基于适配体-抗体杂交用于干扰素-γ的SERS高灵敏检测  

Aptamer-Antibody hybridization-based SERS sensitive detection of IFN-γ

作　　者：金伽利 陈燕[2] 杨丹婷 JIN Jiali;CHEN Yan;YANG Danting(School of Public Health,School of Medicine,Ningbo University,Ningbo,Zhejiang 315211,P.R.China;The Affiliated Lihuili Hospital,Ningbo University,Ningbo,315000)

机构地区：[1]宁波大学医学院公共卫生医学院,宁波315211 [2]宁波大学附属李惠利医院,宁波315000

出　　处：《光散射学报》2023年第4期383-390,共8页The Journal of Light Scattering

基　　金：浙江省自然科学基金(LY23H260002);国家自然科学基金(82073514);浙江省省属高校基本科研业务费专项资金(SJLY2021009);宁波大学王宽诚基金资助项目。

摘　　要：干扰素-γ是一种重要的细胞因子,可作为临床上疾病预防、诊断和预后检测的生物标记物。为了实现对其的灵敏检测,本文基于抗体和适配体的杂交结构,开发了一种适配体传感器,其中涉及了两种探针。首先,将4-硫基苯甲酸嵌入金银合金纳米颗粒后将其与干扰素-γ适配体偶联作为检测探针。其次,将干扰素-γ抗体修饰的磁珠作为捕获探针。当干扰素-γ存在时,干扰素-γ蛋白可同时被适配体和抗体捕获,形成稳定的三明治结构,获得较强的拉曼信号;反之,当干扰素-γ不存在时,则获得较弱的拉曼信号。我们对相关的重要参数如4-硫基苯甲酸的体积、适配体的浓度、以及抗体的体积进行了优化。实验结果表明,该方法具有高灵敏度和较好的重复性,其检测限为0.25 pg/mL,在血清中的回收率高达98.7%~108%。因此这一抗体/适配体杂交传感器能实现在复杂血清环境中对干扰素-γ的快速、准确的检测,以满足临床需要。Interferon-γ(IFN-γ)is an important cytokine,which can be used as a biomarker for disease prevention,diagnosis,and prognosis detection in clinics.To achieve highly sensitive detection of IFN-γ,an aptasensor based on hybrid structure of antibodies and aptamer was developed,which involves two probes.Firstly,4-Mercaptobenzoic acid(4-MBA)was embedded into Au-Ag Alloy nanoparticles,and then coupled with the aptamer as detection probes.Secondly,magnetic beads modified by antibodies were used as capture probes.When IFN-γis present,IFN-γcan be captured by both aptamers and antibodies,forming a stable sandwich structure,and obtaining strong Raman signals.On the contrary,when there is not IFN-γ,weak Raman signals are obtained.We optimized the relevant parameters such as volume of 4-MBA and antibody,concentration of aptamer.The experimental results show that this method has high sensitivity and repeatability,with a limit of detection(LOD)of 0.25 pg/mL in PBS.Due to the high recovery rate of this method in serum ranging from 98.7% to 108%,we believe that this aptasensor can achieve the fast and accurate detection of interferon-γin complex serum environments to meet clinical needs.

关 键 词：表面增强拉曼散射 适配体 干扰素-Γ 三明治杂交 金银合金纳米颗粒 

分 类 号：O657.37[理学—分析化学]