Jensen Gary C, Yu Xin, Gong Joseph D, Munge Bernard, Bhirde Ashwin, Kim Sang N, Papadimitrakopoulos Fotios, Rusling James F
Department of Chemistry, University of Connecticut, Storrs, CT 06269, USA.
J Nanosci Nanotechnol. 2009 Jan;9(1):249-55. doi: 10.1166/jnn.2009.j016.
Characterization studies of a multi-enzyme-antibody-carbon nanotube bioconjugate designed for the amplification of electrochemical immunosensing are described. Secondary antibodies for prostate specific antigen (PSA) were covalently linked to highly carboxylated multi-walled carbon nanotube (CNT) along with multiple horseradish peroxidase (HRP) enzyme labels. These bioconjugates provide ultra-sensitive amperometric detection of PSA on a single-wall carbon nanotube forest sandwich immunosensor platform. A single layer of HRP on the surface of the CNT was suggested by images from atomic force microscopy (AFM) and transmission electron microscopy (TEM). HRP on the bioconjugate surface was visualized by confocal microscopy using in-situ HRP-catalyzed polymerization yielding a fluorescent product, and HRP activity was estimated in a conventional assay. Binding of quantum-dot labeled PSA to antibodies on the bioconjugate was used for visualization by TEM. Combining TEM and enzyme activity results gave estimates of approximately 82 HRPs and 30 +/- 15 secondary antibodies per 100 nm of antibody-HRP-CNT bioconjugate.
本文描述了一种用于电化学免疫传感放大的多酶-抗体-碳纳米管生物共轭物的表征研究。前列腺特异性抗原(PSA)的二抗与多个辣根过氧化物酶(HRP)酶标记物一起共价连接到高度羧化的多壁碳纳米管(CNT)上。这些生物共轭物在单壁碳纳米管森林夹心免疫传感器平台上提供了对PSA的超灵敏安培检测。原子力显微镜(AFM)和透射电子显微镜(TEM)图像表明,CNT表面有单层HRP。通过共聚焦显微镜使用原位HRP催化聚合产生荧光产物来观察生物共轭物表面的HRP,并在传统检测中估计HRP活性。量子点标记的PSA与生物共轭物上抗体的结合用于通过TEM进行观察。结合TEM和酶活性结果估计,每100 nm的抗体-HRP-CNT生物共轭物中约有82个HRP和30±15个二抗。