用于检测毒莠定除草剂的绿色荧光蛋白标记重组氟抗体

Green fluorescent protein-labeled recombinant fluobody for detecting the picloram herbicide.

作者信息

Kim In Seon, Shim Jae Han, Suh Yong Tack, Yau Kerrm Y F, Hall J Christopher, Trevors Jack T, Lee Hung

机构信息

Department of Chemical and Materials Engineering, University of Alberta, Edmonton, Canada.

出版信息

Biosci Biotechnol Biochem. 2002 May;66(5):1148-51. doi: 10.1271/bbb.66.1148.

DOI:10.1271/bbb.66.1148

PMID:12092834

Abstract

A green fluorescent protein-labeled fluobody was designed to develop a simple immunoassay method for detecting picloram herbicide in an environmental sample. The gfp gene was successfully inserted into the pSJF2 vector harboring the picloram-specific antibody fragment to yield pSJF2GFP. Picloram spiking in an environmental river sample could be indirectly detected by observing the fluorescence intensity value of the gfp-fluobody, exhibiting specific sensitivity to free picloram with an IC50 value of 50 ppb. Using the gfp-fluobody immunoassay avoids the enzyme-substrate reaction for calorimetric detection that is required in an enzyme-linked immunosorbent assay (ELISA).

摘要

设计了一种绿色荧光蛋白标记的氟抗体，以开发一种用于检测环境样品中吡氯草酮除草剂的简单免疫分析方法。绿色荧光蛋白基因成功插入携带吡氯草酮特异性抗体片段的PSJF2载体中，得到PSJF2GFP。通过观察绿色荧光蛋白 - 氟抗体的荧光强度值，可以间接检测环境河流样品中添加的吡氯草酮，对游离吡氯草酮表现出特异性敏感性，半数抑制浓度值为50 ppb。使用绿色荧光蛋白 - 氟抗体免疫分析避免了酶联免疫吸附测定（ELISA）中用于比色检测所需的酶 - 底物反应。