基于双重信号放大的电化学免疫法对前列腺特异性抗原的检测  

Detection of prostate specific antigen-based on dual signal amplified electrochemical immunoassay

作　　者：李雪 艾永玲 何群叶 杨功俊 王菁[1,2] LI Xue;AI Yongling;HE Qunye;YANG Gongjun;WANG Jing(Key Laboratory of Biomedical Functional Materials,Ministry of Education,China Pharmaceutical University,Nanjing 210009,China;Key Laboratory of Drug Quality Control and Pharmacovigilance,Ministry of Education,China Pharmaceutical University,Nanjing 210009,China)

机构地区：[1]中国药科大学医药生物功能材料重点实验室,南京210009 [2]中国药科大学药物质量与安全预警教育部重点实验室,南京210009

出　　处：《中国药科大学学报》2018年第4期433-440,共8页Journal of China Pharmaceutical University

基　　金：国家自然科学基金资助项目(No.21775165);江苏省自然科学基金资助项目(No.BK20161455);中国药科大学药学基地科研训练及科研能力提高项目资助(No.J1310032)~~

摘　　要：以谷胱甘肽修饰的CdTe量子点(GSH-CdTeQDs)与金纳米粒(Au NPs)形成复合材料(AuNPs@GSH-CdTe)为信号标记物,并在还原氧化石墨烯(rGO)和AuNPs双重信号放大的作用下,建立了一种高灵敏检测前列腺特异性抗原(PSA)的三明治免疫夹心式电化学方法。在rGO/AuNPs表面固定二抗蛋白(Ab_2),捕获目标物PSA和标记有一抗蛋白(Ab1)的AuNPs@GSH-CdTe信号物,形成"三明治"免疫夹心结构。经HNO_3溶解后,采用方波溶出伏安法(SWSV)测定酸解的Cd^(2+)的峰电流用于定量分析PSA。其中AuNPs较大的比表面积以及较好的生物相容能力,达到了成功装载抗体以及放大信号的效果,同时具有较大表面积的rGO起到了协同放大的作用。所构建的免疫分析方法实现了对肿瘤标志物PSA的检测,其线性范围为0.5~200ng/mL,检测限为5.0 pg/mL,并且该方法专属性、重复性以及稳定性好。此外,在实际样品的检测中,加标样回收率为98.20%~106.2%,结果准确度良好,为检测PSA提供了准确可靠且灵敏度高的新方法。A sandwiched electrochemical immunoassay based on the AuNPs@GSH-CdTe as a signal label,which formed by GSH-CdTe QDs and AuNPs,with dual signal amplified by reduced graphene oxide and AuNPs was proposed for the sensitive detection of prostate specific antigen(PSA).Through a sandwich immunoreaction,the target PSA and AuNPs@GSH-CdTe labeled Ab 1 were captured to rGO/AuNPs-Ab 2 surface.After the HNO 3-dissolution step,square wave stripping voltammetry(SWSV)analysis of the captured CdTe QDs was used to quantify the concentration of PSA.In this system,AuNPs possessedlarge specific surface and good biocompatibility,which could effectively expand the amount of antigen and GSH-CdTe QDs loading and signals amplifying,while rGO played a synergistic amplification role due to its large specific surface.The proposed method showed good linearity ranging from 0.5 to 200 ng/mL with the detection limits of 5.0 pg/mL.It also showed excellent selectivity,good reproducibility,satisfactory stability.In addition,the method was successfully applied to the determination of real samples.The result was satisfactory and the recovery could fall in 98.20%106.2%,which represented a novel approach for versatile detection of tumor markers.

关 键 词：方波溶出伏安法 GSH-CdTe量子点 金纳米粒 还原氧化石墨烯 双重信号 前列腺特异性抗原 

分 类 号：R917[医药卫生—药物分析学]