Rational design and application of broad-spectrum antibodies for Bt Cry toxins determination
文献类型： 外文期刊
作者： Jin, Jiafeng 1 ; Chen, Wei 1 ; Xu, Chongxin 1 ; Pooe, Ofentse Jacob 4 ; Xie, Yajing 1 ; Shen, Cheng 1 ; Meng, Meng 1 ; Zhu, Qin 1 ; Zhang, Xiao 1 ; Liu, Xianjin 1 ; Liu, Yuan 1 ;
作者机构： 1.Jiangsu Acad Agr Sci, Inst Food Safety & Nutr, Jiangsu Key Lab Food Qual & Safety, State Key Lab Cultivat Base Minist Sci & Technol, Nanjing 210014, Peoples R China
2.Jiangsu Univ, Sch Food & Biol Engn, Zhenjiang 212013, Peoples R China
3.Nanjing Agr Univ, Coll Plant Protect, Nanjing 210023, Peoples R China
4.Univ KwaZulu Natal, Sch Life Sci, ZA-4000 Durban, South Africa
5.Jiangsu Univ, Sch Life Sci, Zhenjiang 212013, Peoples R China
关键词： Bt Cry toxins; Polyclonal antibodies; Monoclonal antibody; ELISA; Genetically modified crops
期刊名称：ANALYTICAL BIOCHEMISTRY （ 影响因子：2.6； 五年影响因子：2.7 ）
ISSN： 0003-2697
年卷期： 2024 年 693 卷
页码：
收录情况： SCI
摘要： Using the amino acid sequences and analysis of selected known structures of Bt Cry toxins, Cry1Ab, Cry1Ac, Cry1Ah, Cry1B, Cry1C and Cry1F we specifically designed immunogens. After antibodies selection, broadspectrum polyclonal antibodies (pAbs) and monoclonal antibody (namely 1A0-mAb) were obtained from rabbit and mouse, respectively. The produced pAbs displayed broad spectrum activity by recognizing Cry1 toxin, Cry2Aa, Cry2Ab and Cry3Aa with half maximal inhibitory concentration (IC 50 ) values of 0.12 -9.86 mu g/mL. Similarly, 1A0-mAb showed broad spectrum activity, recognizing all of the above Cry protein (IC 50 values of 4.66 -20.46 mu g/mL) with the exception of Cry2Aa. Using optimizations studies, 1A10-mAb was used as a capture antibody and pAbs as detection antibody. Double antibody sandwich enzyme -linked immunosorbent assays (DAS-ELISAs) were established for Cry1 toxin, Cry2Ab and Cry3Aa with the limit of detection (LOD) values of 2.36 -36.37 ng/mL, respectively. The present DAS-ELISAs had good accuracy and precisions for the determination of Cry toxin spiked tap water, corn, rice, soybeans and soil samples. In conclusion, the present study has successfully obtained broad-spectrum pAbs and mAb. Furthermore, the generated pAbs- and mAb-based DASELISAs protocol can potentially be used for the broad-spectrum monitoring of eight common subtypes of Bt Cry toxins residues in food and environmental samples.
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