Development of an enzyme linked immunosorbant assay for the detection of Cry2Ab Protein in transgenic plants.

GM crops Pub Date : 2011-04-01 DOI:10.4161/gmcr.2.2.16191
Suchitra Kamle, Abhishek Ojha, Arvind Kumar
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引用次数: 21

Abstract

Immunoassays' a sensitive technique has a wide application for detection of antigen, especially for the detection of growing genetically modified crops, which are underwent field trial. Here, we have developed for the quantitative detection of Cry2Ab protein expressed in GM crops. A Cry2Ab-rabbit-IgG act as capture antibody and Cry2Ab mouse monoclonal antibody behave as a detecting antibody, which is employed in sandwich ELISA. A 2% polyvinylpyrrollidone and 1% dithiothritol was utilized in protein extraction from cotton seed to avert interfering agents. The developed assay was validated with GM cotton samples, limit of detection was found 1 pg/ml and range to a limit of quantification 16 pg/ml. The developed immunoassay does not show any cross reactivity with other non target GM proteins like Cry1Ab, Cry1Ac, EPSPS, Vip.

转基因植物中Cry2Ab蛋白酶联免疫吸附试验的建立。
免疫测定法是一种灵敏的抗原检测技术,在检测正在生长的转基因作物中有着广泛的应用。在此,我们开发了转基因作物中表达的Cry2Ab蛋白的定量检测方法。Cry2Ab兔igg作为捕获抗体,小鼠Cry2Ab单克隆抗体作为检测抗体,采用夹心ELISA法。以2%聚乙烯吡咯烷酮和1%二硫代四醇为溶剂,对棉籽中蛋白质的提取进行了研究。用转基因棉花样品验证了该方法,检测限为1 pg/ml,定量限为16 pg/ml。该方法与Cry1Ab、Cry1Ac、EPSPS、Vip等非靶基因蛋白无交叉反应性。
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