Label free gravimetric detection of epidermal growth factor receptor by antibody immobilization on quartz crystal microbalance
Label free gravimetric detection of epidermal growth factor receptor by antibody immobilization on quartz crystal microbalance
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DOI：
10.1016/j.procbio.2010.10.006
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发表时间：
2011-02-01
影响因子：
4.4
通讯作者：
Lin, Feng-Huei
中科院分区：
文献类型：
--
作者：
Chen, Jung-Chih;Sadhasivam, S.;Lin, Feng-Huei
建立了一种高效、简便的检测表皮生长因子受体（EGFR）的方法。通过硫代水杨酸的自组装单层膜（SAM）和1-[3-（二甲氨基）丙基]-3-乙基碳二亚胺盐酸盐（EDC）和N-羟基琥珀酰亚胺（NHS）的活化，将抗EGFR抗体固定在QCM电极的金表面。为了评估QCM的灵敏度，研究了两种分析方法，即浸渍干燥法和流动池法。尝试了两种不同的抗体固定模式，如化学交联和通过蛋白G的位点特异性。因此，蛋白G介导的抗体固定化由于其对抗原的高亲和力而控制电极表面上的抗体分子的量和取向。因此，将类似的固定策略用于EGFR的实时结合的流动池分析。在频率偏移和不同EGFR浓度（0.01 - 10 μ g/mL）之间观察到线性关系。用石英晶体微天平对免疫传感器的分步组装进行了表征。还证明，制备的传感器表面足够稳定，能够承受用0.1 M NaOH反复进行的表面再生。(C)2010爱思唯尔有限公司保留所有权利。
An efficient and simple method for the detection of epidermal growth factor receptor (EGFR) using anti-EGFR immobilized quartz crystal microbalance (QCM) was developed. The immobilization of anti-EGFR on the gold surface of QCM electrode was constructed by a self-assembled monolayer (SAM) of thiosalicylic acid followed by the activation of 1-[3-(dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDC) and N-hydroxysuccinimide (NHS). In order to assess the sensitivity of QCM, two analytical procedures namely, dip-and-dry and flow cell methods were investigated. Two different antibody immobilization patterns such as chemical cross-linking and site-specific via protein G were attempted. Accordingly, protein G mediated antibody immobilization controls the quantity and orientation of the antibody molecules on the electrode surface for its high affinity to antigens. Thus, a similar immobilization strategy was utilized for the flow-cell analysis of real-time binding of EGFR. A linear relationship was observed between the frequency shift and different EGFR concentrations (0.01-10 mu g mL(-1)). The stepwise assembly of the immunosensor was characterized by quartz crystal microbalance. It was also demonstrated that, the prepared sensor surface was stable enough to withstand the repeated surface regeneration with 0.1 M NaOH. (C) 2010 Elsevier Ltd. All rights reserved.