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Preparation of hapten-specific monoclonal antibody for cadmium and its ELISA application to aqueous samples

Huan HE, Bo TANG, Cheng SUN, Shaogui YANG, Weijuan ZHENG, Zichun HUA

《环境科学与工程前沿（英文）》 2011年第5卷第3期页码 409-416 doi: 10.1007/s11783-011-0349-8

摘要： High-affinity and specific monoclonal antibodies against cadmium-ethylene diamine tetraacetic acid (EDTA) complex have been produced using the hybridoma technique. A hapten was synthesized and characterized by Fourier Transform Infrared Spectroscopy (FT-IR) and UV-Vis. Competitive enzyme-linked immunosorbent assay (ELISA) for quantitative detection of cadmium in aqueous sample was developed. The monoclonal antibody with high level of binding affinity for Cd-IEDTA-BSA and high specificity for soluble Cd-EDTA complex showed less than 0.99% cross-reactivity with other 11 metals. The limit of detection was 0.10 μg·L , and the effective linear range was 10 –10 μg·L . The intra- and inter-assay coefficient variations were 1.5%–6.3% and 3.2%–7.4%, respectively. The spike recovery in different water samples were between 98.5% and 110.3%. The detection limit of this assay was well below the allowable concentration of cadmium (3 μg·L ), and the working range was wider than that of other methods which showed the range of 2.19–86.38 and 0–10 μg·L . The competitive ELISA established in this paper was sensitive and accurate in the screening of cadmium in aqueous samples. The results will lay a solid foundation for construction of an immunoassay kit for cadmium.

关键词： cadmium hapten monoclonal antibody enzyme-linked immunosorbent assay (ELISA)

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Synthesis of haptens and production of antibodies to bisphenol A

Xiya ZHANG, Xiaoyun DONG, Sijun ZHAO, Yuebin KE, Kai WEN, Suxia ZHANG, Zhanhui WANG, Jianzhong SHEN

《农业科学与工程前沿（英文）》 2017年第4卷第3期页码 366-372 doi: 10.15302/J-FASE-2017132

摘要： Three immunizing haptens of bisphenol A (BPA), including two new haptens, were used to produce highly sensitive and specific polyclonal antibodies. The spacer arms of haptens for coupling to the protein carrier were located at different positions in BPA, and different length spacer arms were tested. Highly sensitive polyclonal antibodies were obtained and characterized using indirect competitive enzyme-linked immunosorbent assay (icELISA). Under optimized conditions, the half maximal inhibitory concentration (IC ) value of the best polyclonal antibody was 2.1 mg·L , based on coating heterogeneous antigens, and this optimal polyclonal antibody was highly sensitive toward BPA and displayed negligible cross-reactivity with bisphenol B and bisphenol E. A sensitive icELISA method utilizing the polyclonal antibody was developed for the determination of BPA in milk. In spiked samples (5, 10 and 20mg·L ), the recovery ranged from 80% to 102% with a coefficient of variation (CV) value below 15.8%. The limit of detection of icELISA was 1.95mg·L . These results indicate that the icELISA method is suitable for the detection of BPA in milk.

关键词： bisphenol A cross-reactivity hapten indirect competitive ELISA polyclonal antibody