Development of a monoclonal antibody-based competitive ELISA for detection of β-conglycinin, an allergen from soybean
Development of a monoclonal antibody-based competitive ELISA for detection of β-conglycinin, an allergen from soybean
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DOI：
10.1016/j.foodchem.2007.05.040
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发表时间：
2008
期刊：
影响因子：
8.8
通讯作者：
J. You;Defa Li;S. Qiao;Zirui Wang;Pingli He;D. Ou;B. Dong
中科院分区：
文献类型：
--
作者：
J. You;Defa Li;S. Qiao;Zirui Wang;Pingli He;D. Ou;B. Dong
大豆β-甘氨酸是儿童和幼小动物的主要食物过敏之一。为了检测大豆及大豆制品中具有免疫反应性的β-conglycin，以合成的与β-conglycin表位序列对应的肽段为免疫原的鸡卵白蛋白偶联制备了抗β-conglycin的单克隆抗体。生成的单克隆抗体分别命名为6G4、3B7和5F10，经鉴定为igg1同型抗体，对β-甘氨酸具有高特异性。建立了基于6g4单克隆抗体的竞争性酶联免疫吸附法（ELISA）检测β-甘氨酸，ic50值为4.7ng/mL，检出限为2.0ng/mL。结果表明，基于6g4单克隆抗体的竞争性酶联免疫吸附试验重现性可靠。因此，制备的单克隆抗体6g4和建立的竞争性ELISA可为β-conglycin的灵敏检测和大豆β-conglycin相关食物过敏的研究提供有价值的工具。
Soybean β-conglycinin is one of the major food allergies for children and young animals. In order to detect immunoreactive β-conglycinin from soybean and soybean products, monoclonal antibodies against β-conglycinin were prepared using a conjugated chicken ovalbumin with a synthetic peptide that corresponded to one epitope sequence of β-conglycinin as the immunogen. The generated monoclonal antibodies, named as 6G4, 3B7, and 5F10, were identified as being of IgG1isotype and exhibited high specificity to β-conglycinin. A competitive enzyme-linked immunosorbent assay (ELISA) based on monoclonal antibody 6G4was established to determine β-conglycinin and showed an IC50value of 4.7ng/mL with a detection limit of 2.0ng/mL. The recovery tests of β-conglycinin indicated that the monoclonal antibody 6G4-based competitive ELISA gave reliable reproducibility. Therefore, the produced monoclonal antibody 6G4and the developed competitive ELISA could provide a valuable tool for sensitive determination of β-conglycinin and for future studies on food allergies related to soybean β-conglycinin.