Multiplex Immunochips for High-Accuracy Detection of AFP-L3% Based on Surface-Enhanced Raman Scattering: Implications for Early Liver Cancer Diagnosis
Multiplex Immunochips for High-Accuracy Detection of AFP-L3% Based on Surface-Enhanced Raman Scattering: Implications for Early Liver Cancer Diagnosis
复制标题
多重%20免疫芯片%20for%20高精度%20检测%20of%20AFP-L3%%20基于%20on%20表面增强%20拉曼%20散射:%20影响%20for%20早期%20肝脏%20癌症%20诊断
DOI：
10.1021/acs.analchem.7b01349
复制
发表时间：
2017-09-05
影响因子：
7.4
通讯作者：
He, Chengyan
中科院分区：
文献类型：
--
作者：
Ma, Hao;Sun, Xiaoying;He, Chengyan
甲胎蛋白（AFP）是一种重要的肿瘤生物标志物。特别是，AFP-L3的过表达与肝细胞癌（HCC）相关。因此，一些医院开始采用AFP-L3占总AFP水平的比值（AFP-L3%）作为诊断HCC的新证据，因为它具有较高的诊断准确性。然而，目前检测AFP和AFP-L3的方法耗时，需要多个样本，并且缺乏灵敏度和特异性。在此，我们提出了一个新的概念，肝癌的早期诊断的基础上结合拉曼频移和强度变化，并开发了表面增强拉曼散射（Sers）的免疫芯片通过AFP-L3%。在研究的第一步中，4-巯基苯甲酸（MBA）的频移用于定量测定总AFP的基于AFP和抗AFP的相互作用的MBA修饰的银芯片。然后将5，5-二硫代双（琥珀酰亚胺基-2-硝基苯甲酸酯）（DSNB）修饰的免疫金与AFP-L3抗体结合用于芯片上的夹心免疫反应。结果表明，DSNB的典型拉曼谱带强度与AFP-L3浓度呈指数线性关系。因此，AFP-L3%可以根据AFP-L3和总AFP的浓度来计算。该方法的最大优点是结合了AFP-L3%和Sers频移，具有良好的重现性和高准确性，大大简化了AFP-L3%的常规检测过程。应用所提出的方法与肝癌患者的血清显示其在早期肝癌诊断的巨大潜力。
alpha-Fetoprotein (AFP) is an important tumor biomarker. In particular, the overexpression of AFP-L3 is associated with hepatocellular carcinoma (HCC). Accordingly, several hospitals have begun to employ the ratio of AFP-L3 to the total AFP level (AFP-L3%) as new diagnostic evidence for HCC owing to its high diagnostic accuracy. However, current methods of detection for AFP and AFP-L3 are time-consuming, require multiple samples, and lack in sensitivity and specificity. Herein, we, present a novel concept for the early diagnosis of HCC based on the combination of Raman frequency shift and intensity change, and developed surface-enhanced Raman scattering (SERS)-based immunochips via AFP-L3%. In the first step of the study, the frequency shift of 4-mercaptobenzoic acid (MBA) was applied for the quantitative determination of total AFP based on the AFP and anti-AFP interaction on MBA-modified silver chips. 5,5-Dithiobis(succinimidy1-2-nitrobenzoate) (DSNB)-modified immunogold was then incorporated with AFP-L3 antibodies for sandwich immunoreaction on the chips. As a result, we found that a typical Raman band intensity of DSNB presented an exponential linear relationship with the concentration of AFP-L3. Thus, the AFP-L3% can be calculated according to the concentrations of AFP-L3 and total AFP. The most important advantage of the proposed method is the combination of AFP-L3% and frequency shifts Of SERS, which exhibits excellent reproducibility and high accuracy, and significantly simplifies the conventional detection procedure of AFP-L3%. Application of the proposed method with the serum of patients with HCC demonstrated its great potential in early liver cancer diagnosis.