基于AgNCs-Ce:ZONPs发光复合纳米材料的电致化学发光免疫传感器对肿瘤标志物灵敏检测的研究
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摘要
该研究以含有铈的氧化锌复合纳米材料(Ce:ZONPs)为固载基质,以BSA作为模板和还原剂在Ce:ZONPs表面原位合成银纳米簇(Ag NCs)制得发光复合纳米材料(Ag NCs-Ce:ZONPs),并以此作为信号探针构建了电致化学发光(Electrochemiluminescence,ECL)免疫传感器用于癌胚抗原(CEA)的灵敏检测。在该工作中,BSA作为天然蛋白质,表面含有大量的氨基和巯基,介于共价键Ag-N和Ag-S,BSA可以作为优良的纳米簇配体。同时通过BSA的成膜性,制备BSA功能化的Ce:ZONPs,从而在Ce:ZONPs@BSA的表面生成大量、粒径小于2 nm的Ag NCs,获得高效、稳定的电致化学发光。最后利用该复合物上BSA剩余的氨基与anti-CEA(Ab2)中的羧基交联,从而制备出Ag NCs-Ce:ZONPs/Ab2探针。电极敏感界面的构建是在裸玻碳电极上修饰一层功能化BSA的TiO_2纳米复合物(TiO_2@BSA),提供充足的活性位点来固载anti-CEA(Ab1),从而制得了ECL免疫传感器。基于夹心型反应模式,此免疫传感器实现了对CEA的灵敏检测,线性范围2.5ng/m L至80 ng/m L。
In this work, the electrochemiluminescent nanocomposite was proposed through Ag NCs in situ generating on the surface of bovine serum albumin(BSA) functionalized zinc oxide containing cerium. Notably, BSA as stabilizing agent provided abundant binding sites to potentially bind and further reduce Ag ions for generating Ag NCs.The Ag NCs-TiO_2 NFs composites achieved highly efficient ECL, biocompatibility, ease of labeling and easy film-forming. In view of these advantages, the composites could have great potential as a signal probe to construct electrochemiluminescence(ECL) immunesensor for detection of carcinoembryonic antigen(CEA). Meanwhile, the nanocomposites(TiO_2@BSA) were modified onto the surface of a glassy carbon electrode(GCE), which afforded a large surface area for anchoring massive Ab1 via CO-NH bonding. Based on the "sandwich reaction" strategy, this immunesensor provided an ultrasensitive ECL detection of CEA with a linear response from 2.5 ng/m L to 80 ng/m L.
In this work, the electrochemiluminescent nanocomposite was proposed through Ag NCs in situ generating on the surface of bovine serum albumin(BSA) functionalized zinc oxide containing cerium. Notably, BSA as stabilizing agent provided abundant binding sites to potentially bind and further reduce Ag ions for generating Ag NCs.The Ag NCs-TiO_2 NFs composites achieved highly efficient ECL, biocompatibility, ease of labeling and easy film-forming. In view of these advantages, the composites could have great potential as a signal probe to construct electrochemiluminescence(ECL) immunesensor for detection of carcinoembryonic antigen(CEA). Meanwhile, the nanocomposites(TiO_2@BSA) were modified onto the surface of a glassy carbon electrode(GCE), which afforded a large surface area for anchoring massive Ab1 via CO-NH bonding. Based on the "sandwich reaction" strategy, this immunesensor provided an ultrasensitive ECL detection of CEA with a linear response from 2.5 ng/m L to 80 ng/m L.
引文
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[2]李偶连,刘翠,童艳丽,等.癌胚抗原(CEA)检测技术的研究进展[J].综述,2008,2:48-51.
[3]杨霞,袁若,柴雅琴,等.基于纳米金/功能化壳聚糖生物复合膜修饰的癌胚抗原免疫传感器的研究[J].数理科学与化学,2008,7:57-61.
[4]张青云,孙丽,张书耕,等.时间分辨荧光免疫法与电化学发光法检测结直肠肿瘤标志CEA的比较[J].世界华人消化杂志,2005,23:2799-2802.
[5]梁臻龙,王策,李炯,等.基于微流控技术与酶促化学发光免疫技术检测血清CEA方法的建立及初步应用评价[J].临床检验技术研究,2015,12:892-894.
[6]姚少利.胶体金法与增强化学发光免疫分析法检测CEA在中老年体检中的应用价值[J].卫生检验,2015,2:127-129.
[7]王晓英.基于电致化学发光技术的纳米生物传感器的设计和研究[D].上海:华东师范大学,2008.
[8]朱晓燕.基于联吡啶钌的电致化学发光新方法研究[D].南昌:南昌大学,2011.
[9]饶海英.三联吡啶钌电化学发光在药物分析中的应用研究[D].江苏:苏州大学,2014.
[10]邵晓东,李瑛.鲁米诺化学发光分析法研究进展[J].化学研究,2019,1:103-112.
[11]顾仁敖,姚建林.贵金属纳米结构和电极的表面性质及SERS研究[D].江苏:苏州大学,2011.
[12]Zongyin Yang,Jinyou Xu,Pan Wang,et al.On-Nanowire Spatial Band Gap Design for White Light Emission[J].Nano Lett,2011,11(11):5085-5089.
[13]Tosatti S,Michel R,Textor M,et al.Self-Assembled Monolayers of Dodecyl and Hydroxy-dodecyl Phosphates on Both Smooth and Rough Titanium and Titanium Oxide Surfaces[J].Langmuir,2002,18:3537-3548.
[14]张普.银、金纳米团簇的合成及其在生化分析中的应用研究[D].重庆:西南大学,2014.
[15]宋琳.Ti O2的形貌控制合成及其催化性质研究[D].哈尔滨:哈尔滨师范大学,2014.