Magnetic gold nanoparticles in SERS-based sandwich immunoassay for antigen detection by well oriented antibodies
Magnetic gold nanoparticles in SERS-based sandwich immunoassay for antigen detection by well oriented antibodies
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DOI：
10.1016/j.bios.2012.12.014
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发表时间：
2013-05-15
影响因子：
12.6
通讯作者：
Ramanaviciene, Almira
中科院分区：
文献类型：
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作者：
Baniukevic, Julija;Boyaci, Ismail Hakki;Ramanaviciene, Almira
本研究旨在建立一种间接、可靠、简便的检测牛白血病病毒抗原gp51的方法。采用表面增强拉曼散射(SERS)作为检测方法。用抗体定向或随机修饰的磁性金纳米颗粒(MNP-Au)与gp51结合。高效液相色谱分析表明，用二硫苏糖醇还原完整抗体后得到的抗体片段具有最好的抗体固定化和抗原捕获效率。为了提高免疫分析的效率和灵敏度，我们构建了由包覆5-硫代-硝基苯甲酸和共价结合抗体的金纳米棒组成的拉曼标记。检测gp51抗原的最低检测限和最低检出限分别为0.95微克/毫升和3.14微克毫克毫升。该免疫分析方法成功地应用于牛奶样品中gp51的快速、可靠和选择性检测。我们相信，所提出的基于SERS的免疫分析模式可以应用于其他蛋白质的检测。(C)2012爱思唯尔B.V.保留所有权利。
The aim of the study was to develop an indirect, robust and simple in application method for the detection of bovine leukemia virus antigen gp51. Surface-enhanced Raman scattering (SERS) was applied as detection method. Magnetic gold nanoparticles (MNP-Au) modified by antibodies in oriented or random manner were used for the binding of gp51. The high performance liquid chromatography analysis indicated that the best antibody immobilization and antigen capturing efficiency was achieved using fragmented antibodies obtained after reduction of intact antibodies with dithiothreitol. In order to increase efficiency and sensitivity of immunoassay Raman labels consisting of gold nanorods coated by 5-thio-nitrobenzoic acid layer with covalently bounded antibodies have been constructed. The LOD and LOQ of the proposed immunoassay for antigen gp51 detection were found to be 0.95 mu g mL(-1) and 3.14 mu g mL(-1), respectively. This immunoassay was successfully applied for the detection of gp51 in milk samples in a rapid, reliable and selective manner. We believe that the proposed SERS-based immunoassay format can be applied for the detection of other proteins. (c) 2012 Elsevier B.V. All rights reserved.