血清中可溶性白介素-2受体胶体金免疫层析试纸检测方法研究  

Preparation of Colloidal Gold-Labeled Immunochromatographic Strips for Detection of Soluble Interleukin-2 Receptors in Real Samples

作　　者：徐凤琴 朱群艳 张婳[1] 王振新[1,2] XU Feng-Qin;ZHU Qun-Yan;ZHANG Hua;WANG Zhen-Xin(State Key Laboratory of Electroanalytical Chemistry,Changchun Institute of Applied Chemistry,Chinese Academy of Sciences,Changchun 130022,China;School of Applied Chemical Engineering,University of Science and Technology of China,Hefei 230026,China)

机构地区：[1]中国科学院长春应用化学研究所电分析化学国家重点实验室,长春130022 [2]中国科学技术大学应用化学与工程学院,合肥230026

出　　处：《分析化学》2021年第6期973-981,共9页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金项目(No.61901438)资助。

摘　　要：快速、灵敏地检测血清中的可溶性白介素-2受体(sIL-2R)对相关疾病的临床诊断至关重要。本研究建立了一种基于胶体金免疫层析试纸条的快速、定量检测实际样品中sIL-2R的方法。在硝酸纤维膜上分别包被羊抗兔二抗(2ndAb)和兔抗鼠sIL-2R单克隆抗体(cAb)作为质控线(C线)和检测线(T线);以兔抗鼠sIL-2R单克隆抗体(dAb)修饰的金纳米粒子为标记物,采用双抗体夹心法制备用于sIL-2R检测的免疫层析试纸条。合成了3种不同形貌和粒径的金纳米粒子,考察了其对胶体金标记免疫层析试纸条检测性能的影响。实验结果表明,胶体金标记免疫层析试纸条对sIL-2R的检测性能与金纳米粒子的形貌和粒径密切相关;其中,金纳米棒(Gold nanorod,GNR)标记的免疫层析试纸条具有较宽的线性范围(1~6250 ng/mL)和较低的检出限(1.0 ng/mL)。采用GNR免疫层析试纸条成功实现了荷三阴性乳腺肿瘤(MDA-MB-231)BALB/c裸鼠血清中sIL-2R浓度的检测,结果表明,sIL-2R浓度与肿瘤进程呈正相关。Rapid and sensitive detection of soluble interleukin-2 receptor(sIL-2R)in serum is believed to be of vital importance in clinical diagnosis.In this study,a colloidal gold-labeled immunochromatographic strip was developed for rapid and quantitative detection of sIL-2R in real sample.The goat anti-rabbit secondary antibody(2ndAb)and rabbit anti-mouse sIL-2R monoclonal antibody(cAb)were sprayed onto the nitrocellulose membrane as control line(C line)and test line(T line),respectively.The rabbit anti-mouse sIL-2R monoclonal antibody(dAb)modified gold nanoparticles were used as markers to fabricate the immunochromatographic strip for detection of sIL-2R by a sandwich method.Three gold nanoparticles with different sizes and morphologies were synthesized,and their effects on the detection performance of colloidal gold-labeled immunochromatographic strip were investigated.The experimental results showed that the detection performance of colloidal gold-labeled immunochromatographic strip for sIL-2R was closely related to the morphology and particle size of gold nanoparticles.Among them,gold nanorod(GNR)-based immunochromatographic strip had the widest linear range(1-6250 ng/mL)and the lowest detection limit(1.0 ng/mL).In addition,the concentrations of sIL-2R in the sera of triple-negative breast tumors(MDA-MB-231)-bearing BALB/c nude mice were successfully detected using GNR-based immunochromatographic strips,and it was found that the concentration of sIL-2R was positively correlated with tumor progression.

关 键 词：金纳米粒子 免疫层析 可溶性白介素-2受体 恶性肿瘤 

分 类 号：R446.61[医药卫生—诊断学]