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基于介孔硅结构上掺杂金纳米粒子的 MCF-7 乳腺癌细胞裂解液和未经处理人血浆中肿瘤蛋白 CA 15.3 的超灵敏免疫分析

Ultrasensitive immunoassay of tumor protein CA 15.3 in MCF-7 breast cancer cell lysates and unprocessed human plasma using gold nanoparticles doped on the structure of mesoporous silica.

机构信息

Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.

Department of Biochemistry, Higher Education Institute of Rab-Rashid, Tabriz, Iran.

出版信息

Int J Biol Macromol. 2018 Dec;120(Pt B):2493-2508. doi: 10.1016/j.ijbiomac.2018.09.020. Epub 2018 Sep 5.

Abstract

Breast cancer is the most common threat in women worldwide. Increasing death rate of diagnosed cases is the main leading cause of designing specific immunoassay for tumor marker in breast cancer. Cancer antigen 15-3 (CA 15-3) is a tumor protein for many types of cancer, most notably breast cancer. Herein, we report a sandwich-type electrochemical immunosensor based on signal amplification strategy of multiple nanocomposites to detection of CA 15-3 biomarker. The proposed immunosensor fabricated by reduced graphene oxide (rGO), poly-dopamine (PDA) and amino functionalized mesoporous silica (MCM-41-NH) doped by gold nanoparticles composite on the glassy carbon electrode with a large surface area which was prepared a new platform to immobilization of primary antibodies and provide excellent conductivity. To further amplify the electrochemical signal, the trace tag on the foundation of gold nanoparticles (AuNPs) is coated with MCM-41-NH nanocomposite through thionine linking, which provides more amino groups to capture more horseradish peroxidase-labeled antibodies (HPR-Ab2) and enhances the conductivity. Under optimal conditions, the developed immunosensor exhibits excellent analytical performance for the determination of CA 15-3 with a wide linear range from 0.002 to 125 U/mL and a low limit of quantification of 0.002 U/mL. Furthermore, satisfactory results are obtained for the determination of CA 15-3 in human plasma samples, indicating the potential of the immunoassay to be applied in clinical analysis.

摘要

乳腺癌是全球女性最常见的威胁。诊断病例死亡率的上升是设计针对乳腺癌肿瘤标志物的特定免疫分析的主要原因。癌抗原 15-3(CA 15-3)是许多类型癌症的肿瘤蛋白,尤其是乳腺癌。在此,我们报告了一种基于信号放大策略的基于三明治型电化学免疫传感器的多重纳米复合材料用于检测 CA 15-3 生物标志物。所提出的免疫传感器由还原氧化石墨烯(rGO)、聚多巴胺(PDA)和氨基功能化介孔硅(MCM-41-NH)组成,掺杂纳米金复合在玻碳电极上,具有较大的表面积,用于固定化的主要抗体和提供优良的导电性。为了进一步放大电化学信号,在金纳米粒子(AuNPs)的基础上涂覆介孔硅纳米复合材料,通过硫堇连接,提供更多的氨基来捕获更多辣根过氧化物酶标记的抗体(HPR-Ab2)并增强导电性。在最佳条件下,所开发的免疫传感器对 CA 15-3 的测定具有出色的分析性能,线性范围从 0.002 到 125 U/mL,检测限低至 0.002 U/mL。此外,在人血浆样品中测定 CA 15-3 得到了令人满意的结果,表明该免疫分析具有应用于临床分析的潜力。

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