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Development of ELISA for Detection of Mercury Based on Specific Monoclonal Antibodies Against Mercury-Chelate

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Abstract

Immunoassays for heavy metals offer an alternative approach to traditional techniques for detection of mercury. In this study, a mercury-chelate was prepared with 1-(4-aminobenzyl) ethylenediamine-N,N,N′,N′-tetraacetic acid (aminobenzyl-EDTA). The resulting complex was linked to keyhole limpet hemocyanin (KLH) or bovine serum albumin via the amino group and used as the immunizing antigen or detection antigen, respectively. BALB/c mice were immunized with KLH-aminobenzyl-EDTA-Hg and spleen cells from BALB/C mice were fused with Sp2/0 cells. One cell line (5F7) produced monoclonal antibodies with preferential selectivity and sensitivity for aminobenzyl-EDTA-Hg. This cell line had an affinity constant of 4.31 × 109 L/mol and its cross-reactivity (CR) with other metals was <2%. The antibody was used for competitive indirect ELISA (CI-ELISA) for Hg2+ measurements. The detection range was 0.087–790.4 μg/L and the lower limit of detection was 0.042 μg/L. The concentrations of mercury in environmental water samples obtained by CI-ELISA correlated well with graphite furnace atomic absorption spectrometry (GFAAS), and the mean recovery was 88.82% to 104.64%. These results indicate that this method could be used for monitoring mercury of water.

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Acknowledgment

This work was supported by the National “863” Project (2007AA10Z440) of China and the National Natural Science Foundation of China (Nos. 30972224, 31072178, and 30871897).

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Authors and Affiliations

  1. College of Animal Science and Veterinary Medicine, Jilin University, 5333 Xi’an Road, Changchun, 130062, China

    Yi Zhang, Xiaobing Li, Guowen Liu, Zhe Wang, Tao Kong, Jiajia Tang, Peng Zhag, Wei Yang, Dongna Li, Lei Liu, Guanghong Xie & Jianguo Wang

  2. College of Animal Science and Veterinary Medicine, Shenyang Agricultural University, Shenyang, 110161, China

    Yi Zhang

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  1. Yi Zhang
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  2. Xiaobing Li
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  3. Guowen Liu
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  5. Tao Kong
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  7. Peng Zhag
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  8. Wei Yang
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  10. Lei Liu
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  11. Guanghong Xie
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Corresponding authors

Correspondence to Xiaobing Li or Zhe Wang.

Additional information

Xiaobing LI and Guowen LIU contributed equally to this study.

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Zhang, Y., Li, X., Liu, G. et al. Development of ELISA for Detection of Mercury Based on Specific Monoclonal Antibodies Against Mercury-Chelate. Biol Trace Elem Res 144, 854–864 (2011). https://doi.org/10.1007/s12011-011-9031-z

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  • DOI: https://doi.org/10.1007/s12011-011-9031-z

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