Hu Weihua, Chen Hongming, Shi Zhuanzhuan, Yu Ling
Institute for Clean Energy and Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, China; Chongqing Key Laboratory for Advanced Materials and Technologies of Clean Energies, Southwest University, Chongqing 400715, China; Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), Wuhan University, Wuhan 430072, China.
Institute for Clean Energy and Advanced Materials, Faculty of Materials and Energy, Southwest University, Chongqing 400715, China; Chongqing Key Laboratory for Advanced Materials and Technologies of Clean Energies, Southwest University, Chongqing 400715, China.
Anal Biochem. 2014 May 15;453:16-21. doi: 10.1016/j.ab.2014.02.022. Epub 2014 Mar 4.
Surface plasmon resonance imaging (SPRi) is an intriguing technique for immunoassay with the inherent advantages of being high throughput, real time, and label free, but its sensitivity needs essential improvement for practical applications. Here, we report a dual signal amplification strategy using functional gold nanoparticles (AuNPs) followed by on-chip atom transfer radical polymerization (ATRP) for sensitive SPRi immunoassay of tumor biomarker in human serum. The AuNPs are grafted with an initiator of ATRP as well as a recognition antibody, where the antibody directs the specific binding of functional AuNPs onto the SPRi sensing surface to form immunocomplexes for first signal amplification and the initiator allows for on-chip ATRP of 2-hydroxyethyl methacrylate (HEMA) from the AuNPs to further enhance the SPRi signal. High sensitivity and broad dynamic range are achieved with this dual signal amplification strategy for detection of a model tumor marker, α-fetoprotein (AFP), in 10% human serum.
表面等离子体共振成像(SPRi)是一种用于免疫分析的有趣技术,具有高通量、实时和无标记的固有优势,但其灵敏度在实际应用中仍需大幅提高。在此,我们报告了一种双信号放大策略,该策略使用功能化金纳米颗粒(AuNPs),随后进行芯片上的原子转移自由基聚合(ATRP),用于人血清中肿瘤生物标志物的灵敏SPRi免疫分析。AuNPs接枝有ATRP引发剂以及识别抗体,其中抗体引导功能化AuNPs特异性结合到SPRi传感表面以形成免疫复合物进行首次信号放大,引发剂则允许从AuNPs进行甲基丙烯酸2-羟乙酯(HEMA)的芯片上ATRP,以进一步增强SPRi信号。通过这种双信号放大策略,在10%人血清中检测模型肿瘤标志物甲胎蛋白(AFP)时实现了高灵敏度和宽动态范围。