基于Mn掺杂ZnS量子点磷光内滤效应检测β-葡萄糖醛酸酶
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摘要
高效荧光内滤分析的关键是使猝灭剂的吸收峰与荧光团的激发峰或发射峰最大限度地重叠。本研究将Mn掺杂Zn S量子点(Mn-Zn S QDs)作为内滤体系的荧光体,4-硝基苯-β-D-葡糖苷酸(PNPG)作为吸收体,实现了β-葡萄糖醛酸酶(GUS)的特异性检测。PNPG的吸收光谱与Mn-Zn S QDs的激发光谱大幅重叠,能够高效猝灭Mn-Zn S QDs的磷光。由于Mn-Zn S QDs具有较大的斯托克斯位移(约300 nm),其激发光谱和发射光谱与GUS的酶解产物PNP的吸收光谱几乎无重叠,因而实现了GUS的磷光Turn-on检测。此外,Mn-Zn S QDs具有优异的室温磷光性质,可以避开生物组织荧光背景,从而可有效应用于生物样品分析。据此建立了一种基于内滤效应的GUS磷光探针,实现了对大肠杆菌的测定。本方法在最优的实验条件下对10~300 U/L的GUS有线性响应,检出限为7 U/L,且本策略相对于紫外-可见光谱法有很好的抗基底干扰能力。
The key to maximize the sensitivity of inner filter effect( IFE)-based assay is to enlarge the overlap between the absorption spectra of the absorber and the excitation or emission spectra of the fluorophore. In this work,Mn-doped Zn S quantum dots( QDs) were chosen for IFE-based detection of β-glucuronidase( GUS),since the excitation of QDs was perfectly overlapped with the absorption of the substrate of GUS,namely 4-nitrophenyl-β-D-glucuronide( PNPG). In addition,the phosphorescence emission from Mn-doped ZnS QDs could eliminate the fluorescence background from biological samples. Therefore, a simple turn-on phosphorescent GUS assay was developed,with the linear range of 10-300 U/L and limit of detection of 7 U/L( S/N = 3).
The key to maximize the sensitivity of inner filter effect( IFE)-based assay is to enlarge the overlap between the absorption spectra of the absorber and the excitation or emission spectra of the fluorophore. In this work,Mn-doped Zn S quantum dots( QDs) were chosen for IFE-based detection of β-glucuronidase( GUS),since the excitation of QDs was perfectly overlapped with the absorption of the substrate of GUS,namely 4-nitrophenyl-β-D-glucuronide( PNPG). In addition,the phosphorescence emission from Mn-doped ZnS QDs could eliminate the fluorescence background from biological samples. Therefore, a simple turn-on phosphorescent GUS assay was developed,with the linear range of 10-300 U/L and limit of detection of 7 U/L( S/N = 3).
引文
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2 Bosslet K,Czech J,Hoffmann D.Tumor Targeting,1995,1:45-50
3 Bosslet K,Straub R,Blumrich M,Czech J,Gerken M,Sperker B,Kroemer H K,Gesson J P,Koch M,Monneret C.Cancer Res.,1998,58(6):1195-1201
4 Bohnenstengel F,Kroemer H K,Sperker B.J.Chromatogr.B,1999,721(2):295-299
5 Waszkiewicz N,Szajda S D,Konarzewska-Duchnowska E,Zalewska-Szajda B,Gazkowski R,Sawko A,Nammous H,Buko V,Szulc A,Zwierz K,Ladny J R.Postepy Hig.Med.Dosw.,2015,69:436-439
6 Rochelet M,Solanas S,Betelli L,Chantemesse B,Vienney F,Hartmann A.Anal.Chim.Acta,2015,892:160-166
7 TANG Hui,ZHANG Wen,GENG Ping,JIN Li-Tong,LOU Min,WU Zi-Rong.Chinese J.Anal.Chem.,2006,34(6):829-831唐辉,张文,耿萍,金利通,楼旻,吴自荣.分析化学,2006,34(6):829-831
8 Briciu-Burghina C,Heery B,Regan F.Analyst,2015,140(17):5953-5964
9 Yuan L,Lin W,Zheng K,Zhu S.Accounts Chem.Res.,2013,46(7):1462-1473
10 Daly B,Ling J,de Silva A P.Chem.Soc.Rev.,2015,44(13):4203-4211
11 Shao N,Zhang Y,Cheung S,Yang R,Chan W,Mo T,Li K,Liu F.Anal.Chem.,2005,77(22):7294-7303
12 Chen H,Ren J.Talanta,2012,99:404-408
13 Xu Y L,Niu X Y,Chen H L,Zhao S G,Chen X G.Chin.Chem.Lett.,2017,28(2):338-344
14 Yan X,Li H,Han X,Su X.Biosens.Bioelectron.,2015,74:277-283
15 Tang Y,Liu Y,Cao A.Anal.Chem.,2013,85(2):825-830
16 Zhao D,Chen C,Sun J,Yang X.Analyst,2016,141(11):3280-3288
17 Li G,Fu H,Chen X,Gong P,Chen G,Xia L,Wang H,You J,Wu Y.Anal.Chem.,2016,88(5):2720-2726
18 Zhang J Y,Lu X M,Lei Y,Hou X D,Wu P.Nanoscale,2017,40(9):15606-15611
19 Zhang J Y,Zhu A R,Zhao T,Wu L,Wu P,Hou X D.J.Mater.Chem.B,2015,3(29):5942-5950
20 Nikolic D B,Samardzic J T,Bratic A M,Radin I P,Gavrilovic S P,Rausch T,Maksimovic V R.J.Agr.Food Chem.,2010,58(6):3488-3494
21 Zou S,Liu G,Kaleem I,Li C.Process Biochem.,2013,48(2):358-363