Improved procedures for immobilisation of oligonucleotides on gold-coated piezoelectric quartz crystals
Improved procedures for immobilisation of oligonucleotides on gold-coated piezoelectric quartz crystals
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DOI：
10.1016/s0956-5663(02)00084-2
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发表时间：
2002-12-01
影响因子：
12.6
通讯作者：
Turner, APF
中科院分区：
文献类型：
--
作者：
Tombelli, S;Mascini, M;Turner, APF
DNA杂交技术的高灵敏度和特异性使其成为环境或临床分析的有力工具。本工作描述了一种用于检测杂交反应的DNA压电生物传感器的发展。注意力集中在可用于将寡核苷酸固定到石英晶体的金表面上的涂层化学的选择上。考虑到固定化探针的量、杂交反应的程度、再生的可能性和不存在非特异性吸附，对四种固定化程序进行了测试和比较。所有实验均用25个碱基的寡核苷酸（探针、靶和非互补寡核苷酸）进行。这四种涂层方法都是基于使用自组装单分子膜（SAM）。其中三个采用链霉亲和素和生物素之间的相互作用的生物素化探针的固定。结果表明，生物素化的探针在链霉亲和素上的固定化连接到羧化葡聚糖层，提供了更高的灵敏度检测的杂交反应，非特异性吸附的情况下，相对于再生步骤和更高的稳定性。(C)2002 Elsevier Science B.V.保留所有权利。
The high sensitivity and specificity of DNA hybridisation techniques makes them powerful tools for environmental or clinical analysis. This work describes the development of a DNA piezoelectric biosensor for the detection of the hybridisation reaction. Attention was focused on the choice of the coating chemistry that could be used for the immobilisation of oligonucleotides onto the gold surface of the quartz crystal. Four immobilisation procedures were tested and compared considering the amount of immobilised probe, the extent of the hybridisation reaction, the possibility of regeneration and the absence of non-specific adsorption. All the experiments were performed with oligonucleotides of 25 bases (probe, target and non-complementary oligonucleotide). The four coating methods were all based on the use of self-assembled monolayers (SAM). Three of them employed the interaction between streptavidin and biotin for the immobilisation of a biotinylated probe. Results indicated that immobilisation of a biotinylated probe on streptavidin linked to a layer of carboxylated dextran provides higher sensitivity for the detection of the hybridisation reaction, absence of non-specific adsorption and a higher stability with respect to the regeneration step. (C) 2002 Elsevier Science B.V. All rights reserved.