基于磁珠支撑双酶催化质量增强策略的高灵敏度石英晶体微天平免疫传感器。
A highly sensitive quartz crystal microbalance immunosensor based on magnetic bead-supported bienzymes catalyzed mass enhancement strategy.
机构信息
Department of Chemistry, Chungnam National University, Daejeon 305-764, South Korea.
Department of Chemistry, Chungnam National University, Daejeon 305-764, South Korea; Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon 305-764, South Korea.
出版信息
Biosens Bioelectron. 2015 Apr 15;66:539-46. doi: 10.1016/j.bios.2014.12.007. Epub 2014 Dec 3.
A highly sensitive quartz crystal microbalance (QCM) immunosensor based on magnetic bead-supported bienzyme catalyzed mass enhanced strategy was developed for the detection of human immunoglobulin G (hIgG) protein. The high sensitive detection was achieved by increasing the deposited mass on the QCM crystal through the enhanced precipitation of 4-chloro-1-naphthol (CN) using higher amounts of horseradish peroxidase (HRP) and glucose oxidase (GOx) bienzymes attached on the magnetic beads (MB). The protein A (PA) and capture antibody (monoclonal anti-human IgG antibody produced in mouse, Ab1)-based QCM probe and the detection antibody (anti-human IgG antibody produced in goat, Ab2)-based MB/HRP/GOx bienzymatic bioconjugates were characterized using scanning electron microscope, transmission electron microscope, cyclic voltammetry, and electrochemical impedance spectroscopy techniques. Under the optimized experimental condition, the linear range and the detection limit of hIgG immunosensor were determined to be 5.0pg/mL-20.0ng/mL and 5.0±0.18pg/mL, respectively. The applicability of the present hIgG immunosensor was examined in hIgG spiked human serum samples and excellent recoveries of hIgG were obtained.
基于磁珠支撑双酶催化质量增强策略的高灵敏度石英晶体微天平（QCM）免疫传感器用于检测人免疫球蛋白 G（hIgG）蛋白。通过使用更多的辣根过氧化物酶（HRP）和葡萄糖氧化酶（GOx）双酶附着在磁珠上，增强 4-氯-1-萘酚（CN）的沉淀，从而在 QCM 晶体上增加沉积质量，实现了高灵敏度检测。基于蛋白 A（PA）和捕获抗体（鼠源单克隆抗人 IgG 抗体，Ab1）的 QCM 探针和基于检测抗体（山羊源抗人 IgG 抗体，Ab2）的磁珠/HRP/GOx 双酶生物缀合物用扫描电子显微镜、透射电子显微镜、循环伏安法和电化学阻抗谱技术进行了表征。在优化的实验条件下，确定 hIgG 免疫传感器的线性范围和检测限分别为 5.0pg/mL-20.0ng/mL 和 5.0±0.18pg/mL。本 hIgG 免疫传感器在 hIgG 加标人血清样品中的适用性进行了检验，获得了 hIgG 的优异回收率。
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