【摘要】 目的:建立时间分辨荧光免疫层析法检测JWA蛋白的体系,并对该法进行评价。方法:运用EDC（1-（3-二甲氨基丙基）-3-乙基碳二亚胺盐酸盐）-NHS（N-羟基琥珀酰亚胺）活化时间分辨荧光微球,活化的时间分辨荧光微球按100μg Ab∶1 mg微球标记JWA单克隆抗体7C3;将荧光标记的抗体运用于免疫层析检测。结果:时间分辨荧光微球成功标记JWA单克隆抗体,荧光标记的7C3可以准确识别JWA多肽;测量范围可从20 ng·mL^-1至1500 ng·mL^-1,线性范围可达100 ng·mL^-1～1000 ng·mL^-1,回收率在95%以上,高、中、低浓度的重复性检测离散度（CV）均小于10%,加速稳定性亦较好,同时可以检测出胃癌细胞中的JWA蛋白。结论:本研究建立的时间分辨免疫荧光层析法测定JWA蛋白,各项分析性能良好,且便捷准确、快速经济,为检测JWA蛋白提供了一种高效的新途径。更多还原

【Abstract】 Objective: Using time-resolving fluorescent microspheres to label the JWA monoclonal antibodies and to establish the immunofluorescence chromatography strip. Methods: The EDC-NHS activated time-resolving fluorescent microspheres were used to label JWA monoclonal antibody 7 C3 with 100 μg Ab: 1 mg beads, and the fluorescently labeled antibodies were used for immunochromatographic detection. Results: The time-resolving fluorescent microspheres labeled JWA monoclonal antibody 7 C3 could accurately recognize JWA polypeptides, with a measuring range of 20-1500 ng·mL^-1. The linear response range of the method was 100-1000 ng·mL^-1. The values for coefficient of variation at high, medium and low concentrations were all less than 10% with average recovery rates over 95%. The JWA protein in gastric cancer cells can be detected simultaneously. Conclusion: The time-resolving immunofluorescence chromatographic method for assaying JWA protein in cells provided a convenient, rapid and economical option. It is a highly efficient new way for the detection of JWA protein.更多还原