Kim Namsoo, Kim Dong-Kyung, Cho Yong-Jin, Moon Dae-Kyung, Kim Woo-Yeon
Food Nano-Biotechnology Research Center, Korea Food Research Institute, Songnam-Si, Kyonggi-Do, Republic of Korea.
Biosens Bioelectron. 2008 Nov 15;24(3):391-6. doi: 10.1016/j.bios.2008.04.013. Epub 2008 Apr 26.
A label-free carp vitellogenin sensor has a strong potential for on-site monitoring on the possible contamination of edible fish with endocrine disruptors as a sum parameter in an inland carp farm. In this study, we performed a sensitive detection for carp vitellogenin with a direct-binding optical waveguide lightmode spectroscopy-based immunosensor. Carp vitellogenin bound over the sensor surface quite specifically, judging from the sensor responses according to stepwise antibody immobilization. This was also supported by a negligible sensor response found at bovine serum albumin immobilization. When plotted in double-logarithmic scale for carp vitellogenin concentrations of 0.00675-67.5 nM, a linear relationship was found between analyte concentration and sensor response, together with the limit of detection of 0.00675 nM. The reusability of the immunosensor after the regeneration with 10mM HCl was reasonably good, as presumed from the coefficient of variability of 6.02% for nine repetitive measurements. The model sample prepared by spiking a purified carp vitellogenin into a 10-fold diluted vitellogenin-free carp serum in 9.45 nM showed the response ratio of 96.70% against 9.45 nM of the purified carp vitellogenin. When a female and male carp sera induced with 17beta-estradiol injection were analyzed, biomarker induction was even identifiable at 2000-fold serum dilution.
一种无标记的鲤鱼卵黄蛋白原传感器在内陆鲤鱼养殖场中作为总和参数对食用鱼可能受到内分泌干扰物污染进行现场监测具有很大潜力。在本研究中,我们使用基于直接结合光波导光模式光谱的免疫传感器对鲤鱼卵黄蛋白原进行了灵敏检测。从逐步固定抗体后的传感器响应判断,鲤鱼卵黄蛋白原在传感器表面特异性结合。在固定牛血清白蛋白时传感器响应可忽略不计也支持了这一点。当以双对数尺度绘制鲤鱼卵黄蛋白原浓度为0.00675 - 67.5 nM的曲线时,发现分析物浓度与传感器响应之间存在线性关系,检测限为0.00675 nM。用10mM HCl再生后免疫传感器的可重复使用性相当好,从九次重复测量的变异系数6.02%可以推断。通过将纯化的鲤鱼卵黄蛋白原加入到9.45 nM的10倍稀释的无卵黄蛋白原鲤鱼血清中制备的模型样品,对9.45 nM纯化鲤鱼卵黄蛋白原的响应率为96.70%。当分析用17β - 雌二醇注射诱导的雌性和雄性鲤鱼血清时,即使在血清稀释2000倍时也能识别生物标志物诱导情况。