National Institute of Technology, M. G. Avenue, Durgapur 713209, India.
CSIR-Central Mechanical Engineering Research Institute, M. G. Avenue, Durgapur 713209, India.
Bioelectrochemistry. 2017 Dec;118:25-30. doi: 10.1016/j.bioelechem.2017.06.009. Epub 2017 Jun 30.
In this study, an immunosensor based on screen-printed electrode (SPE) has been developed for the detection of Human Epidermal Growth Factor Receptor-2 (HER2) antigen. The SPEs were fabricated and a sandwich enzyme linked immunosorbent assay (ELISA) format was followed for the construction of the immunosensor. The capture antibody (mouse anti-human ErbB2) was coated onto the electrode surface without any prior surface modification, followed by the addition of recombinant human HER2 antigen. Biotinylated goat anti-human ErbB2 was used as the detection antibody which was linked to streptavidin conjugated horseradish peroxidase (HRP). 3,3',5,5'-tetramethylbenzidine (TMB) was used as the substrate. The redox reaction was measured using cyclic voltammetry at scan rate of 50mV/s for the quantification of the antigen in solution. The biotin-avidin chemistry enabled the accurate detection of the antigen in nanogram levels. The amperometric signal obtained increased linearly with increase in the HER2 concentration and two-fold linear range was obtained between 5ng/ml-20ng/ml and 20-200ng/ml respectively. The limit of detection (LOD) and the limit of quantification (LOQ) of this immunosensor were found to be 4ng/ml and 5ng/ml respectively. The detection of HER2 in the serum samples of invasive and non-invasive breast cancer patients has been realized.
在这项研究中，开发了一种基于丝网印刷电极（SPE）的免疫传感器，用于检测人表皮生长因子受体 2（HER2）抗原。制备了 SPE，并采用夹心酶联免疫吸附测定（ELISA）格式构建了免疫传感器。将捕获抗体（鼠抗人 ErbB2）涂覆到电极表面，无需进行任何表面修饰，然后加入重组人 HER2 抗原。生物素化山羊抗人 ErbB2 用作检测抗体，与辣根过氧化物酶（HRP）偶联的链霉亲和素结合。使用 3,3',5,5'-四甲基联苯胺（TMB）作为底物。使用循环伏安法在扫描速率为 50mV/s 下测量氧化还原反应，以定量溶液中的抗原。生物素-亲和素化学可实现抗原在纳克水平的准确检测。获得的电流信号随 HER2 浓度的增加呈线性增加，分别在 5ng/ml-20ng/ml 和 20-200ng/ml 之间获得两倍线性范围。该免疫传感器的检测限（LOD）和定量限（LOQ）分别为 4ng/ml 和 5ng/ml。已经实现了对浸润性和非浸润性乳腺癌患者血清样本中 HER2 的检测。
