基于水溶性共轭聚合物分子刷的高灵敏凝血酶生物传感器

doi:10.6023/A16040205

化学学报 ›› 2016, Vol. 74 ›› Issue (8): 664-668.DOI: 10.6023/A16040205 上一篇下一篇

研究论文

基于水溶性共轭聚合物分子刷的高灵敏凝血酶生物传感器

刘兴奋^a, 王亚腾^a, 黄艳琴^a, 冯晓苗^a, 范曲立^a, 黄维^a,b

a 南京邮电大学信息材料与纳米技术研究院有机电子与信息显示国家重点实验室培育基地江苏省有机电子和信息显示协同创新中心南京 210023;
b 南京工业大学先进材料研究院江苏省柔性电子重点实验室先进生物与化学制造协同创新中心南京 211816

投稿日期:2016-04-25 发布日期:2016-08-10
通讯作者: 刘兴奋 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项目（Nos. 2012CB933301，2012CB723402）、国家自然科学基金（Nos. 21005040、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资助项目、江苏省自然科学基金（BK20141424）、江苏省普通高校研究生科研创新计划项目（CXLX12_0792）及南京邮电大学校科研项目（NY215171）资助.

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

Received:2016-04-25 Published:2016-08-10
Supported by:
Project supported by the National Basic Research Program of China (Nos. 2012CB933301, 2012CB723402), the National Natural Science Foundation of China (Nos. 21005040, 51173080), the Ministry of Education of China (No. IRT1148), the Priority Academic Program Development of Jiangsu Higher Education Institutions (PAPD), the Natural Science Foundation of Jiangsu Province (BK20141424), Program of Scientific Innovation Research of College Graduate in Jiangsu Province (CXLX12_0792) and Research Program of Nanjing University of Posts and Telecommunications (NY215171).

文章分享

摘要/Abstract

以刷子状水溶性共轭聚芴（PFNI）为传感材料，以荧光素标记的核酸适体（FAM-apt15）为探针，设计了一种检测凝血酶的高灵敏度蛋白质传感器. PFNI的刷状结构带有大量正电荷，与负电荷的柔性单链核酸探针形成静电复合物，使能量供体（PFNI）与受体（FAM）之间的距离较近，发生高效荧光共振能量转移（Föster resonance energy transfer，FRET）. 当探针与靶凝血酶结合时，形成刚性且体积较大的G-四链体/凝血酶复合物，由于体积位阻和密集的刷子的阻碍作用，PFNI与FAM之间的距离被拉大，FRET效率显著降低. 对缓冲溶液中凝血酶检测的最低检测限可达0.05 nmol/L. 与基于线型共轭聚合物的蛋白质检测方法相比，灵敏度提高了至少一个数量级.

关键词: 共轭聚合物, 核酸适体, 荧光共振能量转移, 凝血酶, 生物传感器

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.

Key words: conjugated polymer, aptamer, Föster resonance energy transfer, thrombin, biosensor

引用此文

刘兴奋, 王亚腾, 黄艳琴, 冯晓苗, 范曲立, 黄维. 基于水溶性共轭聚合物分子刷的高灵敏凝血酶生物传感器[J]. 化学学报, 2016, 74(8): 664-668.

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

导出引用 EndNote|Reference Manager|ProCite|BibTeX|RefWorks

分享此文

参考文献

[1] Cera, E. D.; Dang, Q. D.; Ayala, Y. M. Cell Mol. Life Sci. 1997, 53, 701.
[2] Maragoudakis, M. E.; Tsopanoglou, N. E.; Andriopoulou, P. Biochem. Soc. T. 2002, 30, 173.
[3] Liu, C. W.; Huang, C. C.; Chang, H. T. Anal. Chem. 2009, 81, 2383.
[4] Song, W.; Zhang, Q.; Xie, X.; Zhang, S. Biosens. Bioelectron. 2014, 61, 51.
[5] Pavlov, V.; Xiao, Y.; Shlyahovsky, B.; Willner, I. J. Am. Chem. Soc. 2004, 126, 11768.
[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.
[7] Kim, Y. S.; Song, M. Y.; Jurng, J.; Kim, B. C. Anal. Biochem. 2013, 436, 22.
[8] Cao, H. Y.; Yuan, A. H.; Shi, X. S.; Chen, W.; Miao, Y. Oncol. Rep. 2014, 32, 2054.
[9] Song, Q. W.; Peng, M. S.; Wang, L.; He, D. C.; Ouyang, J. Biosens. Bioelectron. 2016, 77, 237.
[10] Neves Miguel, A. D.; Blaszykowski, C.; Thompson, M. Anal. Chem. 2016, 88, 3098.
[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. (姜利英, 肖小楠, 周鹏磊, 张培, 闫艳霞, 姜素霞, 陈青华, 分析化学, 2016, 44, 310.)
[12] Ge, J.; Liu, Z. F.; Zhao, X. S. Chinese J. Chem. 2012, 30, 2023.
[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. (张松柏, 郑丽英, 胡霞, 沈广宇, 刘学文, 沈国励, 俞汝勤, 分析化学, 2015, 43, 1688.)
[14] Wu, C.; Yang, S. Y.; Wu, Z. Y.; Shen, G. L.; Yu, R. Q. Acta Chim. Sinica 2013, 71, 367. (吴超, 杨胜园, 吴朝阳, 沈国励, 俞汝勤, 化学学报, 2013, 71, 367.)
[15] He, F.; Tang, Y. L.; Wang, S.; Li, Y. L.; Zhu, D. B. J. Am. Chem. Soc. 2005, 127, 12343.
[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.
[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.
[18] Kong, L.; Xu, J.; Xu, Y.; Xiang, Y.; Yuan, R.; Chai, Y. Biosens. Bioelectron. 2013, 42, 193.
[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
[20] Baek, S. H.; Wark, A. W.; Lee, H. J. Anal. Chem. 2014, 86, 9824.
[21] Jiang, C. N.; Liang, A. H.; Jiang, Z. L. Acta Chim. Sinica 2011, 69, 713. (蒋彩娜, 梁爱惠, 蒋治良, 化学学报, 2011, 69, 713.)
[22] Le Floch, F.; Ho, H. A.; Leclerc, M. Anal. Chem. 2006, 78, 4727.
[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.
[24] Liu, L. Z.; Liu, Z. H.; He, Z. K.; Cai, R. X. Prog. Chem. 2006, 18, 337.
[25] Huang, F.; Wu, H. B.; Cao, Y. Chem. Soc. Rev. 2010, 39, 2500.
[26] Wang, M.; Li, C. H.; Lv, A. F.; Wang, Z. H.; Bo, Z. S. Macromolecules 2012, 45, 3017.
[27] Thomas, S. W.; Joly, G. D.; Swagger, T. M. Chem. Rev. 2007, 107, 1339.
[28] Rochat, S.; Swagger, T. M. ACS Appl. Mater. Inter. 2013, 5, 4488.
[29] Zhu, C. L.; Liu, L. B.; Yang, Q.; Lv, F. T.; Wang, S. Chem. Rev. 2012, 112, 4687.
[30] Wang, M. F.; Zou, S.; Guerin, G. Macromolecules 2008, 41, 6993
[31] Pu, K. Y.; Li, K.; Liu, B. Adv. Funct. Mater. 2010, 20, 2770.
[32] Zhang, Z. Y.; Lu, X. M.; Fan, Q. L.; Hu, W. B.; Huang, W. Polym. Chem. 2011, 2, 2369.
[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.
[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.
[35] Jiang, R. C.; Lu, X. M.; Yang, M. H.; Deng, W. X.; Fan, Q. L.; Huang, W. Biomacromolecules 2013, 14, 3643.
[36] Zhang, Z. Y. Ph.D. Dissertation, Nanjing University of Posts and Telecommunications, Nanjing, 2014. (张志勇, 博士论文, 南京邮电大学, 南京, 2014.)
[37] Zhang, L. B.; Zhu, J. B.; Li, T.; Wang, E. K. Anal. Chem. 2011, 83, 8871.
[38] Golub, E.; Freeman, R.; Willner, I. Anal. Chem. 2013, 85, 12126.
[39] Lin, Z. H.; Pan, D.; Hu, T. Y.; Liu, Z. P.; Su, X. G. Microchim. Acta 2015, 182, 1933.
[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. (羊小海, 王胜锋, 王柯敏, 罗晓明, 谭蔚泓, 崔亮, 高等学校化学学报, 2009, 30, 899.)
[41] Wang, Y. Y.; Liu, B. Langmuir 2009, 25, 12787.
[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.

基于水溶性共轭聚合物分子刷的高灵敏凝血酶生物传感器

Highly Sensitive Protein Biosensor based on a Conjugated Polymer Brush

PDF

可视化

被引次数

摘要/Abstract

引用此文

分享此文

参考文献

相关文章 15

编辑推荐

相关统计

本文评价

[1]	贾凌轩, 詹泽庞, 贺紫晗, 狄重安, 朱道本. 面向神经电子接口器件的有机材料进展与展望^★[J]. 化学学报, 2023, 81(9): 1175-1186.
[2]	孙博, 琚雯雯, 王涛, 孙晓军, 赵婷, 卢晓梅, 陆峰, 范曲立. 高分散共轭聚合物-金属有机框架纳米立方体的制备及抗肿瘤应用[J]. 化学学报, 2023, 81(7): 757-762.
[3]	李兰英, 陶晴, 闻艳丽, 王乐乐, 郭瑞妍, 刘刚, 左小磊. 多聚腺嘌呤DNA探针及其生物传感应用^★[J]. 化学学报, 2023, 81(6): 681-690.
[4]	朱子煜, 梁阿新, 浩天瑞霖, 唐珊珊, 刘淼, 解炳腾, 罗爱芹. 生物传感器在新冠病毒检测中的应用[J]. 化学学报, 2023, 81(3): 253-263.
[5]	齐子朋, 高冬, 朱志成, 贺志远, 白国英. 聚乙烯醇调控水溶性共轭聚噻吩的光学性质[J]. 化学学报, 2022, 80(7): 921-928.
[6]	王志琴, 项博, 黄晓宇, 陆国林, 冯纯. 磷钨酸对对苯撑乙烯撑寡聚物-b-聚(2-乙烯基吡啶)自晶种行为的影响^※[J]. 化学学报, 2022, 80(3): 297-302.
[7]	浩天瑞霖, 朱子煜, 蔡艳慧, 王微, 王祯, 梁阿新, 罗爱芹. 基于共价有机框架的电化学生物传感器在生物样品检测中的应用[J]. 化学学报, 2022, 80(11): 1524-1535.
[8]	师瑶, 夏乾峰, 何政清, 鞠熀先. 登革热病毒检测的生物传感技术[J]. 化学学报, 2022, 80(1): 69-79.
[9]	廖妮, 钟霞, 梁文斌, 袁若, 卓颖. ECL金属-有机框架(MOF)生物传感平台用于肿瘤细胞分泌H₂O₂的测定[J]. 化学学报, 2021, 79(10): 1257-1264.
[10]	张昊, 黄龙, 李涛, 刘宾, 白泽明, 李小娜, 陆丹. 聚芴类共轭聚合物(PFs)从溶液到薄膜凝聚态演变过程中的定量构效关系[J]. 化学学报, 2019, 77(5): 397-405.
[11]	金鑫, 王晓英. 口腔癌相关唾液肿瘤生物标志物的分析检测研究进展[J]. 化学学报, 2019, 77(4): 340-350.
[12]	刘晔, 袁俊, 邹应萍, 李永舫. 呋喃稠环类共轭有机分子和聚合物研究进展[J]. 化学学报, 2017, 75(3): 257-270.
[13]	林若韵, 陈阳, 陶广宇, 裴晓静, 刘锋, 李娜. 基于G-四链体模板的银纳米簇共振能量转移比率检测miRNA[J]. 化学学报, 2017, 75(11): 1103-1108.
[14]	胡正利, 杜冀晖, 应佚伦, 彭岳一, 曹婵, 龙亿涛. 纳米孔道单分子检测结直肠癌MicroRNAs[J]. 化学学报, 2017, 75(11): 1087-1090.
[15]	朱昊云, 黄威, 黄宇立, 汪伟志. 硅纳米线和聚3-十二烷基噻吩混合薄膜在场效应晶体管中的应用研究[J]. 化学学报, 2016, 74(5): 429-434.