Multiplexing determination of lung cancer biomarkers using electrochemical and surface-enhanced Raman spectroscopic techniques
Multiplexing determination of lung cancer biomarkers using electrochemical and surface-enhanced Raman spectroscopic techniques
复制标题
使用电化学和表面增强拉曼光谱技术多重测定肺癌生物标志物
DOI：
10.1039/c5nj00445d
复制
发表时间：
2015-01-01
影响因子：
3.3
通讯作者：
Qian, Weiping
中科院分区：
文献类型：
--
作者：
Lu, Wenbo;Wang, Ying;Qian, Weiping
研制了一种简单的电化学-表面增强拉曼光谱（Sers）复合免疫传感器，用于同时检测癌胚抗原（CEA）和细胞角蛋白19（CK-19）。通过π-π堆积作用，我们发现了一种拉曼染料（噻托溴铵和尼罗蓝A）修饰的树脂微球，它既有很强的Sers信号，又有电化学氧化还原特征峰。首次采用无模板法合成了氨基水杨酸树脂微球。采用还原法制备了Au纳米粒子（AuNP）包覆的AAR微球（AuNPs/AAR）。夹心结构包含具有第一抗体的拉曼染料标记的AuNPs/AAR、固定在电极修饰的壳聚糖稳定的AuNPs上的第二抗体和靶抗原。因此，在靶抗原的存在下，拉曼染料标记的AuNPs/AAR可以通过抗体-抗原-抗体相互作用结合到修饰电极表面。与噻托溴铵相比，耐尔蓝A具有不同的拉曼信号和电化学特征峰。因此，CEA和CK-19的同时免疫分析的基础上，开发了多标记的电化学和Sers免疫分析。该方法具有灵敏度高、选择性好、检测限低和长期稳定性好等特点。在信噪比为3时，CEA和CK-19的最低可检测浓度分别为0.01 ng mL−1和0.04 ng mL−1。该免疫传感器可用于人血清中CEA和CK-19的测定，结果良好。此外，电化学和Sers免疫传感器在肺癌诊断和治疗的临床研究领域具有潜在的应用前景。
A simple multiplexed electrochemical and surface-enhanced Raman spectroscopic (SERS) immunosensor is developed for simultaneous detection of carcinoembryonic antigen (CEA) and cytokeratin-19 (CK-19). We find a kind of Raman dye (thionine and Nile blue A)-decorated resin microspheres by π–π stacking interaction, which show both strong SERS signals and electrochemical redox characteristic peaks. Aminosalicylic acid-based resin (AAR) microspheres are synthesized by a template-free method for the first time. The Au nanoparticle (AuNP) coated AAR microspheres (AuNPs/AAR) are prepared by a convenient reduction approach. A sandwich structure contains Raman dye-labeled AuNPs/AAR with the first antibody, the second antibody immobilized on the electrode modified chitosan stabilized AuNPs, and target antigens. Thus, in the presence of the target antigens, the Raman dye-labeled AuNPs/AAR could be bonded to the modified electrode surface by antibody–antigen–antibody interactions. Compared with thionine, Nile blue A has different Raman signals and electrochemical characteristic peaks. As a result, a simultaneous immunoassay for CEA and CK-19 based on multiple labels is developed by using electrochemical and SERS immunoassays. The prepared immunoassay for detection of CEA and CK-19 shows high sensitivity, selectivity, low detection limit and long term stability. The lowest detectable concentration is 0.01 ng mL−1 and 0.04 ng mL−1 for CEA and CK-19 at a signal to noise rate of 3, respectively. The proposed immunosensor can be applied to determine CEA and CK-19 in human blood serum with favorable results. In addition, the electrochemical and SERS immunosensor has potential application in the diagnosis and treatment of lung cancer in the field of clinical research.