基于主客体竞争模式的凝血酶适配体传感器的研究
|
摘要
基于β-环糊精(β-CD)主客体竞争模式,构建了开关型凝血酶适配体电化学传感器。将末端修饰了二茂铁(Fc)的核酸适配体通过与β-CD的主客体识别固定在金电极表面,当凝血酶存在时,适配体由原来的直立线状构型变为"G-四链体",远离电极表面,适配体探针的氧化还原电流强度减小,即"Signal-off"。利用此效应对凝血酶进行了灵敏检测,结果表明,在5.0×10~(-13)~5.0×10~(-9)mol/L浓度范围内,凝血酶的浓度与电化学响应信号呈良好的线性关系,检出限为2.0×10~(-13)mol/L(3σ)。与其它蛋白分子相比,本方法对凝血酶蛋白的检测具有高特异性。本传感器构建简单,再生性好,为生物血清样本中凝血酶的实时高效检测提供了方法。
A switch thrombin aptamer sensor was constructed based on the host-guest competition mode ofβ-cyclodextrin(β-CD).The aptamer that modified with ferrocene(Fc) at its terminal was fixed on the surface of the gold electrode via the host-guest recognization with β-CD.When thrombin was present,the configuration of aptamer transformed from vertical linear to "G-quadruplex"and far away from the electrode surface,which resulted in a decrease in the redox current of the aptamer probe and produce " Signal-off" effect.On the basis of this,a high sensitive detection of thrombin was made.The detection result indicated that the thrombin concentration had a consideration linear response to the signal of aptasensor between 5.0×10~(-13)-5.0×~(-9)mol/L,and as low as 2.0×10~(-13)mol/L thrombin had been detected.This method for thrombin detection showed a higher specificity compared to other protein molecules.Besides,the sensor was constructed easily and possessed excellent regeneration,which provided a significant platform for highly efficient real-time detection of thrombin in biological serum samples.
A switch thrombin aptamer sensor was constructed based on the host-guest competition mode ofβ-cyclodextrin(β-CD).The aptamer that modified with ferrocene(Fc) at its terminal was fixed on the surface of the gold electrode via the host-guest recognization with β-CD.When thrombin was present,the configuration of aptamer transformed from vertical linear to "G-quadruplex"and far away from the electrode surface,which resulted in a decrease in the redox current of the aptamer probe and produce " Signal-off" effect.On the basis of this,a high sensitive detection of thrombin was made.The detection result indicated that the thrombin concentration had a consideration linear response to the signal of aptasensor between 5.0×10~(-13)-5.0×~(-9)mol/L,and as low as 2.0×10~(-13)mol/L thrombin had been detected.This method for thrombin detection showed a higher specificity compared to other protein molecules.Besides,the sensor was constructed easily and possessed excellent regeneration,which provided a significant platform for highly efficient real-time detection of thrombin in biological serum samples.
引文
1 Archiniegas E,Neves C Y,Candelle D,Cardier,J E.DNA Cell Biol.,2004,23(12):815-825
2 Popovic M,Smiljanic K,Dobutovic B,Syrovets T,Simmet T,Lsenovic E R.Mol.Cell.Biochem.,2012,359(1-2):301-313
3 MENG Yu-Hong,YU Ji-Yao,LU Ying-Lin.Journal of Experimental Hematology.,2007,(03):671-674孟宇宏,虞积耀,陆应麟.中国实验血液学杂志,2007,(03):671-674
4 Hu L,Lee M,Campbell W,Perez-Soler R,Karpatkin S.Blood,2004,104(9):2746-2751
5 Nierodzik M L,Karpatkin S.Cancer Cell,2006,10(5):355-362
6 Huang K J,Liu Y J,Cao J T,Wang H B.RSC Adv.,2014,4(69):36742-36748
7 Wu Z S,Zheng F,Shen G L,Yu R Q.Biomaterials,2009,30(15):2950-2955
8 Cui H F,Cheng L,Zhang J,Liu R H,Zhang C,Fan H.Biosens.Bioelectron.,2014,56:124-128
9 Hong N,Cheng L,Wei G B,Chen C D,He L L,Kong D R,Zeng J X,Cui H F,Fan H.Biosens.Bioelectron.,2017,91:110-114
10 CUI Cheng,JIANG Yu-Yang.Chinese Journal of New Drugs.,2011,(5):407-410崔承,蒋宇扬.中国新药杂志,2011,(5):407-410
11 LIU Zhen-Bao,SHI Yi-Qian,CHEN Zhang-Ren,DUAN Lin-Hua,WANG Xin-Ru.Chinese Science Bulletin.,2014,(14):1267-1279刘珍宝,石依倩,陈长仁,段林花,王薪茹.科学通报,2014,(14):1267-1279
12 WANG Yi-Xian,YE Zun-Zhong,SI Cheng-Yan,YING Yi-Bin.Chinese J.Anal.Chem.,2012,40(4):634-642王一娴,叶尊忠,斯城燕,应义斌.分析化学,2012,40(4):634-642
13 Tian R,Chen X J,Li Q W,Yao C.Anal.Biochem.,2016,500:73-79
14 ZHANG Song-Bai,ZHENG Li-Ying,HU Xia,SHEN Guang-Yu,LIU Xue-Wen,SHEN Guo-Li,YU Ru-Qin.Chinese J.Anal.Chem.,2015,43(11):1688-1694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(11):1688-1694
15 Cheng L,Zhang J,Lin Y,Wang Q,Zhang X X,Ding Y H,Cui H F,Fan H.Anal Biochem.,2015,491:31-36
16 Yang S M,Li H,Zha W L,Sun Q,Zheng L Z,Chen A X.J.Solid State Electrochem.,2013,17(10):2603-2610
17 Cai H,Lee T,Hsing I M.Sens.Actuators B,2006,114(1):433-437
18 He C,Xu Z H,Sun T,Wang L.Appl.Biochem.Biotechnol.,2014,172(2):1018-1026
19 Zhao J,Zhang Y Y,Li H T,Wen Y Q,Fan X Y,Lin F B,Tan L,Yao S Z.Biosens.Bioelectron.,2011,26(5):2297-2303
20 Gao F L,Du L L,Zhang Y,Zhou F Y,Tang D Q.Biosens.Bioelectron.,2016,86:185-193
21 Wang Y G,Zhang Y,Yan T,Fan D W,Du B,Ma H M,Wei Q.Biosens.Bioelectron.,2016,80:640-646
22 Radi A E,Sanchez J,Baldrich E,O'Sullivan,C K.J.Am.Chem.Soc.,2006,128(1):117-124
23 Mir M,Katakis I.Mol.Bio Sys.,2007,3(9):620-622
24 Xiao Y,Piorek B D,Plaxco K W,Heeger A J.J.Am.Chem.Soc.,2005,127(51):17990-17991
25 SHEN Hai-Min,JI Hong-Bing,WU Hong-Ke,SHI Hong-Xin.Chinese Journal of Organic Chemistry.,2014,34(8):1549-1572沈海民,纪红兵,武宏科,史鸿鑫.有机化学,2014,34(8):1549-1572
26 Ha W,Kang Y,Peng S L,Ding L S,Zhang S,Li B J.Nanotechnology,2013,24(49):495103
2 Popovic M,Smiljanic K,Dobutovic B,Syrovets T,Simmet T,Lsenovic E R.Mol.Cell.Biochem.,2012,359(1-2):301-313
3 MENG Yu-Hong,YU Ji-Yao,LU Ying-Lin.Journal of Experimental Hematology.,2007,(03):671-674孟宇宏,虞积耀,陆应麟.中国实验血液学杂志,2007,(03):671-674
4 Hu L,Lee M,Campbell W,Perez-Soler R,Karpatkin S.Blood,2004,104(9):2746-2751
5 Nierodzik M L,Karpatkin S.Cancer Cell,2006,10(5):355-362
6 Huang K J,Liu Y J,Cao J T,Wang H B.RSC Adv.,2014,4(69):36742-36748
7 Wu Z S,Zheng F,Shen G L,Yu R Q.Biomaterials,2009,30(15):2950-2955
8 Cui H F,Cheng L,Zhang J,Liu R H,Zhang C,Fan H.Biosens.Bioelectron.,2014,56:124-128
9 Hong N,Cheng L,Wei G B,Chen C D,He L L,Kong D R,Zeng J X,Cui H F,Fan H.Biosens.Bioelectron.,2017,91:110-114
10 CUI Cheng,JIANG Yu-Yang.Chinese Journal of New Drugs.,2011,(5):407-410崔承,蒋宇扬.中国新药杂志,2011,(5):407-410
11 LIU Zhen-Bao,SHI Yi-Qian,CHEN Zhang-Ren,DUAN Lin-Hua,WANG Xin-Ru.Chinese Science Bulletin.,2014,(14):1267-1279刘珍宝,石依倩,陈长仁,段林花,王薪茹.科学通报,2014,(14):1267-1279
12 WANG Yi-Xian,YE Zun-Zhong,SI Cheng-Yan,YING Yi-Bin.Chinese J.Anal.Chem.,2012,40(4):634-642王一娴,叶尊忠,斯城燕,应义斌.分析化学,2012,40(4):634-642
13 Tian R,Chen X J,Li Q W,Yao C.Anal.Biochem.,2016,500:73-79
14 ZHANG Song-Bai,ZHENG Li-Ying,HU Xia,SHEN Guang-Yu,LIU Xue-Wen,SHEN Guo-Li,YU Ru-Qin.Chinese J.Anal.Chem.,2015,43(11):1688-1694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(11):1688-1694
15 Cheng L,Zhang J,Lin Y,Wang Q,Zhang X X,Ding Y H,Cui H F,Fan H.Anal Biochem.,2015,491:31-36
16 Yang S M,Li H,Zha W L,Sun Q,Zheng L Z,Chen A X.J.Solid State Electrochem.,2013,17(10):2603-2610
17 Cai H,Lee T,Hsing I M.Sens.Actuators B,2006,114(1):433-437
18 He C,Xu Z H,Sun T,Wang L.Appl.Biochem.Biotechnol.,2014,172(2):1018-1026
19 Zhao J,Zhang Y Y,Li H T,Wen Y Q,Fan X Y,Lin F B,Tan L,Yao S Z.Biosens.Bioelectron.,2011,26(5):2297-2303
20 Gao F L,Du L L,Zhang Y,Zhou F Y,Tang D Q.Biosens.Bioelectron.,2016,86:185-193
21 Wang Y G,Zhang Y,Yan T,Fan D W,Du B,Ma H M,Wei Q.Biosens.Bioelectron.,2016,80:640-646
22 Radi A E,Sanchez J,Baldrich E,O'Sullivan,C K.J.Am.Chem.Soc.,2006,128(1):117-124
23 Mir M,Katakis I.Mol.Bio Sys.,2007,3(9):620-622
24 Xiao Y,Piorek B D,Plaxco K W,Heeger A J.J.Am.Chem.Soc.,2005,127(51):17990-17991
25 SHEN Hai-Min,JI Hong-Bing,WU Hong-Ke,SHI Hong-Xin.Chinese Journal of Organic Chemistry.,2014,34(8):1549-1572沈海民,纪红兵,武宏科,史鸿鑫.有机化学,2014,34(8):1549-1572
26 Ha W,Kang Y,Peng S L,Ding L S,Zhang S,Li B J.Nanotechnology,2013,24(49):495103