1]
  Qinqi Zhang , Zhuoran Bi , Yi Zhong , Ziting Xu , Lanlan Chen . 双重信号放大功能核酸荧光探针用于水中抗生素的超灵敏检测. University Chemistry, 2026, 41(5): 448-454. doi: 10.12461/PKU.DXHX202511151
2. [2]
  Yang Meiqing , Lu Wang , Haozi Lu , Yaocheng Yang , Song Liu . Recent Advances of Functional Nanomaterials for Screen-Printed Photoelectrochemical Biosensors. Acta Physico-Chimica Sinica, 2025, 41(2): 100018-0. doi: 10.3866/PKU.WHXB202310046
3. [3]
  Junjie Zhang , Chaochao Dong , Jinke Li , Jiawen Guo , Yingxin Shui , Yue Wang , Xinhua Duan . 共轭聚合物探针的设计及其对藻蓝蛋白的智能检测. University Chemistry, 2026, 41(5): 285-293. doi: 10.12461/PKU.DXHX202509114
4. [4]
  Mian Wei , Chang Cheng , Bowen He , Bei Cheng , Kezhen Qi , Chuanbiao Bie . Inorganic-organic CdS/YBTPy S-scheme photocatalyst for efficient hydrogen production and its mechanism. Acta Physico-Chimica Sinica, 2025, 41(12): 100158-0. doi: 10.1016/j.actphy.2025.100158
5. [5]
  Zhongxin YU , Wei SONG , Yang LIU , Yuxue DING , Fanhao MENG , Shuju WANG , Lixin YOU . Fluorescence sensing on chlortetracycline of a Zn-coordination polymer based on mixed ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2415-2421. doi: 10.11862/CJIC.20240304
6. [6]
  Xingchao Zhao , Xiaoming Li , Ming Liu , Zijin Zhao , Kaixuan Yang , Pengtian Liu , Haolan Zhang , Jintai Li , Xiaoling Ma , Qi Yao , Yanming Sun , Fujun Zhang . Photomultiplication-Type All-Polymer Photodetectors and Their Applications in Photoplethysmography Sensor. Acta Physico-Chimica Sinica, 2025, 41(1): 100007-0. doi: 10.3866/PKU.WHXB202311021
7. [7]
  You Wu , Chang Cheng , Kezhen Qi , Bei Cheng , Jianjun Zhang , Jiaguo Yu , Liuyang Zhang . Efficient Photocatalytic Production of H₂O₂ over ZnO/D-A Conjugated Polymer S-scheme Heterojunction and Charge Transfer Dynamics Investigation. Acta Physico-Chimica Sinica, 2024, 40(11): 2406027-0. doi: 10.3866/PKU.WHXB202406027
8. [8]
  Xiao SANG , Qi LIU , Jianping LANG . Synthesis, structure, and fluorescence properties of Zn(Ⅱ) coordination polymers containing tetra-alkenylpyridine ligands. Chinese Journal of Inorganic Chemistry, 2024, 40(11): 2124-2132. doi: 10.11862/CJIC.20240158
9. [9]
  Minghui Wu , Markus Mühlinghaus , Xuechao Li , Chaojie Xu , Qiang Chen , Haiming Zhang , Klaus Müllen , Lifeng Chi . On-Surface Synthesis of Chevron-Shaped Conjugated Ladder Polymers Consisting of Benzo[a]azulene Units. Acta Physico-Chimica Sinica, 2024, 40(8): 2307024-0. doi: 10.3866/PKU.WHXB202307024
10. [10]
  Hanmei Lü , Xin Chen , Qifu Sun , Ning Zhao , Xiangxin Guo . Uniform Garnet Nanoparticle Dispersion in Composite Polymer Electrolytes. Acta Physico-Chimica Sinica, 2024, 40(3): 2305016-0. doi: 10.3866/PKU.WHXB202305016
11. [11]
  Botao QU , Qian WANG , Qian WANG , Ruiping ZHANG . Synthesis, crystal structures, and luminescence properties of zinc coordination polymers based on 2,5-dibromoterephthalate. Chinese Journal of Inorganic Chemistry, 2026, 42(5): 917-924. doi: 10.11862/CJIC.20260357
12. [12]
  Qiaoqiao BAI , Anqi ZHOU , Xiaowei LI , Tang LIU , Song LIU . Construction of pressure-temperature dual-functional flexible sensors and applications in biomedicine. Chinese Journal of Inorganic Chemistry, 2024, 40(12): 2259-2274. doi: 10.11862/CJIC.20240128
13. [13]
  Fanpeng Meng , Fei Zhao , Jingkai Lin , Jinsheng Zhao , Huayang Zhang , Shaobin Wang . Optimizing interfacial electric fields in carbon nitride nanosheet/spherical conjugated polymer S-scheme heterojunction for hydrogen evolution. Acta Physico-Chimica Sinica, 2025, 41(8): 100095-0. doi: 10.1016/j.actphy.2025.100095
14. [14]
  Yuxia Luo , Xiaoyu Xie , Fangfang Chen . 药物递送魔法师——分子印迹聚合物. University Chemistry, 2025, 40(8): 202-210. doi: 10.12461/PKU.DXHX202409129
15. [15]
  Ke Zhao , Zhen Liu , Luyao Liu , Changyuan Yu , Jingshun Pan , Xuguang Huang . Functionalized Reflective Structure Fiber-Optic Interferometric Sensor for Trace Detection of Lead Ions. Acta Physico-Chimica Sinica, 2024, 40(4): 2304029-0. doi: 10.3866/PKU.WHXB202304029
16. [16]
  Kaimin WANG , Na HE , Shiyi LI , Xuling BAI , Weiqing SUN , Yanqing YE , Yulu MA . Synthesis, Hirshfeld surface analysis and properties of two Zn(Ⅱ)/Ni(Ⅱ) coordination polymers. Chinese Journal of Inorganic Chemistry, 2026, 42(1): 55-64. doi: 10.11862/CJIC.20250178
17. [17]
  Zhao Gao , Jianxiang Sun , Bin Mu , Yi Yan , Wei Tian . Exploration of Supramolecular Polymer “Chameleons” in Innovative Chemistry Experiments. University Chemistry, 2026, 41(4): 349-355. doi: 10.12461/PKU.DXHX202504044
18. [18]
  Cong Liu , Yuling Huang , Peng Zhen , Hualiu Li , Bingjia Xu , Suilian Luo , Guang Shi . A Comprehensive Design Experiment: Preparation and Property Characterization of Polymer Room-Temperature Phosphorescence Materials. University Chemistry, 2026, 41(4): 409-416. doi: 10.12461/PKU.DXHX202503092
19. [19]
  Xinyi MA , Yuhang XIAO , Qian LI , Zihuan YAN , Chengyan LIU , Xiguang GAO , Hongju YIN , Feixiang CHENG . Thienyl metal coordination polymers: Preparation and photocatalytic degradation performance of phenol. Chinese Journal of Inorganic Chemistry, 2026, 42(5): 969-979. doi: 10.11862/CJIC.20250317
20. [20]
  Lin′an CAO , Dengyue MA , Gang XU . Research advances in electrically conductive metal-organic frameworks-based electrochemical sensors. Chinese Journal of Inorganic Chemistry, 2025, 41(10): 1953-1972. doi: 10.11862/CJIC.20250160

Get Citation

PDF

XML

Metrics

PDF Downloads(0)
Abstract views(1546)
HTML views(239)

通讯作者: 陈斌, bchen63@163.com

1.
沈阳化工大学材料科学与工程学院沈阳 110142