Characterisation of an antibody coated microcantilever as a potential immuno-based biosensor
Characterisation of an antibody coated microcantilever as a potential immuno-based biosensor
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DOI：
10.1016/s0956-5663(01)00276-7
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发表时间：
2002-03-01
影响因子：
12.6
通讯作者：
Leech, D
中科院分区：
文献类型：
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作者：
Grogan, C;Raiteri, R;Leech, D
在这项研究中，我们研究了肌红蛋白单克隆抗体共价固定在微制造悬臂表面上的活性、稳定性、寿命和可重用性。这些传感表面是我们感兴趣的新型悬臂免疫传感器的发展。对于这种传感器，抗体层代表传感元件，而微悬臂充当机械换能器。一种生产可重复使用的生物涂层的方法已经用不同的独立技术进行了测试。酶联免疫吸附试验（ELISA）用于确定活性抗体包被的存在，并监测固定抗体的寿命和稳定性。通过这一分析，固定化抗体层的活性被发现随着蔗糖作为稳定剂的引入而更加稳定。每个再生步骤后，将蔗糖涂在固定抗体层上。发现固定抗体具有长达7周的稳定活性寿命。荧光显微镜被用来提供关于涂层在金和氮化硅侧的分布的信息。原子力显微镜用于确定生物涂层在悬臂上的存在，并获得传感器生物元件表面形态的信息。综合结果为优化传感元件的开发提供了有价值的信息，并展示了一套用于未来传感器到传感器表征的方法。初步实验结果显示，基于微悬臂的传感器原型检测到针对肌红蛋白的抗体活性，证实了所提出的免疫传感技术的动机和潜力。(C) 2002 Elsevier Science B.V.版权所有
In this study, we investigated the activity, stability, lifetime and re-usability of monoclonal antibodies to myoglobin covalently immobilised onto microfabricated cantilever surfaces. These sensing surfaces are of interest to us in the development of novel cantilever-based immunosensors. For such sensors the antibody layer represents the sensing element while the microcantilever acts as a mechanical transducer. A procedure for producing re-usable biological coatings has been tested with different independent techniques. An Enzyme Linked Immunosorbent Assay (ELISA) was used to determine the presence of an active antibody coating, and to monitor the lifetime and stability of the immobilised antibody. Through this analysis, the activity of the immobilised antibody layer was found to be more stable with the introduction of sucrose, as a stabilising agent. Sucrose was applied to the immobilised antibody layer after each regeneration step. The immobilised antibody was found to have a stable active lifetime for up to 7 weeks. Fluorescence microscopy was used to give information on the distribution of the coating on the gold and silicon nitride sides of the cantilever. Atomic Force Microscopy was used to determine the presence of the biological coating on the cantilever and to obtain information on the surface morphology of the biological element of the sensor. The combined results provide valuable information on the development of an optimised sensing element and demonstrate a set of methods to use for future sensor-to-sensor characterisation. Preliminary experimental results showing the antibody activity against myoglobin, detected with a microcantilever based sensor prototype confirmed the motivations and potentialities of the proposed immunosensing technique. (C) 2002 Elsevier Science B.V. All rights reserved.