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富勒醇标记麦胚凝集素亲和吸附固体基质室温燐光法测定碱性磷酸酶

Triticum Vulgare Lectin Labeled with Fullerenol and its Determination of Trace Alkaline Phosphatase by Solid Substrate-room Temperature Phosphorimetry
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摘要 基于富勒醇在硝酸纤维素膜上能发射强而稳定的固体基质室温燐光信号,十二烷基苯磺酸钠能修饰富勒醇而发射更强的固体基质室温燐光,由此标记麦胚凝集素的产物,不仅能与碱性磷酸酶发生定量的特异性亲和吸附反应,而且亲和吸附反应产物能发射强而稳定的固体基质室温燐光信号,据此建立了十二烷基苯磺酸钠修饰富勒醇-麦胚凝集素-碱性磷酸酶-亲和吸附固体基质室温燐光测定碱性磷酸酶的新方法.方法的线性范围为0.040—45.00fg spot-1,工作曲线的回归亲程为△IP=32.68+5.514mALP fg spot-1,r=0.9994,检出限为0.011fg spot-1.本方法灵敏、准确、精密度好,用于人血清中碱性磷酸酶的测定与碱性磷酸酶试剂盒的检测结果相吻合. Fullerenol shows strong and stable room temperature phosphorescence (RTP) signal on the surface of nitrocelhdose membrane (NCM). When modified by dodecylbenzenesulfonic acid sodium salt (DBS), fullerenol emits a stronger signal, it was also found that quantitative specific affinity-adsorption reaction could be carried out between Triticum vulgare lectin (WGA) labeled with DBS-F and alkaline phosphatase (ALP) on the surface of NCM, and the product obtained (WGA-ALP-WGA-F-DBS) emits a strong and stable RTP signal. Furthermore, the content of ALP was proportional to the AlP value. Based on the facts above, a new method for the determination of trace amounts of ALP by affinity-adsorption solid-substrate room-temperature phosphorimetry (AA-SS-RTP) was established, using fallerenol modified with DBS to label WGA. The detection limit was 0.011 fg spot^-1. This method with high sensitivity, accuracy and precision, has been successfully applied to the determination of the content of ALP in human serum survey, and the results are tallied with those using alkaline phosphatasc kits.
作者 杨天隆
出处 《漳州师范学院学报(自然科学版)》 2009年第2期87-91,共5页 Journal of ZhangZhou Teachers College(Natural Science)
基金 福建省教育厅2007年第一批B类科技项目(JB07143)
关键词 碱性磷酸酶 富勒醇 标记 亲和吸附-固体基质室温燐光法 alkaline phosphatase fullerenol label room temperature phosphorescence method
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