Korea Research Institute of Bioscience and Biotechnology, Yuseong, Daejeon 305-600, Republic of Korea.
Anal Biochem. 2011 Jan 15;408(2):206-11. doi: 10.1016/j.ab.2010.09.026. Epub 2010 Sep 22.
We describe a method of amplifying the biosensing signal in surface plasmon resonance (SPR)-based immunoassays using an antibody-carbon nanotube (CNT) conjugate. As a model system, human erythropoietin (EPO) and human granulocyte macrophage colony-stimulating factor (GM-CSF) were detected by sandwich-type immunoassays using an SPR biosensor. For the amplification of the SPR signal, the CNT was conjugated with a polyclonal antibody, and then the conjugates were reacted with antibodies coupled with the target proteins. This amplification strategy increases the dynamic range of the immunoassays and enhances the detection sensitivity. The SPR immunoassays, combined with the CNT-assisted signal amplification method, provided a wide dynamic range over four orders of magnitude for both EPO and GM-CSF (0.1-1,000 ng/ml). The CNT amplification method is expected to realize the detection of picogram levels and a wide dynamic detection range of multiple proteins, enabling it to offer a robust analysis tool for the development of biopharmaceutical production.
我们描述了一种使用抗体-碳纳米管(CNT)缀合物在基于表面等离子体共振(SPR)的免疫分析中放大生物传感信号的方法。作为模型系统,使用 SPR 生物传感器通过夹心型免疫分析检测人红细胞生成素(EPO)和人粒细胞-巨噬细胞集落刺激因子(GM-CSF)。为了放大 SPR 信号,将 CNT 与多克隆抗体缀合,然后将缀合物与与靶蛋白偶联的抗体反应。这种放大策略增加了免疫分析的动态范围,并提高了检测灵敏度。SPR 免疫分析与 CNT 辅助的信号放大方法相结合,为 EPO 和 GM-CSF(0.1-1,000ng/ml)提供了四个数量级的宽动态范围。CNT 放大方法有望实现皮克级水平的检测和多种蛋白质的宽动态检测范围,为生物制药生产的发展提供了强大的分析工具。