Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics & Molecular Imaging Key Laboratory, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, PR China.
Department of Clinical Laboratory, Wuhan Commercial Staff Hospital, Wuhan 430021, PR China.
Biosens Bioelectron. 2015 Feb 15;64:493-8. doi: 10.1016/j.bios.2014.09.050. Epub 2014 Sep 28.
A new strategy is proposed for determination of carcino-embryonic antigen (CEA) based on aptamer/graphene oxide (Apt/GO) by capillary electrophoresis-chemiluminescence (CE-CL) detection system. CEA aptamer conjugated with horseradish peroxidase (HRP) firstly mixes with GO, and the CL will be quenched because the stack of HRP-Apt on GO leads to chemiluminescence resonance energy transfer (CRET). When CEA exists, the specific combination of HRP-Apt and CEA can form HRP-Apt-CEA complex, which dissociates from GO. Then, the CL catalyzed by HRP-Apt-CEA complex can be detected without any CRET, and the content of CEA can be estimated by the CL intensity. It has been proved that the interference issue resulted from free HRP-Apt is solved well by mixing GO firstly with HRP-Apt, which blocks the free HRP-Apt's CL signal due to CL quenching effect of GO; and the interference resulted from GO to CL is also solved by CE, then the sensitivity and accuracy can be greatly improved. Results also showed that the CL intensity had a linear relationship with the concentration of CEA in the range from 0.0654 to 6.54 ng/mL, and the limit of detection was approximately 4.8 pg/mL (S/N = 3). This proposed method with high specificity offers a new way for separation and determination of biomolecule, and has good potential in application of biochemistry and bioanalysis.
基于适体/氧化石墨烯（Apt/GO）的毛细管电泳-化学发光（CE-CL）检测系统，提出了一种新的癌胚抗原（CEA）测定策略。首先，与辣根过氧化物酶（HRP）偶联的 CEA 适体与 GO 混合，由于 HRP-Apt 在 GO 上的堆积导致化学发光共振能量转移（CRET），CL 会被猝灭。当 CEA 存在时，HRP-Apt 与 CEA 的特异性结合可以形成 HRP-Apt-CEA 复合物，从而从 GO 上解离。然后，可以检测由 HRP-Apt-CEA 复合物催化的 CL，而没有任何 CRET，并且可以通过 CL 强度来估计 CEA 的含量。已经证明，通过首先将 GO 与 HRP-Apt 混合，可以很好地解决由于自由 HRP-Apt 引起的干扰问题，由于 GO 的 CL 猝灭效应，阻止了自由 HRP-Apt 的 CL 信号；并且通过 CE 还解决了 GO 对 CL 的干扰问题，从而大大提高了灵敏度和准确性。结果还表明，CL 强度与 CEA 浓度在 0.0654 至 6.54ng/mL 的范围内呈线性关系，检测限约为 4.8pg/mL（S/N = 3）。该方法具有高特异性，为生物分子的分离和测定提供了一种新方法，在生物化学和生物分析中有很好的应用潜力。
