Development of competitive ELISA for the detection of bovine serum albumin using single-chain variable fragments.
Development of competitive ELISA for the detection of bovine serum albumin using single-chain variable fragments.
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DOI：
10.1016/j.ab.2017.03.003
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发表时间：
2017-05
影响因子：
2.9
通讯作者：
Yuan Liu;Manman Lin;Xifeng Zhang;Xiaodan Hu;Jieru Lin;Jia Hao;Dan He;Xiao Zhang;Chongxin Xu;J. Zhong;Yajing Xie;Cunzheng Zhang;Xianjin Liu
中科院分区：
文献类型：
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作者：
Yuan Liu;Manman Lin;Xifeng Zhang;Xiaodan Hu;Jieru Lin;Jia Hao;Dan He;Xiao Zhang;Chongxin Xu;J. Zhong;Yajing Xie;Cunzheng Zhang;Xianjin Liu
在大肠杆菌HB2151中表达了可溶性抗牛血清白蛋白(BSA)单链可变区(ScFv)。表面等离子体共振分析测得单链抗体的抗原结合平衡解离常数为2.9g×10~(-1)−~8M。利用获得的抗体片段，建立了检测BSA的竞争酶联免疫吸附试验。添加牛奶样品的加标回收率在119.2%~87.8%之间，检出限(I_(10))和I_(50)分别为0.002和0.74μg/ml。该方法有可能取代基于传统抗体的酶联免疫吸附试验，用于检测牛奶或其他基质中的BSA浓度。
Soluble anti-bovine serum albumin (BSA) single-chain variable fragments (scFvs) were expressed inE. Coli.HB2151. The antigen-binding equilibrium dissociation constant of the scFvs was determined to be 2.9 × 10−8M by surface plasmon resonance analysis. A competitive ELISA for the detection of BSA was developed using the antibody fragment above. The limits of detection (I10) and I50were 0.002 and 0.74 μg/ml respectively, with a recovery between 87.8 and 119.2% in spiked milk samples. The assay has the potential to be used to detect concentration of BSA in milk or other matrix instead of the ELISA based on traditional antibodies.