Evaluation of intact- and fragmented-antibody based immunosensors by total internal reflection ellipsometry
Evaluation of intact- and fragmented-antibody based immunosensors by total internal reflection ellipsometry
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DOI：
10.1016/j.snb.2011.08.029
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发表时间：
2011-12-15
影响因子：
8.4
通讯作者：
Ramanavicius, Arunas
中科院分区：
文献类型：
--
作者：
Balevicius, Zigmas;Ramanaviciene, Almira;Ramanavicius, Arunas
利用全内反射椭圆偏振（TIRE）技术研究免疫传感器的生物识别层，以估计固定的完整抗体和片段抗体的取向。研究了在金底物上形成的两种不同制备的生物识别层：一层是基于完整抗体（integrity - ab），第二层是基于化学碎片化的特异性抗体（fragment - ab），通过还原完整抗体（integrity - ab）得到。结果表明，TIRE能够分辨完整ab层和碎片ab层的纳米结构差异。应用多层模型进行计算，结果表明，固定过程后，碎片化抗体活性位点与金表面之间的距离小于碎片化抗体的理论维数。此外，计算出TIRE的Delta（lambda）参数的分析灵敏度是Psi（lambda）参数的灵敏度的5.89倍，这实际上与基于表面等离子体共振（SPR）的免疫传感器的灵敏度相似。(C) 2011 Elsevier B.V.版权所有
Total internal reflection ellipsometry (TIRE) technique was used to investigate biological recognition layers of immunosensors in order to estimate orientation of immobilized intact- and fragmented-antibodies. Two differently prepared biological recognition layers formed on gold-substrate were investigated: one layer was based on intact-antibodies (intact-Ab), second layer was based on chemically fragmented specific-antibodies (frag-Ab), which were obtained by reduction of intact-Ab. It was shown that TIRE enables to resolve differences in nanostructures of intact-Ab and frag-Ab layers. A multilayer model applied in our calculations shows that the distance between fragmented-antibody active sites and gold-surface after the immobilization process is lower than theoretical dimension of fragmented-antibodies. Moreover, it was calculated that analytical sensitivity of the Delta(lambda) parameter of TIRE was 5.89 times better if compared to the sensitivity of the Psi(lambda) parameter, which is in fact similar to the sensitivity of surface plasmon resonance (SPR) based immunosensors. (C) 2011 Elsevier B.V. All rights reserved.