State Key Laboratory of Analytical Chemistry for Life Science, Department of Chemistry, Nanjing University, Nanjing 210093, People's Republic of China.
Anal Chem. 2012 Mar 6;84(5):2410-5. doi: 10.1021/ac203179g. Epub 2012 Feb 21.
A sensitive chemiluminescence (CL) imaging immunoassay method for detection of multiple tumor markers with high throughput, easy operation, and low cost was developed. The immunosensor array was prepared by covalently immobilizing capture antibodies on corresponding sensing sites on a silanized disposable glass chip. Gold nanoparticle-based bioconjugates with a high molar ratio of horseradish peroxidase (HRP) to detection antibodies were used for signal amplification. Under a sandwich immunoassay, the CL signals triggered by HRP captured on each sensing cell were collected by a charge-coupled device for simultaneous measurement of biomarkers and combination diagnosis of certain tumors. As a proof of concept, the immunosensor array was applied to detect α-fetoprotein, carcinoma antigen 125, carbohydrate antigen 153, and carcinoembryonic antigen and to screen patients with liver, breast, or ovarian cancers. This method showed wide linear ranges over 5 orders of magnitude and much lower detection limits than previously reported multiplexed immunoassays. The high throughput and acceptable stability, reproducibility, and accuracy showed good applicability of the proposed multiplex CL imaging immunoassay in clinical diagnosis.
开发了一种用于检测多种肿瘤标志物的高灵敏度、操作简单、成本低廉的化学发光（CL）成像免疫分析方法。将捕获抗体通过共价键固定在硅烷化的一次性玻璃芯片的相应传感点上，制备免疫传感器阵列。基于金纳米粒子的生物缀合物具有高摩尔比的辣根过氧化物酶（HRP）与检测抗体，用于信号放大。在三明治免疫分析中，通过 CCD 收集每个传感单元上捕获的 HRP 触发的 CL 信号，用于同时测量生物标志物和对某些肿瘤进行联合诊断。作为概念验证，免疫传感器阵列用于检测甲胎蛋白、癌抗原 125、糖链抗原 153 和癌胚抗原，并筛选肝癌、乳腺癌或卵巢癌患者。与先前报道的多重免疫分析相比，该方法在 5 个数量级范围内具有较宽的线性范围和更低的检测限。高通量以及可接受的稳定性、重现性和准确性表明，所提出的多重 CL 成像免疫分析在临床诊断中具有良好的适用性。
