Development of Monoclonal Antibodies Recognizing Linear Epitope: Illustration by Three Bacillus thuringiensis Crystal Proteins of Genetically Modified Cotton, Maize, and Tobacco
文献类型: 外文期刊
第一作者: Cao, Zhen
作者: Cao, Zhen;Zhang, Wei;Ning, Xiangxue;Wang, Baomin;Cao, Zhen;Li, Qing X.;Liu, Yunjun
作者机构:
关键词: CrylAc;Cryllal;Collie;GMO;Western blot;linear epitope;antibody
期刊名称:JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY ( 影响因子:5.279; 五年影响因子:5.269 )
ISSN: 0021-8561
年卷期: 2017 年 65 卷 46 期
页码:
收录情况: SCI
摘要: Bacillus thuringiensis CrylAc, CrylIal, and Crylle are S-endotoxin insecticidal proteins widely implemented in genetically modified organisms (GMO), such as cotton, maize, and potato. Western blot assay integrates electrophoresis separation power and antibody high specificity for monitoring specific exogenous proteins expressed in GMO. Procedures for evoking monoclonal antibody (mAb) for Western blot were poorly documented. In the present study, CrylAc partially denatured at 100 degrees C for 5 min was used as an immunogen to develop mAbs selectively recognizing a linear epitope of CrylAc for Western blot. mAb 5E9C6 and 3E6E2 selected with sandwich ELISA strongly recognized the heat semidenatured CrylAc. Particularly, 3E6E2 recognized both E. coli and cotton seed expressed CrylAc in Western blot. Such strategy of using partially denatured proteins as immunogens and using sandwich ELISA for mAb screening was also successfully demonstrated with production of mAbs against Crylle for Western blot assay in maize.
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