Surface plasmon resonance immunosensor for highly sensitive detection of 2,4,6-trinitrotoluene
Surface plasmon resonance immunosensor for highly sensitive detection of 2,4,6-trinitrotoluene
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DOI：
10.1016/j.bios.2004.06.044
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发表时间：
2005-03-15
影响因子：
12.6
通讯作者：
Miura, N
中科院分区：
文献类型：
--
作者：
Shankaran, DR;Gobi, KV;Miura, N
利用2，4，6-三硝基苯酚-卵清蛋白（TNP-OVA）偶联物与抗2，4，6-三硝基苯酚抗体（抗TNP抗体）的免疫反应，研究了表面等离子体共振（SPR）免疫分析检测2，4，6-三硝基甲苯（TNT）的传感特性。TNP-OVA偶联物通过物理固定的方式附着到SPR-金传感表面，其经历与抗TNP抗体的结合相互作用。固定化和结合过程进行了研究，从SPR共振角度的变化。TNT的定量基于间接竞争免疫测定的原理，其中在溶液中存在游离TNT的情况下，TNP-OVA缀合物和抗TNP抗体之间的免疫反应被抑制。共振角位移的减小与用于孵育的TNT浓度的增加成比例。该方法在0.09 ~ 1000 ng/ml的浓度范围内具有良好的灵敏度和稳定性。该免疫传感器可以检测低至0.09 ng/ml的TNT，响应时间约为22 min。通过胃蛋白酶溶液的短暂流动使传感器表面再生，该溶液破坏抗原-抗体复合物而不破坏缀合物生物膜。研究了SPR传感器对一些结构相关的硝基芳香族化合物的交叉反应性以及在这些硝基芳香族化合物存在下对TNT的检测。SPR传感器对2，4-二硝基甲苯（2，4-DNT）、1，3-二硝基苯（1，3-DNB）、2-氨基-4，6-二硝基甲苯（2A-4，6-DNT）和4-氨基-2，6-二硝基甲苯（4A-2，6-DNT）的交叉反应性很低（≤ 1. 1%）。所提出的免疫传感器的分析特性是非常有前途的新的现场便携式传感器的现场探测地雷的发展。(C)2004年爱思唯尔B。V.保留所有权利。
We have examined the sensing characteristics of a surface plasmon resonance (SPR) immunoassay for the detection of 2,4,6-trinitrotoluene (TNT) using an immunoreaction between 2,4,6-trinitrophenol-ovalbumin (TNP-OVA) conjugate and anti-2,4,6-trinitrophenol antibody (antiTNP antibody). TNP-OVA conjugate was attached to a SPR-gold sensing surface by means of physical immobilization, which undergoes binding interaction with anti-TNP antibody. Both the immobilization and binding processes were studied from a change in the SPR-resonance angle. The quantification of TNT is based on the principle of indirect competitive immunoassay, in which the immunoreaction between the TNP-OVA conjugate and anti-TNP antibody was inhibited in the presence of free TNT in solution. The decrease in the resonance angle shift is proportional to an increase in concentration of TNT used for incubation. The immunoassay exhibited excellent sensitivity for the detection of TNT in the concentration range from 0.09 to 1000 ng/ml with good stability and reproducibility. The immunosensor developed could detect TNT as low as 0.09 ng/ml, within a response time of similar to22 min. The sensor surface was regenerated by a brief flow of pepsin solution, which disrupts the antigen-antibody complex without destroying the conjugate biofilm. Cross-reactivity of the SPR sensor to some structurally related nitroaromatic derivative and the detection of TNT in the presence of these nitroaromatic compounds were investigated. The cross-reactivity of the SPR sensor to 2,4-dinitrotoluene (2,4-DNT), 1,3-dinitrobenzene (1,3-DNB), 2-amino-4,6-dinitrotoluene (2A-4,6-DNT) and 4-amino-2,6-dinitrotoluene (4A-2,6-DNT) were very low (less than or equal to 1. 1%). The analytical characteristics of the proposed immunosensor are highly promising for the development of new field-portable sensors for on-site detection of landmines. (C) 2004 Elsevier B. V. All rights reserved.