Center for Research at the Bio/Nano Interface, Department of Chemistry and Shands Cancer Center, UF Genetics Institute and McKnight Brain Institute, University of Florida, Gainesville, FL 32611, USA.
Anal Biochem. 2011 Mar 1;410(1):124-32. doi: 10.1016/j.ab.2010.11.005. Epub 2010 Nov 13.
Biosensors based on antibody recognition have a wide range of monitoring applications that apply to clinical, environmental, homeland security, and food problems. In an effort to improve the limit of detection of the Naval Research Laboratory (NRL) Array Biosensor, magnetic nanoparticles (MNPs) were designed and tested using a fluorescence-based array biosensor. The MNPs were coated with the fluorescently labeled protein, AlexaFluor647-chicken IgG (Alexa647-chick IgG). Antibody-labeled MNPs (Alexa647-chick-MNPs) were used to preconcentrate the target via magnetic separation and as the tracer to demonstrate binding to slides modified with anti-chicken IgG as a capture agent. A full optimization study of the antibody-modified MNPs and their use in the biosensor was performed. This investigation looked at the Alexa647-chick-MNP composition, MNP surface modifications, target preconcentration conditions, and the effect that magnetic extraction has on the Alexa647-chick-MNP binding with the array surface. The results demonstrate the impact of magnetic extraction using the MNPs labeled with fluorescent proteins both for target preconcentration and for subsequent integration into immunoassays performed under flow conditions for enhanced signal generation.
基于抗体识别的生物传感器具有广泛的监测应用，适用于临床、环境、国土安全和食品安全问题。为了提高海军研究实验室（NRL）阵列生物传感器的检测极限，设计并测试了磁性纳米粒子（MNPs），使用基于荧光的阵列生物传感器。MNPs 用荧光标记的蛋白质 AlexaFluor647-鸡 IgG（Alexa647-chick IgG）进行了涂层处理。抗体标记的 MNPs（Alexa647-chick-MNPs）用于通过磁分离预浓缩目标，并作为示踪剂来证明与作为捕获剂的抗鸡 IgG 修饰的载玻片结合。对抗体修饰的 MNPs 及其在生物传感器中的应用进行了全面的优化研究。这项研究考察了 Alexa647-chick-MNP 的组成、MNP 表面修饰、目标预浓缩条件，以及磁提取对 Alexa647-chick-MNP 与阵列表面结合的影响。结果表明，使用标记有荧光蛋白的 MNPs 进行磁提取，无论是用于目标预浓缩还是用于随后在流动条件下进行免疫分析以增强信号产生，都具有重要意义。
